Characterization of the goldfish fecal microflora by the fluorescent in situ hybridization method

ABSTRACT:   Bacterial populations in goldfish feces were characterized by the fluorescent in situ hybridization (FISH) method. A total of nine different group-specific rRNA-targeted oligonucleotide probes were used. Approximately half of the microbial cells (57.8 ± 16.7%) were detected with a probe EUB338 and found to be bacteria. The microbial cells in 33–35 of the 35 samples from five specimens strongly hybridized with probes ALF1b, BET42a and GAM42a, suggesting that goldfish intestinal bacteria are mainly composed of α, β and γ-subclasses of Proteobacteria. The fact that a probe AER66 reacted with 25.6 ± 14.2% of the total microbial cells in all 35 samples, demonstrated that genus Aeromonas was the dominant species in the goldfish intestines. Genus Bacteroides including Bacteroides type A detected with a probe BAC303 was observed in 15 of 35 samples while other taxonomic groups determined with HGC69a, CF39a and P72 were detected in 6–11 of 35 samples. These results strongly suggest that Bacteroides shows the greatest daily fluctuation and interindividual variation in the intestines of goldfish. Moreover, the FISH method was proven to be useful for rapid enumeration of taxonomic groups of fish intestinal bacteria.     

Gonadotropin-releasing hormone and gonadotropin in goldfish and masu salmon

Reproductive activities in vertebrates are regulated by an endocrine system, consisting of the brain-pituitary-gonad axis. In teleosts, gonadotropin-releasing hormone (GnRH) in the brain stimulates gonadotropin (GTH) release in the pituitary gland, but because of lack of the portal vessel, it is not known when and how much GnRH is released for the regulation of GTH release. There are multiple molecular types of GnRH in teleosts and several distinct populations of GnRH neurons in the brain. However, we do not know which types and populations of GnRH neurons regulate reproductive activities. Here we summarize our recent studies on GnRH and GTH in masu salmon Oncorhynchus masou and goldfish Carassius auratus. Immunocytochemistry showed the location and molecular types of GnRH neurons. Salmon (sGnRH) and chicken-II GnRH (cGnRH-II) neuronal fibers were widely distributed in the brain of both masu salmon and goldfish. Only sGnRH fibers were observed in the pituitary of masu salmon, whereas both sGnRH and cGnRH-II fibers were observed in the goldfish pituitary, indicating that species specific GnRH profiles are involved in the regulation of pituitary function in teleosts. A series of experiments in masu salmon and goldfish suggest that among GnRH neuron populations GnRH neurons in the ventral telencephalon and the hypothalamus regulate GTH release, and that GnRH of the terminal nerve origin is not essential to gonadal maturation and ovulation. The biological function of other GnRH neurons remains unkown. Two GTHs appear to be characteristic of teleost; however, regulation of reproduction by these GTHs is a question that remains to be elucidated. In salmonid species, it is proposed that GTH I stimulates early gonadal development, whereas GTH II acts in later stages. When GTH expression was examined in goldfish, both GTH I and II mRNA levels in the pituitary gland showed increases in accordance with gonadal development, unlike the sequential expression of GTH subunits in salmonids. The expression of these GTH subunit mRNAs were affected by water temperature, starvation, and steroid hormones in goldfish, but in what manner these two GTHs regulate gonadal development remains to be clarified.     

四种脊椎动物小脑皮层的显微结构对比

分别就鲫鱼、黑斑蛙、石龙子、小自鼠四种脊椎动物小脑皮质的显微结构做了初步的观察和对比，发现在这四种动物的小脑皮质中，主要含有同样的三种神经细胞：分子层细胞、蒲肯野细胞、颗粒细胞。     

Color vision, spectral sensitivity, accommodation, and visual acuity in juvenile masu salmon Oncorhynchus masou masou

ABSTRACT:   Color vision, spectral sensitivity, accommodation, and visual acuity were examined in juvenile masu salmon Oncorhynchus masou masou to obtain fundamental information about the visual system. Two types of S-potentials were recorded from 415 horizontal cells in isolated retinas from 34 cultured freshwater masu salmon (114–219 mm standard length, SL). Although horizontal cells recording S-potentials were not identified, the horizontal cells were confirmed because their responses were maintained while the stimulus remained. The variety of chromaticity (C)-type S-potentials indicated well-developed color vision. The analysis of the luminosity (L)-type S-potentials indicated that the peak spectral sensitivity was at 522 nm. S-potentials were also recorded in response to ultraviolet light. The direction and extent of lens movement induced by electrical stimulation was measured in 12 cultured masu salmon (99.0–142.5 mm SL). The results indicated that the visual axis was upward and forward, and that the range of accommodation was from 0.79 × SL in front of the eye to infinity. In histological analysis of the retinas of five wild smolts (100–118 mm SL), the maximum cone densities (276–345 cones/0.01 mm²) were detected in the ventral to temporal regions. The visual acuities assessed by histological methods were 0.069–0.075.     

