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PubMed1998

Life history of Echinoparyphium recurvatum (Trematoda: Echinostomatidae) in Korea.

The present study was performed to observe characteristics of the life history of Echinoparyphium recurvatum under both natural and laboratory conditions in Korea. A batch of Radix auricularia coreana was collected from Sunamchon, one of the stream of West Naktonggang (River), in Kangso-gu, Pusan during August and September 1992. Out of 106 snails examined by crushing, 52 (49.0%) were infected with larval E. recurvatum, i.e. rediae, cercariae and metacercariae. Cercariae naturally shed from snails encysted in the snails of same species and loaches, but not in mud-snails. Adult worms were detected from chicks and ducks experimentally infected with metacercariae, but not from rats and mice. The average recovery rate of adults from chicks was 13.1%. Rediae were sac-like, 2.437 x 0.317 mm in average size, with a muscular pharynx and a brownish cecum which reached the anterior half of the body. Cercariae consisted of a spindle-shaped body (0.262 x 0.129 mm in average) and a rod-like tail (0.528 x 0.056 mm in average). In the cercarial body, 45 collar spines were observed on the head crown, and double rows of excretory ducts with fine granules were laterally arranged between the pharynx and the ventral sucker. Metacercariae were spherical, 0.144 x 0.142 mm in average size, with thick hyaline outer and thin elastic inner walls, and many excretory granules. Adults were slender and more attenuated in the anterior end, 2.760 x 0.550 mm in average size, and had 45 collar spines including four end group spines on both ventral corners. From the above results, it was confirmed that R. auricularia coreana plays a pivotal role in the life cycle of E. recurvatum as the first and/or second intermediate hosts in Korea.

Animals↗
PubMed2000

Isolation of a lethal rhabdovirus from the cultured Chinese sucker Myxocyprinus asiaticus.

A rhabdovirus was found to be associated with a lethal hemorrhagic disease in the cultured Chinese sucker Myxocyprinus asiaticus Bleeker. The rhabdovirus was amplified and isolated from the infected GCO (grass carp ovary) cells. In ultrathin sections of liver cells from the diseased fish, the virus particles exhibited the characteristic bacilliform morphology, and budded through vesicle membranes of the infected cells. The isolated rhabdovirus particles were found to have a bacilliform morphology with 2 rounded ends rather than a typical flat base. The virus particles were measured and ranged in size from 150 to 200 nm in length and 50 to 60 nm in diameter. Most other characteristics, including their size, extensive virus infectivity to fish cell lines, strong cytopathogenic effects, stability at high temperatures, vesicle formation in infected cells, structure protein electrophoretic patterns and the presence of an RNA genome, very closely resembled those of other fish rhabdoviruses. At present it is not known if this is a novel virus species or if it is an isolate of a known fish rhabdovirus. Until a confirmed identification can be made, we will temporarily refer to this virus as Chinese sucker rhabdovirus (CSRV).

Animals↗
PubMed2004

Genetic relationship among gill-infecting Myxobolus species (Myxosporea) of cyprinids: molecular evidence of importance of tissue-specificity.

The importance of tissue-specificity was studied in Myxobolus spp. infecting the gills of 7 cyprinid species. The 18S rDNA of 10 Myxobolus species was amplified by optimised nested-PCR, resulting in approximately 1600 bp PCR products. Phylogenetic trees generated by distance matrix and parsimony analyses revealed 4 main groups. Muscle-infecting species all belonged to the same group, while members of 2 gill-infecting groups were clearly distinguishable on the basis of tissue-specificity, and were also recognisable by differences in spore morphology. On the basis of tissue tropism, phylogenetic relationships among the species examined indicate that genetic separation is a more ancient evolutionary feature than host-specificity.

Animals↗
PubMed1989

Metacercariae of Diplostomum spathaceum in the eyes of fishes from Yellowstone Lake, Wyoming.

Fish were collected from Yellowstone Lake, Wyoming (USA). Metacercariae of Diplostomum spathaceum was found in the lens of 11 of 12 longnose suckers (Catostomus catostomus). The mean number of metacercariae per sucker was 59 and the average age of the fish was 11.6 yr. There was no correlation between age and intensity of parasites (r = 0.24). Of 10 cutthroat trout (Salmo clarki) examined, there were metacercariae present in six. The metacercariae were found outside of the lens tissue in the trout; they occurred in the vitreous humor and the retina. These may be a different species from those found in the suckers.

Animals↗
PubMed1999

Genomic organization and sequence of the mud loach (Misgurnus mizolepis) growth hormone gene: a comparative analysis of teleost growth hormone genes.

