Genetic variation of the cytochrome b gene in the rosy bitterling, Rhodeus ocellatus (Cyprinidae) in Japan

According to conventional views, the rosy bitterling, Rhodeus ocellatus, comprises two subspecies, R. ocellatus kurumeus and R. ocellatus ocellatus, the former being native to Japan whereas the latter was introduced into Japan from China during World War II. To examine the genetic structure of Japanese R. ocellatus, part of the mitochondrial cytochrome b gene from 48 individuals collected from various locations in Japan was sequenced. Three major mitochondrial lineages were found. Based on historical evidence, two of these represent R. ocellatus ocellatus and the third R. ocellatus kurumeus. The existence of two distinct lineages of R. ocellatus ocellatus in Japan suggests at least two colonizations. Some local populations comprised purely R. ocellatus kurumeus, but those from Kashima and Ogori included both subspecies. Because the proportion of R. ocellatus ocellatus in Kashima increased from 1994 to 1995, invasion by R. ocellatus ocellatus into R. ocellatus kurumeus habitats is apparently in progress. Received: April 30, 1999 / Revised: March 22, 2000 / Accepted: December 20, 2000     

Algivore or Phototroph? Plakobranchus ocellatus (Gastropoda) Continuously Acquires Kleptoplasts and Nutrition from Multiple Algal Species in Nature

The sea slug Plakobranchus ocellatus (Sacoglossa, Gastropoda) retains photosynthetically active chloroplasts from ingested algae (functional kleptoplasts) in the epithelial cells of its digestive gland for up to 10 months. While its feeding behavior has not been observed in natural habitats, two hypotheses have been proposed: 1) adult P. ocellatus uses kleptoplasts to obtain photosynthates and nutritionally behaves as a photoautotroph without replenishing the kleptoplasts; or 2) it behaves as a mixotroph (photoautotroph and herbivorous consumer) and replenishes kleptoplasts continually or periodically. To address the question of which hypothesis is more likely, we examined the source algae for kleptoplasts and temporal changes in kleptoplast composition and nutritional contribution. By characterizing the temporal diversity of P. ocellatus kleptoplasts using rbcL sequences, we found that P. ocellatus harvests kleptoplasts from at least 8 different siphonous green algal species, that kleptoplasts from more than one species are present in each individual sea slug, and that the kleptoplast composition differs temporally. These results suggest that wild P. ocellatus often feed on multiple species of siphonous algae from which they continually obtain fresh chloroplasts. By estimating the trophic position of wild and starved P. ocellatus using the stable nitrogen isotopic composition of amino acids, we showed that despite the abundance of kleptoplasts, their photosynthates do not contribute greatly to the nutrition of wild P. ocellatus, but that kleptoplast photosynthates form a significant source of nutrition for starved sea slugs. The herbivorous nature of wild P. ocellatus is consistent with insights from molecular analyses indicating that kleptoplasts are frequently replenished from ingested algae, leading to the conclusion that natural populations of P. ocellatus do not rely on photosynthesis but mainly on the digestion of ingested algae.     

Evaluation of Morphological Variation in the Lichen Diploschistes ocellatus (Ascomycota, Ostropales): Evidence from Nuclear rDNA ITS Sequence Data

Abstract: A morphologically distinct population (var. almeriensis ) of the lichen species Diploschistes ocellatus from southern Spain is examined using morphological and molecular methods. While the morphological examination revealed some unique features of var. almeriensis, the ITS sequence data did not support a distinction from D. ocellatus s. str. The Kishino-Hasegawa test did not rule out trees distinguishing between the two morphotypes, although spectral analytical data support an inclusion of var. almeriensis in D. ocellatus. The influence of potentially erronously aligned ambiguous sites is evaluated by analysis of different alignments. However, all alignments agree in placing all specimens of D. ocellatus together and, consequently, in the consensus tree D. ocellatus s. lat. forms a monophyletic clade with high bootstrap support.     

