The appearance pattern of tooth germs in the round crucian carp,Carassius auratus grandoculis

Cyprinid fishes generally replace their teeth alternately and cephalad. The larvae ofCarassius auratus grandoculis also replace their teeth alternately and cephalad, in a pattern of 4-2-3-1-. However, adults ofCarassius species replace their teeth from anterior to posterior, in a pattern of 1-2-3-4-1-. So I analyzed the appearance pattern of tooth germs in larvae and juveniles inCarassius auratus grandoculis. At stage 5 of the post-larval period, developmental difference is made between both sides. In the pharyngeal dentition on one side developing poorly, the anterior tooth on the fifth replacement wave, tooth₄[An2] appeared later than the central teeth on following replacement wave, tooth₅[Pol]. Moreover, the anterior tooth on the seventh replacement wave, tooth₆[An2], appeared later than the central teeth on the following replacement wave, tooth₇[Pol], on both sides. The reverse of tooth germ appearance between anterior teeth and central teeth makes a change of replacement pattern from 4-2-3-1-4- to 1-2-3-4-1-. The change of replacement pattern is caused by the confusion of tooth germs of anterior teeth on both sides.Mylopharyngodon piceus andCyprinus carpio make a change of replacement patterns in the early juvenile period, too. This change of replacement pattern may be a specialized character among the subfamily Cyprininae.     

Chromosome synapsis and recombination during meiotic division in gynogenetic triploid ginbuna,Carassius auratus langsdorfii

Electron microscope examinations of primary oocytes of gynogenetic triploid ginbuna (Carassius auratus langsdorfii) and diploid gengoroubuna (Carassius auratus cuvieri) were carried out. Typical synaptonemal complexes were observed in both subspecies. In addition, clear differences in PGM-A electrophoretic patterns between a female parent ginbuna and her offspring were detected. It was concluded that synapsis and recombination occur between at least some homologous chromosomes in triploid ginbuna.     

Genetic constitution and phylogenetic relationships of Japanese crucian carps (Carassius)

The genetic constitution and phylogenetic relationships among the proposed species and subspecies of the crucian carp complex in Japan (Carassius cuvieri, C. auratus subspp. 1 and 2, C. a. grandoculis, C. a. buergeri, and C. a. langsdorfii) were investigated based on analyses of the partial nucleotide sequences of the mitochondrial DNA control region and amplified fragment length polymorphisms of nuclear DNA. Our results demonstrate that C. cuvieri and C. auratus are different entities. However, although several distinct lineages were observed for C. auratus, none corresponded to the proposed subspecies. Moreover, the five subspecies of C. auratus were not necessarily separated genetically from each other. Triploid fish, which are currently classified as a single subspecies (C. a. langsdorfii) but are sometimes treated as an independent species, fell into various clades along with diploid individuals of other subspecies that have the same or similar mitochondrial haplotypes. This suggests that gynogenetic triploid crucian carps distributed throughout the Japanese Archipelago have polyphyletic maternal origins. Our results indicate that Japanese crucian carps are a much more complex assemblage than previously believed.     

Genetic variations estimated from PCR-RFLP analysis of two morphs of the freshwater goby <Emphasis Type="Italic">Rhinogobius</Emphasis> in the Lake Biwa water system

The freshwater goby Rhinogobius is the most abundant fish in the shore area of Lake Biwa, Japan. Recently, it has been reported that two morphs of Rhinogobius inhabit this lake. In this study, genetic variations in Rhinogobius sp. OR (Orange-type) and Rhinogobius sp. BW (Biwa-type) in the Lake Biwa water system have been investigated using polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) analysis of mitochondrial DNA, including the variable D-loop region. Samples of Rhinogobius sp. OR were collected from the middle sites of three rivers, two outlets, and two lakeshore sites, whereas samples of Rhinogobius sp. BW were taken from two lakeshore sites. Rhinogobius sp. OR and Rhinogobius sp. BW did not share any haplotypes, suggesting that PCR-RFLP analysis is effective for distinguishing between these species of goby. The haplotype diversity of Rhinogobius sp. OR (0.214–0.543) was lower than that of Rhinogobius sp. BW (0.543–0.682). There were no significant differences in haplotype frequencies between Rhinogobius sp. BW groups from the two localities. In addition, haplotype frequencies in Rhinogobius sp. OR did not differ significantly among samples from the middle sites of rivers, the outlets, and the shores. These results indicate that in Rhinogobius sp. OR there is frequent gene flow among fish inhabiting different sites, and that this species of goby consists of a single population throughout the Lake Biwa water system.     