The mud loach (Misgurnus mizolepis) growth hormone (GH) gene was cloned and a comparative analysis on its genomic organization was performed. Based on Southern analysis using various kinds of restriction endonucleases, the GH gene proved to exist as a single-copy gene in the mud loach. The complete nucleotide sequences of a 5.1 kb SacI/EcoRI genomic fragment containing the mud loach GH gene and its 5' flanking sequences as well as a mud loach GH cDNA obtained by rapid amplification of a reverse transcriptase-PCR have been determined. The GH gene spans 2.0 kb from the start codon to the polyadenylation signal, and contains five exons and four introns similar to those of carps and mammals. The evolutionary relation of the mud loach GH gene, inferred by comparative analyses of gene structures and sequences in each exon and intron of representative teleost GH genes, reflects the major phylogenetic groupings of teleost.

Amino Acid Sequence↗
PubMed2000

[The chromosome mapping of PdSox8 and PdSox9 in two kinds of loaches].

With the Dig-labelled PdSox8 and PdSox9 as probes, the chromosome mapping of Sox8 and Sox9 in Paramisgurnus dabryanus and Misgurnus anguillicaudatus were analyzed by chromosome in situ hybridization technique. In P. dabryanus, the PdSox8 and PdSox9 were successfully localized in the No. 4 and No. 2 telocentric chromosome respectively, the relative distance to centromere is 40.2% and 67.5%; In M. anguillicaudatus, they were localized in the No. 9 and No. 6 telocentric chromosome respectively, the relative distance to centromere is 58.3% and 30.8%.

Animals↗
PubMed2000

Predation efficacy of the fish muddy loach, Misgurnus mizolepis, against Aedes and Culex mosquitoes in laboratory and small rice plots.

Assessments of the biological control potential of muddy loaches, Misgurnus mizolepis, were conducted against Aedes togoi, Culex pipiens pallens, and Culex inatomii larvae both in the laboratory and in small rice plots in Pusan, Korea, from June to September 1997. Medium-sized fish consumed almost all of 500 3rd-stage larvae of Cx. pipiens pallens offered daily in the laboratory. Predation by the fish at release rates of 1, 2, 4, 8, and 16 fish per plot resulted in 1,004, 1,197, 1,198, 1,200, and 1,200 larvae of Ae. togoi consumed in 12 h, respectively, when each 1,200 3rd-stage larvae were given in the 1-m2 small rice plots. The average numbers of larvae eaten after introduction of the fish were 1,121.8 larvae in 24 h and 1,195.8 larvae in 36 h. Muddy loaches showed slightly higher predation rates on Cx. pipiens pallens and Culex inatomii larvae than on larvae of Ae. togoi. Therefore, M. mizolepis might be an efficient biological control agent against mosquito species in rice fields.

Aedes↗
PubMed2000

Local changes in gap junctional permeability accompany regionalization of the mesoderm in early fish (loach, Misgurnus fossilis) embryos.

The marginal area of early loach (Misgurnus fossilis) embryos in the proximity of the yolk cell (YC), the zone of early activation of mesoderm-specific genes, was revealed to be characterized by regional changes in the permeability and selectivity of gap junctional (GJ) channels. Comparison of the efficiency of GJ transfer of negatively (Lucifer Yellow, LY and fluorescein, F) and positively (4',6-diamidino-2-phenylindole, DAPI) charged dyes showed that, at the early gastrula stage, GJs formed between blastomeres of the presumptive mesoderm layer and the YC acquired a higher permeability for LY and F and became virtually impenetrable for DAPI over most of the germ ring area, except for its dorsal side. GJs in the dorsal-anterior region were permeable for both LY and DAPI. The dorsal-posterior area was labelled with neither of the dyes. Observations of the fate of embryonic shield blastomeres stained with both LY and DAPI showed that their progenies were confined to the head mesoderm, whereas the paraxial somite mesoderm and the ventral mesoderm contained the cells labelled only with LY or F. The presumptive notochord region was not labelled at all. These data support the hypothesis that GJs between the embryonic cells may be involved in the spatial organization of development due to their ability to transfer different regulatory factors in different directions, filtering molecules in accordance with the electric charge, conformation and size.

Animals↗
PubMedDate not supplied

[Spermatozoa of the loach Misgurnus fossilis as a test system for identification of new centromere proteins].

We studied the possibility of using the spermatozoa of the loach Misgurnus fossilis L. for identification of centrosome proteins. It has been shown that the centrosome of the loach spermatozoa consists of a pair of centrioles of the standard structure and contains the marker protein gamma-tubulin, cytoplasmic microtubules branch out from it, and it does not contain any additional structures characteristic of the centrosomes of spermatozoa of many other fishes. A preparation enriched with intact centrosomes has been obtained from the loach spermatozoa. These centrosomes contained gamma-tubulin although they lost their ability to induce polymerization of microtubules. The preparation of loach centrosomes was successfully used to obtain a set of monoclonal antibodies against the mammalian centrosome. A new protein kinase LOSTEK was identified with the help of one of these monoclonal antibodies, SN2-3D2, which was localized in the centrosome and on then microtubules in both loach spermatozoa and cultured mammalian cells. Hence, the loach spermatozoa are a promising object for identification of new proteins of the mammalian centrosome.