Gastrointestinal helminths of the lizard Chalcides ocellatus from Benghazi, Libya

Chalcides ocellatus, a scincid lizard, sampled during October 1998 to December 1999 from Benina (farmland) and Al-Kueffia (a rocky area) in Benghazi, Libya, was found to harbour three intestinal nematodes, Pharyngodon mamillatus, Thelandros alatus and Parapharyngodon micipsae, and an intestinal cestode, Oochoristica tuberculata. Thelandros alatus, P. micipsae and O. tuberculata were recorded for the first time in C. ocellatus. Parapharyngodon micipsae was detected in C. ocellatus from Benina and O. tuberculata from lizards in Al-Kueffia. The majority (87.6%) of C. ocellatus were infected with helminth parasites and the levels of infection were higher in males than in females but this difference was not significant. Pharyngodon mamillatus recorded the highest number of nematode parasites in C. ocellatus, although there was no relationship between the number of nematode parasites recovered and host density. In lizards infected with the cestode O. tuberculata, males had shorter snout-vent lengths than females. From a total of 153 C. ocellatus, 120 (78.4%) showed single and 14 (9.2%) showed mixed parasitic infections. Mixed infections between different species of nematodes were not observed.     

A morphological and cytochemical study of erythrocytes, thrombocytes and leukocytes in four freshwater teleosts

Erythrocytes, thrombocytes and leukocytes morphology and cytochemical staining were studied in big head carp Aristichthys nobilis , oscar Astronotus ocellatus , traíra Hoplias malabaricus and lambari Astyanax bimaculatus . Reticulocytes contained a granular material similar to residual RNA following staining with brilliant cresyl blue. Neutrophils, lymphocytes and monocytes were morphologically similar in all the four species. Thrombocytes were present in all the four species and were predominantly fusiform, whereas eosinophils occurred only in A. ocellatus . Aristichthys nobilis contained a leukocyte with unstained granules following Romanowsky-type staining, which stained intensely with periodic acid-Schiff (PAS). Glycogen granules were present in thrombocytes, neutrophils and eosinophils but not in monocytes or lymphocytes. Peroxidase staining was observed in neutrophils of A. ocellatus , H. malabaricus and A. bimaculatus but not in A. nobilis . Monocytes of A. ocellatus , H. malabaricus and A. bimaculatus stained positively for non-specific esterase, whereas those of A. nobilis did not stain. Thrombocytes and leukocytes in all four species were negative for alkaline phosphatase. Neutrophils of A. ocellatus and H. malabaricus may be involved in respiratory burst and may play an important microbicidal role.     

Genetic markers distinguishing between the two subspecies of the rosy bitterling,Rhodeus ocellatus (Cyprinidae)

Eleven populations of the rosy bitterling,Rhodeus ocellatus, from different localities in Japan, were genetically compared at 16 protein-coding loci using starch-gel electrophoresis. Two loci,Ldh-2 andPgdh, were demonstrated as diagnostic markers for the identification of two subspecies;R. ocellatus kurumeus, which is native to Japan, andR. ocellatus ocellatus, which was introduced from China. The two subspecies were distinguished by the complete substitution of different alleles between them. Population ofR. ocellatus kurumeus occurring in Yao City, Osaka, and in Kanzaki, Saga Prefecture were genetically closely related to each other (genetic distance: D=0.056) but distantly so toR. ocellatus ocellatus from Saitama Prefecture (D=0.202 or 0.265). Electrophoretic analyses also elucidated the existence of hybrid populations of the two subspecies. The populations ofR. ocellatus kurumeus in Yao City had lower genetic variability and a lower incidence of white coloration on the ventral fins than populations of the same in Saga Prefecture. The present study strongly implies that the introduction of the foreign freshwater fishes with subspecific differentiation, into the original range of indigenous subspecies, should be averted not to bring the genetic pollution.     

Isolation and characterization of new microsatellite markers for rose bitterlings, Rhodeus ocellatus

The Japanese rose bitterling (Rhodeus ocellatus kurumeus) is facing imminent extinction because of hybridization and competition from an invasive alien subspecies (Rhodeus ocellatus ocellatus). Eleven new microsatellite markers for the two subspecies were developed using dinucleotide repeat specific polymerase chain reaction. The number of alleles per locus and the heterozygosity in R. o. kurumeus were lower than those in R. o. ocellatus. Most of these microsatellite markers were successfully cross‐amplified in three Acheilognathinae species.     

