Polymorphic microsatellites in largemouth bronze gudgeon (Coreius guichenoti) developed from repeat-enriched libraries and cross-species amplifications

Largemouth bronze gudgeon (Coreius guichenoti) is a medium-sized fish endemic from the upper Yangtze River of China and its survival is threatened by the construction of the Three Gorges Dam. This study reports 20 new polymorphic microsatellites from a repeat-enriched genomic library with a mean number allele of 5.2, and observed and expected heterozygosities ranging from 0.035 to 1, and from 0.13 to 0.917, respectively. In a cross-species amplification test, nine of the 37 tested loci were found to be also polymorphic in a congeneric species, brass gudgeon (C. heterodon). In addition, other four loci from common carp (Cyprinus carpio) were also polymorphic in C. guichenoti. Out of these 24 polymorphic microsatellites, only three loci significantly deviated from Hardy-Weinberg equilibrium in the sampled population (P < 0.0025), and all pairwise tests for linkage disequilibrium among loci were nonsignificant after applying sequential Bonferroni correction (P > 0.0026). These novel microsatellites provide sufficient levels of polymorphism for studies on population genetics and conservation in C. guichenoti and its related species.

Redescription of Nais inflata Liang (Annelida : Oligochaeta) from China

Nais inflata Liang 1963 is redescribed on the basis of fully mature specimens collected from several localities of the Yangtze River and Yellow River. The observations and redescription resulting from the study of mature specimens supports N. inflata as a valid species. This species is characterized by a thickened body, the ventral crotchets all of the same type with distal tooth usually longer than proximal one, a greater number of dorsal chaetae per bundle, stout dorsal needles with equal minute bifid teeth (<1 mu m), presence of penial chaetae, and prostate glands on vasa deferentia instead of atria. The species is most closely related to N. communis Piguet, 1906 and N. variabilis Piguet, 1906. A table comparing allied species is provided.

Characterization of novel microsatellite loci in rare minnow (Gobiocypris rarus) and amplification in closely related species in Gobioninae

Rare minnow (Gobiocypris rarus) is an endangered small fish endemic to upper reach of the Yangtze River. From a (GT)n enriched genomic library, 32 microsatellites were isolated and characterized. Nineteen of these loci were polymorphic in a test population with alleles ranging from 2-7, and observed and expected heterozygosities from zero to 0.8438, and 0.2679 to 0.8264, respectively. In the cross-species amplifications, 13 out of 19 polymorphic loci were found to be also polymorphic in at least one of the 7 closely related species of the subfamily Gobioninae. These polymorphic microsatellite loci should provide sufficient level of genetic diversity to evaluate the fine-scale population structure in rare minnow and its closely related species for the conservation purpose.

Construction and characterization of a Lipotes vexillifer genomic DNA BAC library

We constructed a genomic DNA library for Lipotes vexillifer (L. vexillifer), the Baiji or Yangtze River dolphin, one of the most endangered mammals in the world. The library consists of 149,000 BAC clones, with an average insert size of 83 kb, representing approximately 3.4 haploid genome equivalents. PCR amplification of four known L. vexillifer genes yielded two to four positive clones each. To demonstrate the utility of this library, we isolated and sequenced the L. vexillifer alpha lactalbumin gene, which is a gene specific to mammals and one which has been widely used as molecular tool in phylogenetic analysis. We also end-sequenced 20 randomly selected clones, resulting in the identification of at least five new L. vexilliter genes, five SSR loci, and one SINE locus. These results suggest that this library is a valuable resource for candidate gene cloning, physical mapping, and genome sequencing of this important and threatened species.

Occurrence of Camallanus cotti in greatly diverse fish species from Danjiangkou Reservoir in central China

Two thousand four hundred fifty-eight fish comprised of 53 species were captured in the Danjiangkou Reservoir, in the northwestern part of Hubei Province, central China during 2004, to examine Camallanus cotti infections. We found that 19 cypriniform, 3 siluriforme, and 4 perciforme fishes were infected by the nematode. Our study revealed the species, Hemiculter bleekeri bleekeri, Culter oxycephaloide, Pseudolaubuca sinensis, Acanthobrama simony, Mylopharyngodon piceus, Ctenopharyngodon idella, Gnathopogon imberbis, G. argentatus, Saurogobio dabryi, S. dumerili, Gobiobotia ichangensis, Liobagrus marginatoides, and Ctenogobius shennongensis as new hosts of the worm. The number and range of fish host species found in this survey were much greater than any of the previous investigations. The mean prevalence, prevalence, mean abundance, and intensity of infection varied in different fish species, indicating a possible host preference. Moreover, we suggest that this nematode is a native parasite of cypriniform fishes in China, perhaps initially in the reaches of the Yangtze River.

