Genetic structure of Chinese sucker population Myxocyprinus asiaticus in the Yangtze River based on mitochondrial DNA marker

The sequencing analysis of the mitochondrial DNA control region (mtCR DNA) was performed to assess the genetic divergence and population structure of the Chinese sucker Myxocyprinus asiaticus (Cypriniformes Catostomidae) using four sample lots from natural populations of the Yangtze River. The mtCR DNA sequences of approximately 920 base pairs were obtained. A total of 223 nucleotide positions were polymorphic, and these defined 39 haplotypes. Of the 39 haplotypes, 37 (90%) were not shared, and among the populations as a whole there was little sharing of haplotypes. The average haplotype diversity (0.958) and the average nucleotide diversity (0.052) indicated a higher level of genetic diversity of Chinese sucker through the river. Analysis of molecular variation (AMOVA) of data revealed significant partitioning of variance (P<0.001) among populations (60.29%), and within populations (39.71%). The topology according to the neighbor joining and maximum parsimony methods showed mosaic composition of the 39 haplotypes, suggesting that the populations wore not completely divergent. The pairwise F statistic values, however, indicated that the population structuring existed to some extent among the geographic populations. There was a positive relationship between the aquatic distance and the genetic distance (Fst) among the populations (P<0.05). Based on our data, it is suggested that genetic drift, gene flow, and stochastic events are the possible factors influencing the population structure and genetic variation.

Differential expression of two Carassius auratus Mx genes in cultured CAB cells induced by grass carp hemorrhage virus and interferon

UV-inactivated GCHV (grass carp hemorrhage virus) is able to induce an antiviral state in cultured CAB cells (crucian carp Carassius auratus blastulae embryonic cells) via the production of interferon (IFN). In the current work, the full-length cDNAs of two Mx genes, termed CaMx1 and CaMx2, have been cloned and sequenced from UV-inactivated GCHV-infected and still IFN-producing CAB cells by suppression subtractive hybridization. Their putative proteins show the characteristically structural features of mammalian IFN-induced Mx proteins, including GTP-binding motif, dynamin family signature and leucine zipper motif. CaMx1 exhibits 85% sequence identity to zebrafish MxA and 72-74% to three Atlantic salmon Mx proteins. CaMx2 is most similar to zebrafish MxE, with 80% identity, and then rainbow trout Mx3, with 52%. Constitutive expression was detected by RT-PCR for CaMx1, but not for CaMx2, in normal CAB cells, but their up-regulations could be induced after treatment with active GCHV, UV-inactivated GCHV and CAB IFN. Distinct kinetics of expression was observed for either CaMx1 or CaMx2 corresponding to the three stimuli, and even between CaMx1 and CaMx2, corresponding to the same stimulus. Upon virus infection, the transcriptional induction was strongly blocked for CaMx2 by cycloheximide (CHX), whereas almost nothing was observed for CaMx1. By contrast, following treatment with CAB IFN, CHX did not inhibit either gene transcription. Collectively, these results suggest that there are very distinct mechanisms for modulating the expression of both CaMx1 and CaMx2 in normal and GCHV-infected CAB cells.

Infection and propagation of lymphocystis virus isolated from the cultured flounder Paralichthys olivaceus in grass carp cell lines

The causative agent of lymphocystis disease that frequently occurs in cultured flounder Paralichthys olivaceus in China is lymphocystis virus (LV). In this study, 13 fish cell lines were tested for their susceptibility to LV. Of these, 2 cell lines derived from the freshwater grass carp Ctenopharyngodon idellus proved susceptible to the LV, and 1 cell line, GCO (grass carp ovary), was therefore used to replicate and propagate the virus. An obvious cytopathic effect (CPE) was first observed in cell monolayers at 1 d post-inoculation, and at 3 d this had extended to about 75% of the cell monolayer. However, no further CPE extension was observed after 4 d. Cytopathic characteristics induced by the LV were detected by Giemsa staining and fluorescence microscopic observation with Hoechst 33258 staining. The propagated virus particles were also observed by electron microscopy. Ultrastructure analysis revealed several distinct cellular changes, such as chromatin compaction and margination, vesicle formation, cell-surface convolution, nuclear fragmentation and the occurrence of characteristic 'blebs' and cell fusion. This study provides a detailed report of LV infection and propagation in a freshwater fish cell line, and presents direct electron microscopy evidence for propagation of the virus in infected cells. A possible process by which the CPEs are controlled is suggested.