Spatial Homogeneity and Temporal Heterogeneity of Red Drum (Sciaenops ocellatus) Microsatellites: Effective Population Sizes and Management Implications

The red drum (Sciaenops ocellatus) is one of a number of species that occupy estuarine waters as juveniles and migrate to open ocean waters as adults. This species has experienced dramatic declines in population numbers over the past 2 decades, which has prompted increasing fishery restriction. In addition, hatchery augmentation has been initiated by several states to increase the abundance of juveniles in local areas. In South Carolina hatchery-reared fish have made significant (20%) contributions to the juvenile population on very local scales. As hatchery-reared fish are typically produced by a small number of individuals, the genetic consequences of augmentation programs are of concern. In this article we assess genetic variation at 5 microsatellite loci in S. ocellatus. The data indicate little geographic differentiation among samples collected along the Atlantic Coast of the United States, but substantial differences among year classes taken from South Carolina. The gene frequency differences among year classes were used to estimate the effective population size (Nc) of S. ocellatus in South Carolina and suggested that Ne was less than 300 from 1990 to 1993 and increased to about 1000 in 1994 and 1995. Whether this increase reflects the effectiveness of management regulations or simply a random fluctuation in S. ocellatus populations is not clear. The data suggest that a limited number of individuals produce the bulk of a given year class and support the sweepstakes hypothesis. Given the small Ne and estimates of the contribution of hatchery-reared fish to the wild stock, it is suggested that programs have the potential to increase, rather than decrease; Ne in the wild.     

我国缅阴虫属的分布及眼斑缅阴虫的生物学特性

缅阴虫属(Burmoniscus)是我国长江以南热带和亚热带地区中重要的陆栖等足类。本文对我国该属的种类组成和分布做了总结和分析,并对该属的代表种类——眼斑缅阴虫(B.ocellatus)的栖息地、食性、食量及繁殖季节等生物学特性进行了研究和综述。     

Low genetic variation and inbreeding depression in small isolated populations of the Japanese rosy bitterling, Rhodeus ocellatus kurumeus

The Japanese rosy bitterling, Rhodeus ocellatus kurumeus, has been affected not only by the invasion of another subspecies, R. o. ocellatus, from China, but also by habitat fragmentation. In this study, the effects of habitat fragmentation on the fitness of R. o. kurumeus were investigated. Owing to exclusion by R. o. ocellatus, R. o. kurumeus in Honshu and Shikoku has disappeared entirely, except for small populations in isolated man-made ponds in Osaka and Kagawa. In Kyushu it still occupies open water systems, into which R. o. ocellatus has only recently invaded. Meristic and genetic data show that the diversity of R. o. kurumeus is significantly lower in the isolated Osaka and Kagawa populations than the non-isolated Fukuoka population. The Osaka population is inferior to the Fukuoka population in terms of viability and growth. The viability of reciprocal inter-population hybrids between the Osaka and Fukuoka populations was, however, as high as that of the Fukuoka population. In addition to the high scores of band sharing index (BSI) in RAPD-PCR analysis, acceptance of transplanted scales among individuals, irrespective of natal pond, indicates that the Osaka population forms a highly inbred line. These results suggest that low genetic variation is associated with inbreeding depression in the small isolated Osaka populations. Consequently, the management of ponds, including the free movement of individuals, in addition to measures to prevent the invasion of R. o. ocellatus, is necessary for the conservation of R. o. kurumeus.     

Cytogenetic and random ampliied polymorphic DNA analysis of Leptodactylus species from rural and urban environments (Anura, Amphibia)

Cytogenetic and random amplified polymorphic DNA analyses carried out in the species Leptodactylus podicipinus, L. ocellatus, L. labyrinthicus, and L. fuscus from rural and urban habitats of the northwest region of S?o Paulo State, Brazil, showed that the karyotypes (2n = 22), constitutive heterochromatin distribution and nucleolus organizer region (NOR) location did not differ between the populations from the two environments. The in situ hybridization with an rDNA probe confirmed the location of the NORs on chromosome 8 revealing an in tandem duplication of that region in one of the chromosomes of L. fuscus. DAPI showed that part of the C-band-positive heterochromatin is rich in AT, including that in the proximity the NORs in L. podicipinus and L. ocellatus. The molecular analyses showed that the two populations (urban and rural) of L. podicipinus and L. fuscus are similar from a genetic point of view. The urban and rural populations of species L. ocellatus and L. labyrinthicus showed differences in genetic structures, probably due to urbanization which interferes with the dispersion of those frogs. The marked differences observed between the two populations of L. ocellatus can be representing the cryptic condition of the species. Unweighted pair-group method of analysis and genetic distance analysis detected the genetic proximity between L. ocellatus and L. fuscus. The results indicate that there was no reduction in the genetic diversity in the populations from the urban environment; however, the survival of these frogs would not be guaranteed in the case of an increase in human impact especially for populations of L. labyrinthicus and L. ocellatus.     