Difference in the hypoxia tolerance of the round crucian carp and largemouth bass: implications for physiological refugia in the macrophyte zone

The hypoxia tolerance of larval and juvenile round crucian carp, Carassius auratus grandoculis, and largemouth bass, Micropterus salmoides, was determined using respirometry to examine the potential of hypoxic areas in the macrophyte zone as physiological refugia for round crucian carp. The tolerance, which was measured as the critical oxygen concentration (Pc), was 1.32 mg O₂/l in the round crucian carp and 1.93 mg O₂/l in the largemouth bass. As the round crucian carp tolerated hypoxia better than the largemouth bass, hypoxic areas in the macrophyte zone might function as physiological refugia for round crucian carp.     

Molecular characterization of larval anisakid nematodes from marine fishes off the Moroccan and Mauritanian coasts

A total of 242 larval forms of Anisakis collected from marine fishes at different sites off the Moroccan and Mauritanian coasts, recognised as belonging to Type I and Type II larvae, were identified by PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphisms) of the ITS (Internal Transcribed Spacers) region (ITS-1, 5.8 subunit rRNA gene and ITS-2), using a previously established molecular key. The Type I larvae were found with a frequency of 98.34% and were identified as belonging to the following species: A. simplex s.str., A. pegreffii, A. simplex s.str/A. pegreffii heterozygote genotypes, A. typica, A. ziphidarum and Anisakis sp. A. The Type II larvae were found to belong to A. physeteris, with the frequency of 1.65%. The results reported in the present study provide further epizootiological and biological data on the Anisakis spp. in marine fishes off the Moroccan and Mauritanian coasts, improving the picture of the occurrence of these species in the central Atlantic coasts.     

Genetic diversity and divergence of the endangered freshwater goby Rhinogobius sp. BB in Okinawa Island

The genetic diversity and population structure of Rhinogobius sp. BB, the fluvial landlocked endangered goby, were investigated and were compared with those of the closely related amphidromous Rhinogobius sp. MO at 11 microsatellite loci. Specimens of Rhinogobius sp. BB were collected from the Genka and Takae-A rivers, and those of Rhinogobius sp. MO were collected from the Genka and Suginda rivers in Okinawa Island. At 11 microsatellite loci, the two populations of Rhinogobius sp. BB showed lower variation than the two Rhinogobius sp. MO populations: the average number of alleles was 3.6 and 2.0 vs. 8.6 and 7.6, respectively; and the observed heterozygosity was 0.263 and 0.281 vs. 0.440 and 0.545, respectively. Pairwise F_st tests showed significant differences (P < 0.001) among the populations: F_st was 0.525 between the two Rhinogobius sp. BB populations, 0.079 between the two Rhinogobius sp. MO populations, and varied from 0.456 to 0.462 for comparisons among Rhinogobius sp. BB and MO. Nei’s genetic distance between the two Rhinogobius sp. BB populations is extremely large (0.604) compared with that between the two Rhinogobius sp. MO populations (0.126). The two populations of Rhinogobius sp. BB are genetically divergent, and they have extremely low genetic diversity. Therefore, the conservation of Rhinogobius sp. BB in Okinawa Island requires the assessment of each river’s population.     

Fertility of triploid backcross progeny, (GengoroubunaCarassius auratus cuvieri ♀ × carpCyprinus carpio ♂)F11 ♀ × carp or gengoroubuna ♂

The fertility of triploid backcross progeny, (gengoroubunaCarassius auratus cuvieri ♀ × carpCyprinus carpio ♂) F₁ ♀ × gengoroubuna or carp ♂ is reported. In females, spawning could be induced by the injection of pituitary extracts. The fertilization rates were from 52.4 to 88.6%, and free swimming rates from 2.6 to 34.4%. Many normal larvae were obtained after crossing with the sperm of gengoroubuna. However, after crossing with sperm of the loachMisgurnus anguillicaudatus, most of the embryos died before hatching and no normal larvae were obtained. Sperm could not be stripped from the progeny of either type of triploid backcross after the injection of pituitary extracts. Under light microscope, each seminal lobule in the testis was mainly occupied by spermatocytes in various stages. Some spermatids were found in part of the lobules, but no spermatozoa were observed.     