Comparative ontogenetic behavior and migration of kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii, from the Amur River

We conducted laboratory experiments with kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii, to develop a conceptual model of early behavior. We daily observed embryos (first life phase after hatching) and larvae (period initiating exogenous feeding) to day-30 (late larvae) for preference of bright habitat and cover, swimming distance above the bottom, up- and downstream movement, and diel activity. Day-0 embryos of both species strongly preferred bright, open habitat and initiated a strong, downstream migration that lasted 4 days (3 day peak) for kaluga and 3 days (2 day peak) for Amur sturgeon. Kaluga migrants swam far above the bottom (150 cm) on only 1 day and moved day and night; Amur sturgeon migrants swam far above the bottom (median 130 cm) during 3 days and were more nocturnal than kaluga. Post-migrant embryos of both species moved day and night, but Amur sturgeon used dark, cover habitat and swam closer to the bottom than kaluga. The larva period of both species began on day 7 (cumulative temperature degree-days, 192.0 for kaluga and 171.5 for Amur sturgeon). Larvae of both species preferred open habitat. Kaluga larvae strongly preferred bright habitat, initially swam far above the bottom (median 50-105 cm), and migrated downstream at night during days 10-16 (7-day migration). Amur sturgeon larvae strongly avoided illumination, had a mixed response to white substrate, swam 20-30 cm above the bottom during most days, and during days 12-34 (most of the larva period) moved downstream mostly at night (23-day migration). The embryo-larva migration style of the two species likely shows convergence of non-related species for a common style in response to environmental selection in the Amur River. The embryo-larva migration style of Amur sturgeon is unique among Acipenser yet studied.

Distribution of epilithic algae in the Xiangxi River system and their relationships with environmental factors

The spatial pattern of epilithic algae in the Xiangxi River system was studied in relation to several environmental factors by two-way indictor species analysis (TWINSPAN), detrended correspondence analysis (DCA), and canonical correspondence analysis (CCA). Eighty-nine taxa including diatoms, green algae, and blue-green algae were observed. Diatoms were dominant, and Cocconeis placentula, Cymbella minuta, Diatoma vulgare, and Gomphonema angustatum appeared in most of sampling sites. By TWINSPAN and DCA, thirty-one sites were divided into three groups based on composition and relative richness of benthic algae. CCA indicated that SiO2, pH, total phosphorus, Ca2+, velocity, elevation, and Cl- were significant environmental factors affecting the distribution of algae communities. In this minimal subset, SiO2 and pH were the most influential variables.

Observations on behavior and ecology of the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis) group at Tian-e-Zhou Oxbow of the Yangtze River

The authors made 39 surveys (a total of 161 days) in the Tian-e-Zhou Oxbow of the Yangtze River, China, for observing 13 Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis) captured from the main stream of the Yangtze River and 7 juveniles born in the oxbow from January 1997 to July 2000. The animals were usually divided into several "core" groups and moved around in shallow, muddy-bottom areas with the largest individual in the lead. Each core group was composed of 2-3 animals (either 2 adults, 1 adult and 1 juvenile, 2 adults and 1 juvenile, or 2 adults and 1 calf). Newly-released animals joined the other animals first, and then reorganized their own groups one or two days later. Average breath interval was 34.4 s (+/- s.d. 4.39) for individuals in the group. The animals mated from May through June and gave birth during the second and last ten days of April of the next year. The gestation period was estimated as 310 - 320 days. Calves over 5 months old began to eat small fish. The distance of calves swimming apart from their suspected mothers increased each month. These findings will help in the management of the reserve to protect this unique freshwater porpoise.

Common evolutionary origin of aquareoviruses and orthoreoviruses revealed by genome characterization of Golden shiner reovirus, Grass carp reovirus, Striped bass reovirus and golden ide reovirus (genus Aquareovirus, family Reoviridae)

Full-length and partial genome sequences of four members of the genus Aquareovirus, family Reoviridae (Golden shiner reovirus, Grass carp reovirus, Striped bass reovirus and golden ide reovirus) were characterized. Based on sequence comparison, the unclassified Grass carp reovirus was shown to be a member of the species Aquareovirus C The status of golden ide reovirus, another unclassified aquareovirus, was also examined. Sequence analysis showed that it did not belong to the species Aquareovirus A or C, but assessment of its relationship to the species Aquareovirus B, D, E and F was hampered by the absence of genetic data from these species. In agreement with previous reports of ultrastructural resemblance between aquareoviruses and orthoreoviruses, genetic analysis revealed homology in the genes of the two groups. This homology concerned eight of the 11 segments of the aquareovirus genome (amino acid identity 17-42%), and similar genetic organization was observed in two other segments. The conserved terminal sequences in the genomes of members of the two groups were also similar. These data are undoubtedly an indication of the common evolutionary origin of these viruses. This clear genetic relatedness between members of distinct genera is unique within the family Reoviridae. Such a genetic relationship is usually observed between members of a single genus. However, the current taxonomic classification of aquareoviruses and orthoreoviruses in two different genera is supported by a number of characteristics, including their distinct G+C contents, unequal numbers of genome segments, absence of an antigenic relationship, different cytopathic effects and specific econiches.