Food composition and dietary overlap of macroinvertebrates in a shallow eutrophic lake in China: spatial and temporal variations

The present study was conducted in Lake Donghu, a suburban eutrophic lake arising from the middle reaches of the Yangtze River, China. Food composition of 32 taxa of zoobenthos was analyzed from 1251 gut samples. Macroinvertebrate primary consumers ingested mainly detritus, sand grains and diatoms. The predators primarily preyed on rotifers, crustaceans, oligochaetes and chironomid larvae. The dietary overlap was relatively high among collector taxa but low among other macroinvertebrates. Food composition and dietary overlap of macroinvertebrates changed considerably, both spatially and temporally. Food web structure differed between inshore and offshore regions of Lake Donghu. The inshore web was relatively complex and dynamic whereas the offshore web was simple and stable. Taxon-specific changes of diet seem to have little effect on the benthic food web structure in offshore waters of a eutrophic lake.

Distribution and morphology of two endemic gomphonemoid species, Gohphonema kaznakowi Mereschkowsky and G-yangtzensis Li nov sp in China

The morphology of Gomphonema kaznakowi Mereschkowsky was investigated using light microscopy. This species has two morphologically distinct areas near the headpole; an unornamented and an ornamented area. The two areas are distinguished from each other by the combination of size and striae number. A new species, Gomphonema yangtzensis Li nov. sp. is identified based on an ornamented area near the headpole. G. kaznakowi is reported from the upper and middle part of the Yangtze River, and was also found in the upper section of the Yellow River. G. yangtzensis was found in the upper area of the Yellow River and the middle of the Yangtze River. Their limited distribution may be due to certain environmental conditions or a different dispersal rate. Both species are illustrated.

Distribution of phytoplankton in the Three-Gorge Reservoir during rainy and dry seasons

After damming of the Yangtze River, in order to explore the impacts of the Three-Gorge Dam (TGD) on the aquatic ecosystem, phytoplankton composition, abundance and biomass spatial distribution were studied in the Three-Gorge Reservoir (TGR), and the closest upstream anabranch Xiangxi River, which is 38 kin away from the Three-Gorge Dam (TGD) during August (rainy season) 2004 and April (dry season) 2005. In surveys, 6 transects (2 downstream and 4 cross-stream) and 25 stations have been investigated and 314 samples were collected from the surface to the river bed with water samplers. In TGR, 63 taxa and 60 taxa were identified in the rainy and dry seasons, respectively. In the Xiangxi River, 39 taxa were observed in the rainy and dry seasons. Algal blooms occurred in the Xiangxi River and at the influx region of the Yangtze and Xiangxi in both seasons, but had not occurred prior to damming. In the rainy season, the dominant species was Chroomonas acuta with 1.84 x 10(7) cells l(-1), and in the dry season the dominant species were Asterionella formosa and Cryptomonas ovata with 1.34 x 10(7) cells l(-1) and 1.79 x 10(6) cells(.)l(-1), respectively. In the main channel of TGR, there were no significant correlations between phytoplankton abundance and the concentrations of the main soluble nutrients. In the Xiangxi River, significant negative correlations were observed between phytoplankton abundance and nitrate (Spearman, p < 0.01, n=21), phosphate (Spearman, p < 0.05, n=21) and silicate (Spearman, p < 0.01, n=21) in the rainy season, and similar correlations were also observed with nitrate (Spearman, p < 0.05, n=28) and silicate (Speannan, p < 0.01, n=28), but not with phosphate in the dry season. Since the damming of the Yangtze River, eutrophication in the anabranch within the backwater has occurred and become severe, and the frequency of algal bloom within TGR and anabranches is expected to increase. (c) 2006 Elsevier B.V. All rights reserved.