Mitochondrial DNA divergence in yoshinobori gobies (Rhinogobius species complex) between the Bonin Islands and the Japan–Ryukyu Archipelago

To investigate the genetic differentiation between the Bonin (Ogasawara) Islands' freshwater goby Rhinogobius sp. Bonin Island (BI) form (Ogasawara-yoshinobori) and the Japan–Ryukyu Archipelago relatives, the mitochondrial DNA (mtDNA) phylogeny of Japanese Rhinogobius species was inferred from partial nucleotide sequences of the mitochondrial NADH dehydrogenase 5 subunit (ND5) gene (945 bp). The resultant tree showed that the Bonin Islands group separated first from the other Japanese lineage, and a test calculation indicated the divergence date to be approximately 3 million years BP. Although it is necessary to use a more reliable estimate to confirm the divergence date, Rhinogobius sp. BI has retained its mtDNA lineage in the islands for millions of years.     

Physical changes in specimens of five species of Cyprinidae preserved in ethanol and frozen

Preservation in 30% ethanol and freezing to a temperature of ?20 ± 2° C is an appropriate method for measurement of fish eggs, larvae and juveniles. Egg diameter of the common carp Cyprinus carpio increased insignificantly by 1·32% after preservation compared with live size. The total length (L_T) of 1 day post‐hatching (dph) larvae as well as the standard length (LS) of 16 dph larvae of C. carpio increased significantly (2·95 and 1·50%, respectively) after preservation. Egg diameter as well as the L_T of 1 dph larvae of barbel Barbus barbus increased significantly after preservation, by 1·74 and 1·96%, respectively over their original size. The standard length (L_S) of 14 dph larvae of B. barbus as well as juveniles of B. barbus, crucian carp Carassius carassius, common nase Chondrostoma nasus and tench Tinca tinca decreased significantly after preservation (?0·56 to ?5·54%), whereas their body mass increased significantly (11·46–18·57%). Preserved eggs of C. carpio and B. barbus were hard, round and transparent. The larvae and juveniles of examined fishes, preserved in frozen ethanol, were straight, flexible and easily measurable after 60 days. Integrity of body surface and fins, as well as preservation of colours were much better in larvae or juveniles frozen and thawed only once than in specimens frozen and thawed thrice. Cooling in 30% ethanol to a temperature of 6 ± 2° C and freezing in water to a temperature of ?20 ± 2° C are not appropriate preservation methods for eggs and larvae of C. carpio (1 and 16 dph).     

Evidence for the paternal mitochondrial DNA in the crucian carp-like fish lineage with hybrid origin

In terms of taxonomic status, common carp(Cyprinus carpio, Cyprininae) and crucian carp(Carassius auratus, Cyprininae) are different species; however, in this study, a newborn homodiploid crucian carp-like fish(2n=100)(2nNCRC) lineage(F1–F3) was established from the interspecific hybridization of female common carp(2n=100)×male blunt snout bream(Megalobrama amblycephala, Cultrinae, 2n=48). The phenotypes and genotypes of 2 n NCRC differed from those of its parents but were closely related to those of the existing diploid crucian carp. We further sequenced the whole mitochondrial(mt) genomes of the 2n NCRC lineage from F1 to F3. The paternal mt DNA fragments were stably embedded in the mt-genomes of F_1–F_3 generations of 2n NCRC to form chimeric DNA fragments. Along with this chimeric process, numerous base sites of F1–F3 generations of 2 n NCRC underwent mutations. Most of these mutation sites were consistent with the existing diploid crucian carp. Moreover, the mt DNA organization and nucleotide composition of 2n NCRC were more similar to those of the existing diploid crucian carp than those of the parents. The inheritable chimeric DNA fragments and mutant loci in the mt-genomes of different generations of 2nNCRC provided important evidence of the mt DNA change process in the newborn lineage derived from hybridization of different species. Our findings demonstrated for the first time that the paternal mt DNA were transmitted into the mt-genomes of homodiploid lineage, which provided new insights into the existence of paternal mt DNA in the mt DNA inheritance.     