Animals↗
PubMedDate not supplied

[Regularity of biochemical differentiation in ontogenesis].

Here we review original data on biochemical differentiation mechanisms. The patterns of the preprogrammed metabolism of the embryo are studied by comparing biochemical mechanisms in undifferentiated embryonic tissues in the course of differentiation and during the terminal establishment of a specialized tissue. This review is composed of three main sections corresponding to the specificity of biochemical differentiation. The first section covers the material on time-related isozyme pattern during skeletal muscle differentiation in a loach (a teleostean fish). The second section presents the data on "incorporated activation", i.e., the appearance of a functional relationship between contractile proteins biosynthesis and the systems of glycogenolysis and glycogen synthesis during the development of chick embryo muscle. The third section covers the data on liver differentiation, the formation of the glycogen organelle with embedded enzymes of glycogen synthesis, and degradation during the development of chick and rat embryos.

Animals↗
PubMed2001

[Intracellular localization of calcium in loach embryonic cells at the early gastrula stage].

Intracellular localization of calcium-accumulated structures in the loach embryo cells at the state of early gastrulation was determined by electron microscope histochemical pyroantimonate method. Calcium-precipitate was observed in the nucleus (external and inner membranes, chromatin and nucleolus), and in the cytoplasm (ER, mitochondria and submembrane cortical layer). Our present data show that such cell organelles as nuclear envelope, ER, and mitochondria serve as the basic cellular stores for calcium, confirming the existence of calcium signal system both in the nucleus and in the cytosol.

Animals↗
PubMed2001

[Transgenic loach produced by using sperm cells mediated by high molecules].

The pCH110 plasmid DNA which has a marker gene LacZ controlled by a SV40 promoter was mixed with a kind of high molecule of dendrimers(cyclic core dendrimer) and then formed a DNA/dendrimer complex. Loach sperms were incubated with the DNA/dendrimer complex. Then in situ hybridization was used to detect whether the sperm cells captured the foreign DNA or not. Results show that the number of sperm capturing foreign DNA via dendrimers as vectors was obviously increased. The sperms were mixed with eggs for in vitro fertilization. PCR was used to detect the transgenic loach fries. Obvious expression of LacZ gene in the head of transgenic loach were observed by analysis through LacZ histochemical staining of embryos or fries.

Animals↗
PubMed2001

[Study on immunonodulating activities of enzymolyzed loach protein].

The immunomodulating activities of enzymolyzed loach protein in mice were investigated by regular methods for immunology evaluation. The doses of enzymolyzed loach protein were 0.1, 0.2, 0.4 g.kg-1 bw, which were 5, 10, 20 times respectively as large as the recommended daily intake(0.02 g.kg-1 bw). After being successively fed with enzymolyzed loach protein at given doses for 30 days by ig respectively, the mice were detected by immunomodulation tests. The results indicated that enzymolyzed loach protein had significant modulating effects on all of test indexes and suggested it had apparent immunomodulating activities in mice. Based on the results described above, we speculate that the enzymolyzed loach protein may have immunomodulating functions and can enhance the immunity of body.

Adjuvants, Immunologic↗
PubMed2002

Motility of minute intestinal fluke, Haplorchinae spp, metacercariae in fish dishes prepared by different uncooked methods.

CONTEXT: Fish-borne trematode is a worldwide problem, with the number of people infected by liver flukes alone estimated at 21 million. In addition to the major liver flukes, Opisthorchis viverrini and Clonorchis sinensis, several types of intestinal flukes also use cyprinoid fish as the intermediate host. Traditional ingestion of undercooked and uncooked fish preparations is a major factor in pathogen acquisition. OBJECTIVE: To examine the motility of Haplorchinae spp metacercariae in matched minnows using a number of preparation methods that mimic traditional practices. DESIGN: Several freshly killed freshwater fish, Cyclocheilichthys armatus, were purchased from the local market and sent to the authors' laboratory to be examined under a stereoscopic microscope for active Haplorchinae spp metacercariae. A total of 10 fish were infected with many active metacercariae and used in the preparation of uncooked fish dishes. Five different raw fish dishes that mimic traditional meals (2 fishes/dish) were prepared using the following methods: 1) left to dry at room temperature; 2) frozen at -20 Celsius; 3) refrigerated at 4 Celsius; 4) marinated in saline (5% sodium chloride solution); and 5) marinated in 5% acetic acid solution to mimic traditional vinegar. The motility of the metacercariae in each of these dishes was examined under a stereoscopic microscope. Motility was estimated as the level of activity or degeneration of the metacercariae at the start of the experiment and then every 30 minutes until all the parasites appeared degenerate (and therefore, presumably not viable). RESULTS: Degeneration of the parasites was slowed by cooling: degeneration of all metacercariae took approximately 5 hours in the refrigerated or frozen fish, compared with 3 hours in all other dishes left at room temperature. CONCLUSION: The study substantiates the epidemiologic data demonstrating transmission of these types of pathogens despite freezing, pickling, and salting. Although the tradition of eating uncooked fish is deeply rooted in Thailand, constant efforts need to be made to discourage this practice.

