Prevalence of Metagonimus metacercariae in sweetfish, Plecoglossus altivelis, from eastern and southern coastal areas in Korea

The present study was performed to determine the current infecion status of Metagonimus yokogawai metacercariae in sweetfish, Plecoglossus altivelis, collected from several streams in eastern and southern coastal areas of Korea. The sweetfish collected were artificially digested with pepsin-HCl solution and examined under a stereomicroscope in August and September, 2007. Out of 145 sweetfish collected from 10 streams in eastern coasts (Gangwon-do and Gye-ongsangbuk-do), 88 (60.7%) were infected with Metagonimus metacercariae. The average metacercarial density was 61 per infected fish. Among 141 sweetfish collected from 10 streams in southern coasts (Gyeongsangnam-do, Ulsan Metropolitan city, and Jeollanam-do), 140 (99.3%) were infected with Metagonimus metacercariae, and their average density was 949 per infected fish. The present study confirmed that M. yokogawai metacercariae are still prevalent in the sweetfish from several eastern and southern coastal localities. The prevalence and metacercarial density were much higher in the sweetfish from streams of southern coasts than in those of eastern coasts. Therefore, attention should be paid to this small fluke infection, and consumption of raw sweetfish naturally produced in these areas should be prohibited.     

Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai

We compared the DNA sequences of the genus Metagonimus: M. yokogawai, M. takahashii, and M. miyatai. We obtained 28S D1 ribosomal DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (mtCOI) fragments from the adult worms by PCR, that were cloned and sequenced. Phylogenetic relationships inferred from the nucleotide sequences of the 28S D1 rDNA and mtCOI gene. M. takahashii and M. yokogawai are placed in the same clade supported by DNA sequence and phylogenic tree analysis in 28S D1 rDNA and mtCOI gene region. The above findings tell us that M. takahashii is closer to M. yokogawai than to M. miyatai genetically. This phylogenetic data also support the nomination of M. miyatai as a separate species.     

Infection status of sweetfish from Kwangjung-stream and Namdae-stream in Yangyang-gun, Kangwon-do with the metacercariae of Metagonimus yokogawai

This study was performed to observe the infection status of sweetfish, caught from Kwangjung-stream and Namdae-stream in Yangyang-gun, Kangwon-do in August 1989, with the metacercariae of Metagonimus yokogawai. All of 28 sweetfish from Kwangjung-stream were infected with the metacercariae of M. yokogawai. The number of metacercariae in a fish ranged 89-521(224 on average), and the number of larvae/g of flesh was 12 approximately 55 (22 on average). On the other hand, no metacercariae were detected in the fish from Namdae-stream. It has been confirmed that the Kwangjung-stream in Yangyang-gun, Kangwon-do is one of the endemic foci of metagonimiasis in Korea.     

Structure of comblike teeth of the ayu sweetfish,Plecoglossus altivelis (Teleostei: Isospondyli): I. Denticles and tooth attachment

The structure and tooth attachment of the comblike teeth and denticles of the ayu sweetfish, Plecoglossus altivelis, were examined by light and scanning electron microscopy. The denticle is composed of a spoonlike crown with a spine pointed anteriorly, a triangular plate in the cervical region, and a root that curves laterally and tapers off to a point. The root apex is fused with a long thin pedicle that turns abruptly anteriad toward the jaw bone. Planes of the spine, the spoonlike crown, the triangle plate and the root of the denticle are varied, and the denticle is twisted in the region of the triangle plane. The superficial layer of the dentine is homogeneously calcified and is considered to be enameloid, because some of the inner dentinal epithelial cells in the tooth germ are columnar and possess cellular processes at their apical ends. The dentine is fibrous and fine dentinal tubules are visible in dentine treated with sodium hydroxide and observed by scanning electron microscopy. The upper half of the root is surrounded by a dense layer of collagen fibers running parallel to the tooth axis, and the lower half is encompassed by interlaced collagen fibers. The lower part of the root is open on its lingual side. The pedicle is a long rod which is homogeneously calcified and enmeshed by interlaced collagen fibers, and it curves mediad as it nears the jaw bone. The pedicles are interposed between a layer of gelatinous connective tissue and the jaw bone and terminate on the periosteum. Comparative aspects of ayu tooth morphology are discussed. © 1993 Wiley-Liss, Inc.     

Genetic characteristics of the amphidromous fish Ayu Plecoglossus altivelis altivelis (Osmeriformes: Plecoglossidae) on Yaku-shima Island in Japan,the southernmost population of the subspecies

Genetica - The Ayu (Plecoglossus altivelis altivelis) population on Yaku-shima Island, Kagoshima Prefecture, Japan represents the southernmost population of the subspecies and is considered to be...     