Ecology and impact of Allorhogas sp. (Hymenoptera: Braconidae) and Apion sp. (Coleoptera: Curculionoidea) on fruits of Miconia calvescens DC (Melastomataceae) in Brazil

Two fruit-feeding insects, a gall wasp, Allorhogas sp. (Hymenoptera: Braconidae), and a beetle, Apion sp. (Coleoptera: Curculionoidea), were evaluated in their native habitat in Brazil as potential biological control agents of Miconia calvescens DC (Melastomataceae). Allorhogas sp. occurred at two out of three field sites with native populations of M. calvescens, and Apion sp. occurred at all three sites. Both species exhibited aggregated distributions among M. calvescens trees sampled at each site. Allorhogas sp. infested 9.0% and 3.8% of fruits at each of two sites. The number of larvae and pupae of Allorhogas sp. and/or an unidentified parasitoid (Hymenopetera: Eulophidae: Tetrastichinae) ranged from one to five per infested fruit. Fruits infested with Allorhogas sp. were 20% larger and had 79% fewer seeds than healthy fruits. Although adults of Apion sp. were found on leaves and inflorescences of M. calvescens at all three sites, larvae and pupae were found in fruits at only one site, where a maximum of 1.4% of fruits were infested. Fruits infested by Apion sp. contained only one larva or pupa, and were 15% smaller and had 62% fewer seeds than healthy fruits. While a variety of apionids have been used for biological control in the past, this is the first time a braconid wasp has been considered for biological control of a weed.     

Diet,food preference,and algal availability for fishes and crabs on intertidal reef communities in southern California

Synopsis Herbivory by wide-ranging fishes is common over tropical reefs, but rare in temperate latitudes where the effects of herbivorous fishes are thought to be minimal. Along the west coast of North America, herbivory by fishes on nearshore reefs is largely restricted to a few members of the Kyphosidae, distributed south of Pt. Conception. This paper presents information on natural diets and results from feeding choice experiments for two abundant kyphosids from intertidal habitats in San Diego, California —Girella nigricans andHermosilla azurea, and similar data for the lined shore crab,Pachygrapsus crassipes, which also forages over intertidal reefs. These results are compared with the availability of algae in intertidal habitats measured during summer and winter, on both disturbed and undisturbed habitats. The diets of juveniles ofG. nigricans andH. azurea collected from nearshore habitats were dominated by animal prey (mainly amphipods), but adults of these fishes, andP. crassipes, consumed algae nearly exclusively, with 26, 10, and 14 taxa of algae identified fromG. nigricans, H. azurea, andP. crassipes, respectively. Algae with sheet-like morphologies (e.g.Ulva sp.,Enteromorpha sp., members of the Delesseriaceae) were the principal algae in the diets of the fishes, and calcareous algae (e.g.Corallina sp.,Lithothrix aspergillum) and sheet-like algae (Enteromorpha sp.) comprised the greatest identifiable portion of the shore crab's diet. Feeding choice experiments indicated that the fishes preferred filamentous algae (e.g.Centroceras clavulatum, Polysiphonia sp.,Chondria californica) and sheet-like algae (e.g.Enteromorpha sp.,Ulva sp.,Cryptopleura crispa) over other algal morphologies, whereas the shore crab chose jointed calcareous algae (e.g.Lithothrix aspergillum, Corallina vancouveriensis, Jania sp.) most frequently. The diets and preferences for algae by the fishes were generally most similar to the assemblage of algae available in early successional (disturbed) habitats during summer when sheet-like and filamentous algae are abundant. The shore crab exhibited the opposite trend with a diet more similar to late successional (undisturbed) habitats.     