Biological mechanisms driving the seasonal changes in the internal loading of phosphorus in shallow lakes

Because of the obvious importance of P as a nutrient that often accelerates growth of phytoplankton (including toxic cyanobacteria) and therefore worsens water quality, much interest has been devoted to P exchange across the sediment-water interface. Generally, the release mode of P from the sediment differed greatly between shallow and deep lakes, and much of the effort has been focused on iron and oxygen, and also on the relevant environmental factors, for example, turbulence and decomposition, but a large part of the P variation in shallow lakes remains unexplained. This paper reviews experimental and field studies on the mechanisms of P release from the sediment in the shallow temperate (in Europe) and subtropical (in the middle and lower reaches of the Yangtze River in China) lakes, and it is suggested that pH rather than DO might be more important in driving the seasonal dynamics of internal P loading in these shallow lakes, i.e., intense photosynthesis of phytoplankton increases pH of the lake water and thus may increase pH of the surface sediment, leading to enhanced release of P (especially iron-bound P) from the sediment. Based on the selective pump of P (but not N) from the sediment by algal blooms, it is concluded that photosynthesis which is closely related to eutrophication level is the driving force for the seasonal variation of internal P loading in shallow lakes. This is a new finding. Additionally, the selective pump of P from the sediment by algal blooms not only explains satisfactorily why both TP and PO4-P in the hypereutrophic Lake Donghu declined significantly since the mid-1980s when heavy cyanobacterial blooms were eliminated by the nontraditional biomanipulation (massive stocking of the filter-feeding silver and bighead carps), but also explains why TP in European lakes decreased remarkably in the spring clear-water phase with less phytoplankton during the seasonal succession of aquatic communities or when phytoplankton biomass was decreased by traditional biomanipulation. Compared with deep lakes, wax and wane of phytoplankton due to alternations in the ecosystem structure is also able to exert significant influences on the P exchange at the sediment-water interface in shallow lakes. In other words, biological activities are also able to drive P release from sediments, and such a static P release process is especially more prominent in eutrophic shallow lakes with dense phytoplankton.

Biogeography and molecular phylogeny of the genus Schizothorax (Teleostei : Cyprinidae) in China inferred from cytochrome b sequences

Aim: To test a vicariant speciation hypothesis derived from geological evidence of large-scale changes in drainage patterns in the late Miocene that affected the drainages in the south-eastern Tibetan Plateau. Location: The Tibetan Plateau and adjacent areas. Methods: The cytochrome b DNA sequences of 30 species of the genus Schizothorax from nine different river systems were analysed. These DNA sequences were analysed using parsimony, maximum likelihood and Bayesian methods. The approximately unbiased and Shimodaira-Hasegawa tests were applied to evaluate the statistical significance of the shortest trees relative to alternative hypotheses. Dates of divergences between lineages were estimated using the nonparametric rate smoothing method, and confidence intervals of dates were obtained by parametric bootstrapping. Results: The phylogenetic relationships recovered from molecular data were inconsistent with traditional taxonomy, but apparently reflected geographical associations with rivers. Within the genus Schizothorax, we observed a divergence between the lineages from the Irrawaddy-Lhuit and Tsangpo-Parlung rivers, and tentatively dated this vicariant event back to the late Miocene (7.3-6.8 Ma). We also observed approximately simultaneous geographical splits within drainages of the south-eastern Tibetan Plateau, the Irrawaddy, the Yangtze and the Mekong-Salween rivers in the late Miocene (7.1-6.2 Ma). Main conclusions: Our molecular evidence tentatively highlights the importance of palaeoriver connections and the uplift of the Tibetan Plateau in understanding the evolution of the genus Schizothorax. Molecular estimates of divergence times allowed us to date these vicariant scenarios back to the late Miocene, which agrees with geological suggestions for the separation of these drainages caused by tectonic uplift in south-eastern Tibet. Our results indicated the substantial role of vicariant-based speciation in shaping the current distribution pattern of the genus Schizothorax.