Development of free and canal neuromasts and their directions of maximum sensitivity in the larvae of ayu,Plecoglossus altivelis

Morphological changes in free neuromasts are reported from larvae of the Ayu,Plecoglossus altivelis. In newly-hatched larvae, free neuromasts were already recognizable in both the head and trunk. During larval growth, the number of free neuromasts increased, and the number of its sensory cells 2 days after hatching was constant. In the trunk, two types of free neuromasts, one with maximum sensitivity in the antero-posterior direction and the other with maximum sensitivity in the dorso-ventral direction, were observed. The former type predominated. In the head, free neuromasts were located around the eye and nose, their directions of maximum sensitivity forming lines tangential to concentric circles about the eye and nose. Distinct changes in free neuromasts occurred during the formation of the canal organ. The canal organ was first observed in the head region 64 days after hatching and in the trunk region 100 days after hatching. Concomitant with the formation of the canal organ, the profile of the cupulae of the free neuromasts changed from a flat bar to semispherical. Sensory cells in the canal neuromasts did not differ morphologically from those in the free neuromasts. It is considered that there is a close relationship between the sensitivity of the neuromast and the shape of the cupula, i.e., that the free neuromasts are adapted to slow water flow, as in lakes and the sea, while the neuromasts in the canal organ are adapted to rapid water flow.     

Occurrence and body size changes of drifting land-locked Ryukyu-ayu Plecoglossus altivelis ryukyuensis larvae in the San-numata River, Okinawa-jima Island, Japan

The occurrence and body size of drifting land-locked Ryukyu-ayu, Plecoglossus altivelis ryukyuensis, larvae were investigated in the San-numata River, Okinawa-jima Island from November 2000 to March 2001. The water temperature in the river fluctuated from 13.8 to 17.9°C during the sampling period. The drifting larvae occurred from mid-December to mid-March with an estimated spawning peak during late November and early December. The notochord length of the drifting larvae ranged from 4.9 to 6.7 mm (5.7 ± 0.3 mm, mean ± SD), and became longer as the spawning months progressed. These results imply that the land-locked population possesses the same maturation attributes as the original amphidromous population.     

Isolation and identification of non-chlorinated phenylbenzotriazole (non-ClPBTA)-type mutagens in the Ho River in Shizuoka Prefecture, Japan

We previously identified 2-[2-(acetylamino)-4-amino-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA) congeners as major mutagens in water concentrates from several rivers that flow in three different areas, i.e. Kyoto, Aichi, and Fukui Prefectures, in Japan. In synthesis studies, these PBTAs were shown to be formed from corresponding dinitrophenylazo dyes via non-chlorinated derivatives (non-ClPBTAs). However, only non-ClPBTA-1, i.e. 2-[2-(acetylamino)-4-[bis(2-methoxyethyl)amino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole, had been detected as a minor contaminant in the Nishitakase River in Kyoto. In this study, analysis of mutagens in water concentrate from the Ho River, which flows through an area with a textile dyeing industry in Shizuoka Prefecture, Japan, allowed the isolation of four compounds (I, II, III, and IV). These four mutagens were identified as 2-[2-(acetylamino)-4-[N-(2-cyanoethyl)ethylamino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-2), 2-[2-(acetylamino)-4-[(2-hydroxyethyl)amino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-3), 2-(2-acetylamino-4-amino-5-methoxyphenyl)-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-4), and 2-[2-(acetylamino)-4-(diethylamino)-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-7) by spectral data and co-chromatography using synthesized standards. Non-ClPBTA-3 and -7 were highly mutagenic in Salmonella typhimurium YG1024, inducing 159,000 and 178,000 revertants/microg, respectively, in the presence of S9 mix. Like PBTAs, non-ClPBTAs might have been produced from azo dyes during industrial processes in dyeing factories and released into rivers.     