Parasitoids and other natural enemies of Maruca vitrata feeding on Sesbania cannabina in Taiwan

Natural enemies of Marucavitrata (Geyer) feeding concealed inside leafwhorls of a green manure legume, Sesbaniacannabina, were monitored during the 1996 and1997 summer seasons in Taiwan. A braconid,Apanteles taragamae, accounted for anaverage of 92% of all parasitoid specimensreared from M. vitrata larvae and pupae.Other parasitoid species included two otherbraconids: Bassus asper and Dolichogenidea sp., three ichneumonids: Trichomma sp., Triclistus sp., and Plectochorus sp., and two unidentifiedtachinids. All parasitoid species attacked M. vitrata larvae and emerged as mature larvaefrom host larvae (braconids) or as adults fromhost pupae (ichneumonids and tachinids). Oneunidentified predatory staphylinid and threeentomopathogenic fungi: Fusarium sp.Paecilomyces sp. and Beauveriabassiana were also observed attacking M.vitrata larvae. Apanteles taragamae is a solitary endoparasitoid. Its parasitismreached as high as 63% of M. vitratalarvae found attacking S. cannabina. Theparasitism was higher during June to August andreduced during September to November when otherparasitoid species were more active. Ninespecies of hyperparasitoids, all belonging toHymenoptera, were recovered, mainly from thebraconids.     

Toward a preliminary assessment of the diversity and origin of Cyprinid fish genus Carassius in Iran

Introduction of non-native fish species has been criticized as one of the major threats to freshwater ecosystems. During recent years, the cyprinid fish genus Carassius has been the subject of various studies in terms of introduction biology and ecology. The taxonomy and identification of described species in the genus Carassius is still ambiguous. In order to increase the knowledge of introduced species of the genus in Iran, molecular approaches were employed to identify species based on the mitochondrial gene, cytochrome b (Cyt b). In total, 417 specimens across Eurasia and some other parts of the world have been analyzed. Afterwards, investigation of the possible origin and pathways of introduction was done using analysis of haplotype networks based on the Cyt b gene. The results revealed that there are three introduced species i.e., Carassius auratus, Carassius gibelio and Carassius langsdorfii, in inland waters of Iran. Carassius langsdorfii was recorded from Iran and western Asia for the first time. Analysis of haplotype network showed various potential sources of introduction for each species in Iran and regions beyond their native distribution ranges.     

Coloration and ontogenetic features of fluviatile species of <Emphasis Type="Italic">Rhinogobius</Emphasis> (Gobioidei: Gobiidae) in Amami-oshima Island,Ryukyu Islands,Japan

 The fluviatile Rhinogobius sp. 1 (hereinafter R. sp. 1) (Japanese name kibara-yoshinobori or English name yellow belly medium-egg type) is broadly distributed in the Ryukyu Islands ranging from Amami-oshima Island to Iriomote Island. However, based on the molecular evidence, this species has probably had multiple origins from amphidromous species. We compared the coloration of specimens at Amami-oshima I. with that of the other specimens (islands) in the Ryukyu Is. Amami-oshima I. specimens of R. sp. 1 were different from the specimens of other populations in having five to six cross lines on the caudal fin. We examined the early life history of fluviatile Rhinogobius at Amami-oshima I., based on rearing experiments, and estimated the age of wild specimens by counting otolith increments. Spawned egg capsules of “Rhinogobius sp. 1” at Amami-oshima I. were larger than those of amphidromous species. Their developmental stage in newly hatched larvae was similar to that of Okinawan R. sp. 1 in having a straight notochord; however, it had a flexed notochord earlier than that of Rhinogobius sp. 2 (hereinafter R. sp. 2) (aobara-yoshinobori or blue belly medium-egg type). The ontogenetic trajectory of “Rhinogobius sp. 1” in Amami-oshima I. was similar to Okinawan R. sp. 1 (30 days in larval duration) but different from R. sp. 2 (15 days). In amphidromous relatives of R. sp. 2, developmental timing during larval period was earlier than in the amphidromous relatives of R. sp. 1. It has been suggested that the phenomenon of fluvial landlocking has been accompanied somewhat by egg size increase (on the basis of a few examples). However, the ontogenetic trajectories of fluviatile Rhinogobius species may be a reflection of those of ancestral amphidromous species. Received: March 18, 2002 / Revised: November 7, 2002 / Accepted: December 10, 2002     

Stable Isotope Analysis of Amphidromous Hawaiian Gobies Suggests Their Larvae Spend a Substantial Period of Time in Freshwater River Plumes