Placental ontogeny of the Tasmanian scincid lizard, Niveoscincus ocellatus (Reptilia: Squamata)

A prominent scenario for the evolution of reptilian placentation infers that placentotrophy arose by gradual modification of a simple vascular chorioallantoic placenta to a complex structure with a specialized region for nutrient transfer. The structure of the chorioallantoic placenta of Niveoscincus ocellatus, apparently described originally from a single embryonic stage, was interpreted as a transitional evolutionary type that provided support for the model. Recently, N. ocellatus has been found to be as placentotrophic as species with complex chorioallantoic placentae containing a specialized region called a placentome. We studied placental development in N. ocellatus and confirmed that the chorioallantoic placenta lacks specializations found in species with a placentome. We also found that this species has a specialized omphaloplacenta. The chorioallantoic placenta is confined to the region adjacent to the embryo by a membrane, similar to that found in some other viviparous skinks, that divides the egg into embryonic and abembryonic hemispheres. We term this structure the "inter-omphalopleuric" membrane. The position of this membrane in N. ocellatus is closer to the embryonic pole of the egg than to the abembryonic pole and thus the surface area of the omphaloplacenta is greater than that of the chorioallantoic placenta. In addition, the omphaloplacenta is regionally diversified and more complex histologically than the chorioallantoic placenta. An impressive and unusual feature of the omphaloplacenta of N. ocellatus is the development of extensive overlapping folds in the embryonic component of mid-gestation embryos. The histological complexity and extensive folding of the omphaloplacenta make this a likely site of placental transfer of nutrients in this species.     

久效磷对美国红鱼鳃Na^＋／K^＋-ATP酶活性和超显微结构的影响

经不同试验浓度的久效磷(0．25、0．5、1．0和2．0mg·L^-1)处理美国红鱼4d后,分别对鱼鳃Na^+／K^+—ATP酶活性和氯细胞密度进行了测定和计数,并观察了鱼鳃组织显微结构和超微结构的变化。结果表明,低浓度久效磷(0．25mg·L^-1)处理可以诱导鱼鳃氯细胞大量增生,Na^+／K^+—ATP酶活性增强,随着试验浓度的增加,久效磷对鳃组织的损伤越来越重,Na^+／K^+—ATP酶活性逐渐降低;久效磷对鱼鳃显微结构的损伤表现为鳃小片上皮细胞水肿、脱离。鳃小片基部粘连。鳃小片上皮细胞角质化;超微结构变化主要为内质网、线粒体、微小管和核膜的水肿及部分溶解,这种损伤表现为由细胞膜到细胞核的动态过程。     

Placental nutrition in the Tasmanian skink, Niveoscincus ocellatus

Niveoscincus ocellatus is an important species in historical analyses of the evolution of viviparity because it is the species upon which the type II chorioallantoic placenta was based. Here we describe the net nutrient uptake across the placenta of N. ocellatus for comparison with other species of skinks with complex placentae. N. ocellatus is highly placentotrophic, with neonates being 1.68-times larger in dry matter than the fresh eggs. There is an increase of nitrogen from 6.3 +/- 0.2 mg to 9.2 +/- 0.6 mg, and ash from 3.8 +/- 0.3 mg to 6.7 +/- 0.6 mg. The increase in ash is made up by a more than two-fold increase in the amounts of calcium, potassium and sodium. There is no significant difference in lipids in the neonates compared to fresh eggs, so considerable lipid must have crossed the placenta to provide energy for embryonic development. N. ocellatus is significantly more placentotrophic than Niveoscincus metallicus, which also has a complex chorioallantoic placenta. Discovery of substantial placentotrophy in this genus confirms that two lineages of Australian lygosomine skinks (represented by the genera Pseudemoia and Niveoscincus) have evolved this pattern of embryonic nutrition and supports the hypothesis that the evolution of reptilian placentotrophy involves specialisations in addition to structural modifications of the chorioallantoic placenta.     