Acetic Acid↗
PubMed2002

Molecular cloning and characterization of ARS elements from the mud loach (Misgurnus mizolepis).

Autonomously replicating sequences (ARSs) are thought to occur within, or adjacent to, the matrix attachment regions (MARs). To identify fish ARSs, MARs of the mud loach fish were obtained from nuclear matrices using a modified LIS method. These DNA fragments were screened for their ability to act as ARSs by being cloned into the ARS cloning vector, pURY19, and transformed into Saccharomyces cerevisiae. Sixteen ARSs were isolated, most of which were more efficient in transformation than the positive control vector, pURY19-2 microm, which contained the 2 microm circle origin of yeast. In particular, one clone, pURY19-ARS223, was 18 times more efficient in back-transforming E. coli than the positive control vector. Therefore, ARS223, which has strong ARS activity in yeast, could be a good candidate for inclusion in expression vehicles that are used to transfect fish cell lines or embryos. A DNA sequence analysis showed that the essential ARS elements contain potential ARS consensus sequences, and are predicted to have hairpin loop structures, or curved or kinked DNA. In addition, the MAR-Finder program suggested that ARSs also contain MAR motifs. These include AT tracts, ORI patterns, kinked DNA, ATC tracts, and Topoisomerase II consensus sequences. The in vitro matrix binding assay confirmed that all of the cloned ARSs could associate with the nuclear matrix. This indicates that ARSs elements may be located in or near the MARs. This is the first study that has identified and characterized ARSs in fish.

Animals↗
PubMed2002

Effect of Misgurnus anguillicaudatus polysaccharide on immune responses of splenocytes in mice.

AIM: To investigate the effect of Misgurnus anguillicaudatus polysaccharides (MAP) on immune responses of splenocytes in mice. METHODS: T lymphocyte proliferation (TLP) was measured by [3H]thymidine incorporation assay. Cytotoxic T lymphocyte (CTL) cytotoxicity and natural killer (NK) activity were determined by release of radioactive chromium [51Cr] from pre-labeled target cells. RESULTS: MAP 5 and 10 mg . kg-1 . d-1, ip for 7 d, could increase TLP, enhance the cytotoxicity of CTL and NK cells, and antagonize the effect of concanavalin A (ConA) on TLP suppressed by cyclophosphamide (CP). Inhibitory rates of CTL cytotoxicity were decreased from 51.4 % in CP control mice to 18.2 % and 35.1 % in MAP-treated CP mice, respectively. Furthermore, administration of MAP 10 and 20 mg . kg-1 . d-1, ip for 7 d, restored the reduced NK cell cytotoxicity caused by CP administration in mice. CONCLUSION: MAP has protective effect on augmenting T-cell-mediated immunity and NK activity in normal and CP-treated mice.

Adjuvants, Immunologic↗
PubMedDate not supplied

[Statistical and frequency-amplitude characteristics of ultra weak emissions of the loach eggs and embryos under the normal conditions and during their optic interactions. I. Characteristics of ultra weak emission in normal development and the optic role of egg envelope].

Ultraweak emissions of groups comprising several dozens of unfertilized and fertilized loach eggs, embryos, larvae, and their egg envelopes were measured on a photomultiplier tube. The envelopes absorbed the light from external sources but readily gave it back in the absence of embryos. We carried out statistical and frequency-amplitude analyses of ultraweak emissions and studied the autocorrelation structure of their frequency spectra. The frequencies of signals with different intensity underwent regular changes during development. Cascades of short-term (< or = 1 ms) flashes timed (during cleavage) to furrowing were a characteristic element of ultraweak emission. The Fourier spectra of developing embryos had pronounced frequency-amplitude peaks and higher, than in unfertilized eggs and inanimate samples, mutual correlation during successive time intervals. Stage-specific translational symmetry of the frequency spectra of ultraweak emissions was demonstrated, which suggests the presence in groups of embryos of a coordinated system of harmonic oscillators. The latter underwent regular changes during development. The measurement of ultraweak emissions represents a unique non-invasive method of analysis of these oscillators.

Animals↗