Diving behaviour of freshwater finless porpoises (Neophocaena phocaenoides) in an oxbow of the Yangtze River, China

Dive-depth and swim-speed of a juvenile and an adult free-ranging, Yangtze finless porpoises (Neophocaena phocaenoides) were observed using velocity-time-depth recorders in an oxbow of the Yangtze River. In total, 8222 individual dives were recorded over 59 hours. Two dive types, deep-dive (greater than or equal to 2.7 m) and shallow-dive. were recognized. Horizontal travel distances of two finless pot-poises in a day were 94.4 km and 90.3 km, which were longer than those of oceanic relative species (harbor porpoises, Phocoena phocoena). Although the shallow water limited the maximum dive-depth, dive-duration, and bottom-time of finless porpoises were similar to the harbor porpoises. A sudden drop of swim-speed below 0.25 m s(-1) was frequently observed nearby the maximum dive-depth. This seemed to indicate "turning, around" behaviour, possibly during prey pursuit. (C) 2002 Published by Elsevier science Ltd on behalf of International Council for the Exploration of the Sea.

Floral surveys on Gomphonemaceae and Cymbellaceae (Bacillariophyta) from the headwaters of the Yangtze River, Qinghai, China

Gomphonemaceae and Cymbellaceae from the headwaters of the Yangtze River, Qinghai Province, China, comprised 84 taxa belonging to four genera. The dominant species were Gomphonema kaznakowi Mer., G. hedini Hust., G. olivaceum (Lyngbye) Kutz., Cymbella cistula (Ehr.) Kirchn. var. cistula and C. minuta Hilse ex Rabh. var. minuta. Some arctic and alpine forms also occurred, and the following taxa were unique to this region: C. cistula var. asiatica Mer., C. cistula var. capitata Grun., C. yabe Skvortzow var. punctata Li and Shi, G. olivaceum (Lyngbye) Kutzing var. brevistriatum Li and Shi and G. staurophorum (Pant.) Cleve-Euler var. oblongum Li and Shi. Different morphological forms of G. kaznakowi Mer. may be related to the upheaval of the plateau. Species diversity of the diatoms appears to be related not only to macro-environment (e.g., geographic zonation) but also to microhabitat and microclimate.

Retroposon analysis of major cetacean lineages: The monophyly of toothed whales and the paraphyly of river dolphins

SINE (short interspersed element) insertion analysis elucidates contentious aspects in the phylogeny of toothed whales and dolphins (Odontoceti), especially river dolphins. Here, we characterize 25 informative SINEs inserted into unique genomic loci during evolution of odontocetes to construct a cladogram. and determine a total of 2.8 kb per taxon of the flanking sequences of these SINE loci to estimate divergence times among lineages. We demonstrate that: (i) Odontocetes are monophyletic; (ii) Ganges River dolphins, beaked whales, and ocean dolphins diverged (in this order) after sperm whales; (iii) three other river dolphin taxa, namely the Amazon, La Plata, and Yangtze river dolphins, form a monophyletic group with Yangtze River dolphins being the most basal; and (iv) the rapid radiation of extant cetacean lineages occurred some 28-33 million years B.P., in strong accord with the fossil record. The combination of SINE and flanking sequence analysis suggests a topology and set of divergence times for odontocete relationships, offering alternative explanations for several long-standing problems in cetacean evolution.

Comparison between visual and passive acoustic detection of finless porpoises in the Yangtze river, China

Recently, sonar signals and other sounds produced by cetaceans have been used for acoustic detection of individuals and groups in the wild. However, the detection probability ascertained by concomitant visual survey has not been demonstrated extensively. The finless porpoises (Neophocaena phocaenoides) have narrow band and high-frequency sonar signals, which are distinctive from background noises. Underwater sound monitoring with hydrophones (B&K8103) placed along the sides of a research vessel, concurrent with visual observations was conducted in the Yangtze River from Wuhan to Poyang Lake in 1998 in China. The peak to peak detection threshold was set at 133 dB re 1 mu Pa. With this threshold level, porpoises could be detected reliably within 300 m of the hydrophone. In a total of 774-km cruise, 588 finless porpoises were sighted by visual observation and 44 864 ultrasonic pulses were recorded by the acoustical observation system. The acoustic monitoring system could detect the presence of the finless porpoises 82% of the time. A false alarm in the system occurred with a frequency of 0.9%. The high-frequency acoustical observation is suggested as an effective method for field surveys of small cetaceans, which produce high-frequency sonar signals. (C) 2001 Acoustical Society of America.