Chromosomal studies on five species of the genus Leptodactylus Fitzinger, 1826 (Amphibia, Anura) using differential staining

Cytogenetic studies were carried out on five species of Leptodactylus, namely L. fuscus, L. notoaktites, L. labyrinthicus, L. ocellatus, and L. podicipinus, after standard staining, Ag-NOR and C-banding as well as BrdU incorporation for three of them. The species had 2n = 22 chromosomes and two basic karyotype patterns. Chromosome 8 was a marker bearing a secondary constriction. In all species, this secondary constriction corresponded to the Ag-NOR site. The species had centromeric C-bands in all chromosomes of the complement, but some interstitial or telomeric bands seemed to differentiate some karyotypes, either at the species or the population level. In L. ocellatus, the C-banding pattern confirmed the occurrence of a heteromorphic pericentric inversion in chromosome 8 in specimens from one of the populations. The BrdU incorporation technique showed no detectable difference in the replication patterns of the major bands in the chromosomes of L. notoaktites, L. labyrinthicus, and L. ocellatus.     

Comparative immunochemical study of the serum proteins of Lacertilia (Reptilia)]

The immunochemical relationships of the serum proteins of Uromastix acanthinurus, Agama mutabilis, Oplurus cuvieri, O. quadrimaculatus, Chamaeleo chamaeleon, Chalcides ocellatus and Lacerta lepida were studied by means of immunoelectrophoresis with an antiserum anti-Uromastix successively absorbed. Homogeneity of the Iguania species was pointed out and they were found more closely related to the Scincid Chalcides ocellatus than to the Lacertid Lacerta lepida. Within the Iguania, the two Agamids are immunologically more closely related to the Chamaeleon than to the two Iguanids.     

Cell mediated immune response of Bufo arenarum and Leptodactylus ocellatus (Amphibia, Anura) to mycobacterial antigens

The lymphocyte transformation test was applied to the study of cell mediated immune (CMI) response in vitro to mycobacterial antigens in Bufo arenarum and Leptodactylus ocellatus previously inoculated with a suspension of Mycobacterium tuberculosis. The stimulated samples of blood both in B. arenarum and L. ocellatus showed a significant increase in the percent of blastic dedifferentiation (almost 100% with respect to controls) suggesting the presence of a positive CMI response to PPD. The differences were observed on the fifth day of incubation and lasted until the seventh day.     

Astronotus ocellatus (Cichlidae:Pisces) and Macropodus opercularis (Anabatidae:Pisces) as predators of immature Aedes fluviatilis (Diptera:Culicidae) and Biomphalaria glabrata (Mollusca:Planorbidae).

Two fish species, Astronotus ocellatus (Cichlidae) and Macropodus opercularis (Anabatidae) were tested for predacious behavior toward immature mosquitoes (Aedes fluviatilis, Diptera:Culicidae) and schistosomiasis snail hosts (Biomphalaria glabrata, Mollusca:Planorbidae), in the presence or absence of non-living food and in laboratory conditions. A. ocellatus, a species indigenous to Brazil, was a very efficient predator of both organisms (alpha = 0.05); M. opercularis, an exotic species, preyed well on immature mosquitoes, but small snails and snail egg-masses were ingested only irregularly. Both fish species seemed to prefer live to non-living food.     

中国杉藻科(Gigartinaceae)两个新记录种的研究

报道中国产杉藻科(Gigartinaceae)角叉菜(Chondrus ocellatus)和日本马泽藻(Mazzaella japonica)两个种的分类研究,均为中国新记录。     

Restriction site heteroplasmy in the mitochondrial DNA of the marine fish Sciaenops ocellatus (L.)

Restriction site heteroplasmy involving the enzymes NcoI and XbaI was detected in the mitochondrial DNAs of two individuals of the marine fish Sciaenops ocellatus. This represents only the sixth documented example of mitochondrial DNA restriction site heteroplasmy in animals. Two heteroplasmic individuals were found in a survey of nearly 750 individuals, suggesting that in most studies the incidence of mitochondrial DNA site heteroplasmy may be too low to be routinely detected.