Genetic structure and low-genetic diversity suggesting the necessity for conservation of the Chinese Longsnout catfish, Leiocassis longirostris (Pisces : Bagriidae)

Genetic variation and phylogenetic relationship of Leiocassis longirostris populations from the Yangtze River were investigated at mitochondrial DNA level. The samples were collected from the upstream and mid-downstream of the Yangtze River. Three mitochondrial DNA fragments, ND5/6, cytochrome b (Cyt b) and control region (D-loop), were amplified and then digested by 10 restriction endonucleases. Twenty-three D-loop fragments randomly selected were sequenced. Digestion patterns of ND5/6 by AluI and HaeIII, D-loop by HinfI and RsaI, and Cyt b by HaeIII were polymorphic. Ten and eighteen haplotypes were obtained from RFLP data and sequence data, respectively. The individuals from upstream and mid-downstream of the Yangtze River were apparently divided into two groups. The average genetic distance was 0.008 and 0.010 according to the two data. Low diversities and decreasing abundance indicated that Leiocassis longirostris may be in severe danger and reasonable measures of fishery management should be taken.

First description of the male and redescription of the female of Philometra clavaeceps Dogiel and Akhmerov, 1959 (Nematoda : Philometridae), a parasite of east Asian cyprinid fish

The first description of the male and a redescription of the female of the nematode Philometra clavaeceps Dogiel and Akhmerov, 1959, a parasite of east Asian cyprinids, are presented on the basis of specimens collected from Culter erythropterus Basilewsky and Culler dabryi Bleeker from Liangzi Lake (the Yangtze River basin), Hubei Province, central China. Gravid females from the fish abdominal cavity, penetrating often into ovaries, occurred in May-June, whereas conspecific males and young mature females on the swimbladder were recorded in January. Philometra clavaeceps seems to have a pronounced annual maturation cycle in the locality. The finding of P. clavaeceps in C. dabryi represents a new host record.

Isolation and characterization of microsatellites in Chinese sturgeon, Acipenser sinensis

From (GATA)(n) and (AAAG)(n) enriched genomic libraries for the Chinese sturgeon (Acipenser sinensis), 50 primer pairs were developed using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Forty-six primer pairs exhibited highly polymorphic with two to 11 alleles per locus, while the rest four displayed monomorphic. These markers yielded 246 alleles in a survey of eight specimens of wild A. sinensis. Average observed heterozygosity ranged from 0.13 to 1.00. These loci should provide sufficient levels of genetic diversity to allow parentage analysis for artificial stocking management and delineation of fine-scale population structure.

Distribution, feeding and body condition of four small fish species in the near-shore and central areas of Liangzi Lake, China

Small fish abundance is usually high in heavily vegetated habitats in Yangtze lakes, China. Visual and swimming barriers created by dense macrophytes beds could reduce feeding efficiency and growth of small fishes. We tested the hypothesis that small fishes in habitats with dense macrophytes would show decreased feeding efficiency and reduced growth rates by comparing feeding efficiency (measured as the relative weight of fore-gut contents), total length, and condition factor of four small young-of-the-year fishes collected in the near-shore (heavily vegetated) and central (less vegetated) areas of Liangzi Lake. Feeding efficiency, total length, or condition factor were each significantly reduced in the near-shore area compared with the central area for Ctenogobius giurinus, Pseudorasbora parva and Carassius auratus auratus. This supports our hypothesis that vegetation abundance may mediate feeding efficiency and growth of small fishes. Although Hypseleotris swinhonis did not show significant decreases in feeding efficiency or growth in the near-shore area, there was not any reversed tendency, i.e. increased feeding rate or growth in the near-shore area compared to the central area.

Seasonal dynamics and distribution of the digenean Phyllodistomum pawlovskii (Trematoda : Gorgoderidae) in the bullhead catfish, Pseudobagrus fulvidraco, in a lake of China

Seasonal population dynamics of the digenean Phyllodistomum pawlovskii in the urinary bladder of the bullhead catfish, Pseudobagrus fulvidraco, were investigated in Liangzi Lake in the flood plain of the Yangtze River in China from February 2001 to July 2002. The overall prevalence of the parasite was high, 41.5% (n = 1,476), while the mean abundance was relatively low, 1.24 +/- 2.11. The parasite exhibited evident seasonality in changes of prevalence and abundance. In brief, prevalence and abundance were very low in midwinter (January), but increased and remained relatively high in other seasons and months. The distribution pattern of this parasite in the fish was overdispersed, with a variance to mean ratio > 1, but its frequency distribution could not be described by the negative binomial model. There were positive correlations between the number of the parasites per fish and the age and length of the fish; a peaked age-parasite abundance curve was not detected in the parasite-host association. It is suggested that the parasite P. pawlovskii has little effect on the population structure of the bullhead catfish.