Sequence of genome segments 1, 2, and 3 of the grass carp reovirus (Genus Aquareovirus, family Reoviridae)

The genome segments 1, 2, and 3 of the grass carp reovirus (GCRV), a tentative species assigned to genus Aquareouirus, family Reouiridae, were sequenced. The respective segments 1, 2, and 3 were 3949, 3877, and 3702 nucleotides long. Conserved moths 5' (GUUAUUU) and 3' (UUCAUC) were found at the ends of each segment. Each segment contains a single ORF and the negative strand does not permit identification of consistent ORFs. Sequence analysis revealed that VP2 is the viral polymerase, while VPI might represent the viral guanyly/methyl transferase (involved in the capping process of RNA transcripts) and VP3 the NTPase/helicase (involved in the transcription and capping of viral RNAs), The highest amino acid identities (26-41%) were found with orthoreovirus proteins. Further genomic characterization should provide insight about the genetic relationships between GCRV, aquareoviruses, and orthoreoviruses, It should also permit to precise the taxonomic status of these different viruses. (C) 2000 Academic Press.

Occurrence of Bothriocephalus acheilognathi in cyprinid fish from three lakes in the flood plain of the Yangtze River, China

Cyprinid fish, Hemiculter leucisculus, Cultrichthys erythropterus and Culter dabryi, were sampled from Liangzi, Honghu and Tangxun lakes in the flood plain of the Yangtze River. The cestode Bothriocephalus acheilognathi Yamaguti, 1934 was found in the 3 lakes, but C. erythropterus sampled from Liangzi lake was found uninfected due probably to the small sample size. Findings of the cestode in the 3 lakes represent the first record of the parasite in the flood plain of the Yangtze River, indicating that B. acheilognathi may be distributed much wider in China than previously recognized.

The spatial pattern of the small fish community in the Biandantang Lake - a small shallow lake along the middle reach of the Yangtze River, China

The spatial pattern of the small fish community was studied seasonally in 1996 in the Biandantang Lake. Based on plant cover, the lake was divided into five habitats, arranged in the order by plant structure complexity from complex to simple: Vallisneria spiralis habitat (V habitat), Vallisneria spiralis-Myriophyllum spicatum habitat (V-M habitat), Myriophyllum spicatum habitat (M habitat), Nelunbo nucefera habitat (N habitat), and no vegetation habitat (NV habitat). A modified popnet was used for quantitative sampling of small fishes. A total of 16 fish species were collected; Hypseleotris swinhonis, Ctenogobius giurinus, Pseudorasbora parva, Carassius auratus and Paracheilognathus imberis were the five numerically dominant species. In both summer and autumn, the total density of small fishes was about 10 ind m(-2). Generally, Ctenogobius giurinus, a sedatory, benthic fish, was distributed more or less evenly among the five habitats, while the other four species had lower densities in the N habitat and NV habitat, which had the simplest structures. The distribution of the small fish species showed seasonal variations. In winter, most species concentrated in the V habitat, which had the most complex structure. In spring, the fish had low densities in the N and NV habitat, and were more or less evenly distributed in the other habitats. In summer, the fish had a low density in the NV habitat, and were evenly distributed in the other habitats. In autumn, the fish had higher densities in the V-M and M habitats than in the others. Generally, spatial overlaps between the dominant species were higher in winter than in the other seasons. It was suggested that the variations in the importance of predation risk and resource competition in habitat choice determined the seasonal changes of spatial patterns in the small fishes in the Biandantang Lake.

Effect of natural dissolved humic material on bioavailability and acute toxicity of fenpropathrin to the grass carp, Ctenopharyngodon idellus

The effects of aquatic humic acids on the bioconcentration and acute toxicity of fenpropathrin were evaluated using grass carp, Ctenopharyngodan idellus, in laboratory freshwater systems. The results demonstrated that both bioavailability and acute toxicity decreased in the presence of aquatic humic acid 5 and 10 mg/liter. In addition, the extent of influence increased with increasing concentration of aquatic humic acid, (C) 1999 Academic Press.