Effects of photoperiod and temperature on testicular development in male ayu,Plecoglossus altivelis

Synopsis Growth, gonadosomatic index and plasma steroid profiles in male ayu,Plecoglossus altivelis, cultured under short/long photoperiods and cool/warm temperatures were determined. Juvenile males were assigned to each of four different photoperiod/temperature regimes (16 L/18°C, 16 L/24°C, 8 L/18°C and 8 L/24°C) at random. Fish were killed and examined bi-weekly over the following 16 weeks. Mean body weight in the 16 L/18°C treated fish was the highest among four treated groups. No significant differences between body weights of the 16 L/24°C, 8 L/18°C and 8 L/24°C treated groups were observed. Ayu in the 8 L/18°C treated group had the highest values of gonadosomatic index, plasma testosterone (T) and 17-hydroxyprogesterone (17-OH P). No significant differences of plasma E₂ were observed among the treated groups. In the 8L/18°C and 8L/24°C groups, peak levels of 17-OH P occurred after 12 and 14 weeks of treatment, respectively. No peak levels of plasma T and 17-OH P were observed in 16 L/18°C or 16 L/24°C treated ayu. Spermiation occurred only in ayu with 8 L/18°C treatment. The data suggest that testicular development in ayu is temperature and photoperiod dependent: short photoperiod and cool temperature favour gonadal development.     

Geographical distribution and cytological variation ofLemna paucicostata Hegelm. in Japan

More than 100 strains ofLemna paucicostata Hegelm. collected from various localities in Japan were classified into 4 types according to the criterion of Yukawa and Takimoto (1976). N-1 type strains are widely distributed in Hokkaido, Honshu, Shikoku and Kyushu, N-2 type strains only in the area on the Japan Sea side of north Japan, K type strain only at the campus of Kyoto University, and S type strains in southern Japan. Numbers of chromosomes in somatic cells of N-1, N-2, K and S type strains were 2n=66–84, 40, 50 and 40, respectively. In N-2 and S type strains, all chromosomes (20 bivalent chromosomes) were lined up on the equatorial plate at metaphase of the first meiotic division (MI). However, in N-1 type strains, several chromosomes, and in the K type strain, more chromosomes were away from the equatorial plate at MI. Pollen fertility was 60–70%, higher than 90%, 0% and higher than 90%, in N-1, N-2, K and S type strains, respectively.     

Seasonal dynamics of the fish parasite Neoplagioporus ayu (Digenea) in its definitive host, Plecoglossus altivelis, in the Chikugo River, Kyushu, Japan

Seasonal dynamics of the fish parasite Neoplagioporus ayu (Digenea; Opecoelidae; Plagioporinae) in its definitive host, the ayu Plecoglossus altivelis, in the Chikugo River, Kyushu, Japan, was examined. Natural ayu are amphidromous. In the upper Chikugo River, ayu fry raised in hatcheries are released into the river every spring because the migration of ayu is completely blocked by a dam that has no fish-pass channel. Ayu collected in April 2003 (within 1 month after release) harbored no N. ayu, but the prevalence of N. ayu rose to about 80% in May. The prevalence, abundance, and maturity of N. ayu were maintained at high levels from June to September. These finding indicate that the life cycle of N. ayu is completed in a freshwater area, although its intermediate hosts have not yet been identified despite the intense examination of invertebrates.     

Molecular cloning and characterization of rhodopsin in a teleost (Plecoglossus altivelis,Osmeridae)

Amplified fragments encoding exon-4 of opsin cDNAs were cloned from the retina of landlocked ayu (Plecoglossus altivelis), and sequenced. On the basis of the sequence homology to previously characterized fish visual pigments, one clone was identified as rod opsin (AYU-Rh), and two clones as green (AYU-G1, -G2), one as red (AYU-R) and two as ultraviolet (AYU-UV1, -UV2) cone opsins. The 335-amino acid sequence deduced from the full-length cDNA of AYU-Rh included residues highly conserved in vertebrate rhodopsins and showed the greatest degree (88%) of similarity with salmon rhodopsin. Southern blotting analysis indicated that ayu possess two rhodopsin genes, one encoding visual rhodopsin (AYU-Rh) and the other non-visual extra-ocular rhodopsin (AYU-ExoRh). RT-PCR experiments revealed that AYU-Rh was expressed in the retina and AYU-ExoRh in the pineal gland. In situ hybridization experiments showed that the mRNA of AYU-Rh was localized only in rod cells not in cone cells. Lake and river type landlocked ayu having different amounts of retinal and 3-hydroxyretinal in their retinas expressed a rhodopsin (AYU-Rh) of identical amino acid sequence.     

A mass occurrence of human infection with Diplogonoporus grandis (Cestoda: Diphyllobothriidae) in Shizuoka Prefecture, central Japan

Forty-six cases of human infection with Diplogonoporus grandis were found in Shizuoka Prefecture on the Pacific coast of Central Japan in 1996. The cases were predominantly elderly male patients over 50 years of age. Although all cases were reported from May to September of the year, most of them were diagnosed in June and July. We suspected that the transmission was due to the consumption of raw juvenile Japanese anchovy (Engraulis japonicus), which are seasonally caught in the spring months off the Pacific coast of the Prefecture. In almost a hundred years after its discovery in 1894, there had been more than 180 cases of human diplogonoporiasis recorded in Japan. The high incidence within a relatively short time frame of our investigation is regarded unusual even in this country.     

Lectin-reactive components of the microsporidian Glugea plecoglossi and their relation to spore phagocytosis by head kidney macrophages of ayu Plecoglossus altivelis.

We investigated the reactivity of lectins to spores of Glugea plecoglossi from ayu Plecoglossus altivelis. Smear preparations of purified spores were treated with 8 kinds of lectins. Lectin blots were used to detect glycoproteins of spore lysates. In addition, lectin-treated spores were applied to head kidney macrophages of ayu, and the percentage of phagocytosis (PP) was calculated and compared with the control. Two lectins (ConA, WGA) reacted with the surface of the spores, and a major band (55 kDa) and some minor bands were visualized on blots after treatment with these. PP was decreased after ConA treatment. From these results, we suggest that G. plecoglossi spores can be phagocytized by ayu head kidney macrophages via ConA-reactive glycoprotein-mediated recognition.     

Pattern of synaptic connections in the pineal organ of the ayu,Plecoglossus altivelis (Teleostei)

Summary Synaptic connections were studied by means of electron microscopy in the sensory pineal organ of the ayu, Plecoglossus altivelis, a highly photosensitive teleost species. Three types of specific contacts were observed in the pineal end-vesicle: 1) symmetrically organized gap junctions between the basal processes of adjacent photoreceptor cells; 2) sensory synapses endowed with synaptic ribbons, formed by basal processes of photoreceptor cells and dendrites of pineal neurons; 3) conventional synapses between pineal neurons, containing both clear and dense-core vesicles at the presynaptic site. Based on these findings, the following interpretations are given: (i) The gap junctions may be involved in an enhancement of electric communication and signal encoding between pineal photoreceptor cells. (ii) The sensory synapses transmit photic signals from the photoreceptor cells to pineal nerve cells. (iii) The conventional synapses are assumed to be involved in a lateral interaction and/or summation of information in the sensory pineal organ. A concept of synaptic relationships among the sensory and neuronal elements in the pineal organ of the ayu is presented.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany     

Oral ontogeny of the Ayu, Plecoglossus altivelis and comparisons with the jaws of other salmoniform fishes

The Japanese Ayu, Plecoglossus altivelis Temminck & Schlegel is the sole representative of the salmoniform family Plecoglossidae. The Ayu is remarkable for its dentition which in adults comprises groups of diagonally arranged comb-like teeth in the outer tissue of the jaws. In juveniles (below 63 mm SL) the teeth are attached normally to the jaws. The transition of tooth form is correlated with a switch from zooplanktivory to algal or aufwuchs grazing. The present study follows the development of the teeth, jaws, oral cavity ethmoid and suspensorial elements in specimens ranging in size from 41–70 mm SL. The possible mode of function of the adult dentition is discussed. Comparisons are made with the jaws of other salmoniform fishes and a suite of supposed apomorphic characters are identified which are also shared with certain genera of the family Osmeridae, thus supporting the ideas of others that the Osmeridae is a paraphyletic assemblage.     

Utilization of α-starch in ayu, Plecoglossus altivelis, relating to growth and body composition

To examine the possibility of dietary α‐starch in reducing feed costs in a practical diet, α‐starch was supplemented at 10, 20, 30 and 40% in a composed diet having the same protein level. The four diets were fed to ayu, Plecoglossus altivelis (initial weight 9.1 g) for 43 days. Growth and feed efficiency increased with the supplement, with values highest in the 30–40%α‐starch diet. The level of dietary α‐starch did not affect the proximate muscle composition; although the hepatosomatic index was not affected, liver glycogen increased with increasing dietary α‐starch. The dietary α‐starch did not influence evacuation time from the gut, and was well digested through passage in the gut, mainly between the stomach and the anterior part of the intestine. Ayu have an ability to adapt their metabolism to high dietary α‐starch, and can digest 40% or more in a composed diet. Although the muscle lipid content did not change, the fatty acid composition was influenced by dietary starch. With the elevation of dietary starch, a decrease of C18:2n‐6 and an increase of C22:6n‐3 occurred. These results indicate that at least 40%α‐starch can be used in practical diets for ayu.