Synopsis We employed stable isotope analysis (δ¹³C, δ¹⁵N) to evaluate the sources of nutrients used by amphidromous gobiid fishes (Lentipes concolor, Sicyopterus stimpsoni, Awaous guamensis) caught migrating into and living in Hakalau Stream, Hawaii. Although considerable variation amongst the stable isotope values of stream items was noted across all 4 years of our study, the relationships between the fishes were relatively constant. Stable isotope values of recruiting gobies were consistently closer to those of both inshore plankton and freshwater adults than those of offshore plankton, suggesting that the larvae of these species derive much of their nutrition from inshore environments influenced by fresh water. Small differences between the stable values of these species further suggested that their larvae come from different inshore locations. After entering fresh water all species appear to swim rapidly upstream without feeding. Finally, once well upstream, adult L. concolor and A. guamensis appear to assume an omnivorous diet while adult S. stimpsoni rely upon autochthonous production within streams. We propose that freshwater food webs play an integral yet complex role in the lives of both larval and adult amphidromous Hawaiian fishes.     

Genetic differentiation among Greek lake populations of Carassius gibelio and Cyprinus carpio carpio

The genetic structure of the Western Greece lake populations of Carassius gibelio and Cyprinus carpio carpio populations was characterized by using a PCR-based RFLP and sequencing analysis of mitochondrial rDNA genes and regions (16S rDNA, cytochrome b and D-loop). Our analysis was able to detect: (a) two haplotypes in C. c. carpio populations and two haplotypes in C. gibelio populations (b) a high nucleotide divergence between the two species and (c) two genetically distinct C. gibelio populations, one existing in the Amvrakia habitat (AMV1) with a second in Ozeros and Trichonida (OZE1 and TRI1) habitat. The present analysis indicates that genetic diversity observed was limited with a haplotype index between 0.0 and 55.6%, and a nucleotide diversity within and among populations between 0.0 and 1.27%. It also underlines a restricted mtDNA-based evaluation of the phylogenetic relationships among C. gibelio and C. c. carpio populations. In addition, the present study contributed knowledge on the genetic variation and structure of these populations which is absolutely necessary for any efficient fish management and/or conservation programme.     

Early morphological development ofOmobranchus fasciolatoceps andO. Punctatus (Blenniidae: Omobranchini) reared in an aquarium

Larval and juvenile development of two blenniids,Omobranchus fasciolatocepts andO. punctatus, is described using eggs collected from natural waters in Tokyo Bay and incubated in an aquarium. These larvae and juveniles are compared with those of two otherOmobranchus species,O. elegans andO. loxozonus, distributed widely in Japan.Onobranchus punctatus is characterized by a unique, pointed snout in preflexion larvae, no melanophores proximally on the lower part of the pectoral fins in flexion and postflexion larvae, and pterygiophores projecting externally as blades between the dorsal and anal fin-rays in postflexion larvae and juveniles.Omobranchus fasciolatoceps has the following characteristics: a few melanophores on the fore-and mid-brain, but none on the hind-brain in preflexion larvae; no melanophores on the cleithral symphysis in flexion and postflexion larvae; no external pterygiophore blades in postflexion larvae and juveniles; and a unique dorsal skin flap on the head in juveniles. Ontogenetic developement of dorsal and anal pterygiophores is described forO. fasciolatoceps andO. punctatus. InO. punctatus, the postero-distal part of each proximal radial projects remarkably to form the external blades between the soft fin-rays, whereas the external blades between the fin spines are formed by fusion of a dermal bone developed from the antero-distal part of each proximal radial with the adjacent distal radial.     

Fish entrapment in the seawater intake of a power plant at Karachi coast

Synopsis The study deals with the entrapment of fishes in the intake of Karachi Nuclear Power Plant. A total of 62 species of marine fishes belonging to 43 genera were observed entrapped in the seawater intake. Therapon puta, Liza waigiensis, Abudefduf septemfasciatus, and Lagoceptalus inermis were the most common. Mass mortalities of the juveniles of Sardinella sindensis were recorded in September and October of 1974, 1975, and 1977. Commercially important fishes such as sardines, mullets and anchovies, were entrapped in the intake of the power plant in considerable numbers at various times of the year. The majority of fishes removed from the screen washes of the power plants were killed by impingement.