A new method for recording activities of spinal motoneurons during fictive swimming in paralyzed teleost fish

ABSTRACT: A new method for monitoring spinal motoneuron activities during fictive swimming in teleost fish was developed. Enamel-insulated copper wire electrodes were implanted in the trunk muscle of goldfish or carp. For each freely moving fish, an electromyogram (EMG) was recorded using the electrodes. In those fish paralyzed with curare, using the same electrode set, bursts of electrical activities consisted of spikes of smaller amplitude and of shorter duration compared with those recorded by EMG. Simultaneous recording of the extracellular activity and intracellular recording from the muscle revealed that the bursts of spikes recorded in the paralyzed fish were motor nerve impulses innervating the muscle and are considered to be fictive swimming activity. The method developed in the present study provides a useful tool with which to investigate neuronal mechanisms underlying swimming activity in teleost fish.     

Colouring ornamental fish (Cyprinus carpio and Carassius auratus) with microalgal biomass

Koi carp and goldfish value increases with intensity of skin colour, which is an important quality criterion. Fish cannot fully synthesize their own carotenoid colourings and these must therefore be included in their diet. Two trials were undertaken to investigate skin colour enhancement in ornamental species (i.e. three chromatic varieties of koi carp (Cyprinus carpio), namely Kawari (red), Showa (black and red) and Bekko (black and white) and goldfish (Carassius auratus)) by feeding a dietary carotenoid supplement of freshwater microalgal biomass [Chlorella vulgaris, Haematococcus pluvialis, and also the cyanobacterium Arthrospira maxima (Spirulina)], using a diet containing synthetic astaxanthin and a control diet with no colouring added for comparison. In the first trial, five homogeneous duplicate groups of 25 juvenile koi carp (C. carpio) (initial mean body weight 24.6 ± 0.7 g) were fed, for 10 weeks, one of the four diets containing 80 mg colouring/kg diet. In the second trial, this procedure was repeated for five homogeneous duplicate groups of 25 goldfish (C. auratus) (initial mean body weight of 0.9 ± 0.1 g). Initial and final samples of skin along the dorsal fin were withdrawn, from five fish per group, for subsequent analysis of total carotenoid content (spectrophotometric analysis), and red hue (colorimetric analysis, CIE (1976) L* a* b* colour system). Growth and feed efficiency were not significantly different between groups administered by the various dietary treatments. In both trials, dietary carotenoid supplementation increased total skin carotenoid content. The more efficient colouring for koi carps was found to be C. vulgaris biomass, providing both maximum total carotenoid deposition and red hue for the three chromatic koi carp varieties studied, and particularly for the kawari variety. For goldfish the best colouring obtained, as ascertained by total carotenoid content, was also achieved using C. vulgaris biomass, and red hue was maximum when using H. pluvialis biomass.     

几种物质对金鱼的诱食活性

本文采用迷宫装置、通过液体实验的方法,研究了二甲亚砜和5种市售香精对金鱼的诱食活性。结果表明:二甲亚砜、葡萄香精和菠萝香精对金鱼有显著的诱食活性,而杨梅香精、荔枝香精和桔子香精则无诱食活性。     

Structure and function of the central nervous system of domestic animals

The structure of the central nervous system is described in domestic animals. Differences in contrast to the human CNS and the functions of the nervous centers are determined. The neurons which are situated on the surface of the telencephalon and cerebellum are arranged in a laminated way. In other areas the neurons are clustered and form nervous nuclei. The latter are built up normally by large, middle large and small neurons. Lightmicroscopically they show a large nucleus poor in chromatin and a prominent nucleolus. At the ultrastructural level the pericaryon is rich in Nissl-bodies, formed by a prominent tubulus-system of granular endoplasmic reticulum. The surface of the cell bodies is covered by three different types of synaptic junctions. In the neuropil there are five different types of synapses. The ultrastructure of these synapses, its functions and transmitter substances, which are located inside the synaptic vesicles, are described.     

Influence of lupin and rapeseed meals on the integrity of digestive tract and organs in gilthead seabream (Sparus aurata L.) and goldfish (Carassius auratus L.) juveniles

Two rearing trials were conducted to evaluate the effects of dietary lupin (LM) and rapeseed (RM) meals in gilthead seabream (Sparus aurata L.) and goldfish (Carassius auratus L.) juveniles. Each plant meal was incorporated at the rate of 200 g kg^?1 in two distinct diets, which were compared with a fishmeal‐based diet as control. After 1 month, the two plant diets did not influence the whole body growth, but the digestive systems were affected. The splenosomatic index was reduced with two plant meals in goldfish and with RM in seabream. The hepatosomatic index was only reduced in LM‐fed seabream. Cellular characteristics were also affected. The largest liver cells were observed in RM‐fed goldfish suggesting changes in metabolic function. The LM and RM diets stimulated in seabream, especially the reaction in haematopoietic tissues with the proliferation of melano‐macrophages centres, and a tendency for elongated villus height in the anterior intestine thus that possibly compensated for a reduction in digestive function. Such adaptive structural modifications and the absence of degenerative signs allowed concluding that the integrity of the digestive system was maintained in fish fed plant meals.     

Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms

Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus , caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10⁷ copies μg⁻¹ DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10⁶–10⁸ copies of CyHV-2 μg⁻¹ DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22–10 °C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (±SE) of 7.3 ± 11 to 394 ± 55 μg⁻¹ DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease.