Macrozoobenthic community of Fuxian Lake, the deepest lake of southwest China

Located in the Yunnan-Guizhou Plateau in southwest China, Fuxian Lake covers an area of 211 km(2), with maximum depth of 155 m. It is known to have a unique fauna, including 14 described endemic species. In order to describe the zoobenthic community of the lake more completely, the present study was conducted from August 2002 to August 2003. Altogether 62 benthic taxa, including 22 oligochaetes, 21 molluscs and 18 insects were identified, of which the dominant taxa belonged to Potamothrix, Procladius and Paraprososthenia. The standing stocks of benthos were much higher in the littoral (824 ind/m(2) in density, 3.72 g/m(2) in biomass) than in the profundal region (23 ind/m(2) in density, 0.10 g/m(2) in biomass). Species richness was greatest in summer and standing stocks were larger in spring and summer than in other seasons. Analyses of functional feeding groups indicated that collector-gatherers and scrapers were predominant in entire lake. Stepwise multiple regression analysis demonstrated that the water depth is the most important factor affecting the distribution of macrozoobenthos. (C) 2008 Elsevier GmbH. All rights reserved.

Threatened fishes of the world: Tanichthys albonubes Lin 1932 (Cyprinidae)

The white cloud mountain minnow Tanichthys albonubes Lin is an endemic species to southern China and the genus has two species, Tanichthys albonubes Lin and Tanichthys micagemmae Freyhof et Herder. The distribution range of T. albonubes Lin and T. micagemmae is very narrow and only found in the mountain brooks of Baiyunshan Mountain (White Cloud Mountain), Huaxian Country and the vicinity of Guangzhou in Guangdong Province and Halong, Quang Ninh Province, Vietnam respectively. The wild populations of this fish had already been on the verge of extinction when Shu-Yan Lin first discovered it in 1932 at the Baiyunshan Mountain. It was believed to be extinct in the wild because there were no reports of this fish in the wild since 1980. In September 2003, a small and isolated population of the fish was discovered in a mountain puddle in the north vicinity of Guangzhou. Additional studies are needed to determine the survival and propagation of the released fish. The protection of their natural habitat should be implemented.

Description of a new genus and two new species of labeonine fishes from South China (Teleostei : Cyprinidae)

A new genus and two new species are described from the Pearl River drainage in Guangxi Province, South China. Hongshuia, new genus, can be distinguished from all other Asian genera of the Labeonini by having a lower lip with its median lobe modified into a round, fleshy plate peripherally greatly protruded so as to form a ring-like fold that is posteromedially continuous with the mental region, and centrally sunken so as to form a round, flat, fleshy pad. This genus is distinct from all other Asian labeonine genera of the Garrina except for one newly described species of Parasinilabeo ( P. longibarbus), Pseudocrossocheilus, and Sinocrossocheilus, in the presence of well-developed maxillary barbels. Hongshuia differs from the above three genera in the lower lip morphology, and further from both Pseudocrossocheilus and Qianlabeo in the number of pharyngeal tooth rows and from Sinocrossocheilus in the colour pattern. Two new species, H. banmo and H. paoli, differ in the distribution density and degree of development of papillae on the rostral fold, depth of indentations on the distal edge of the rostral fold, presence or absence of papillae on the lower lip, size and shape of tubercles on the tip of the snout and anterior portion of the lachrymal, length, position and colour pattern of the dorsal fin, and snout length.

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.

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.

Molecular evidence for the monophyly of East Asian groups of Cyprinidae (Teleostei : Cypriniformes) derived from the nuclear recombination activating gene 2 sequences

The family Cyprinidae is one of the largest families of fishes in the world and a well-known component of the East Asian freshwater fish fauna. However, the phylogenetic relationships among cyprinids are still poorly understood despite much effort paid on the cyprinid molecular phylogenetics. Original nucleotide sequence data of the nuclear recombination activating gene 2 were collected from 109 cyprinid species and four non-cyprinid cypriniform outgroup taxa and used to infer the cyprinid phylogenetic relationships and to estimate node divergence times. Phylogenetic reconstructions using maximum parsimony, maximum likelihood, and Bayesian analysis retrieved the same clades, only branching order within these clades varied slightly between trees. Although the morphological diversity is remarkable, the endemic cyprinid taxa in East Asia emerged as a monophyletic clade referred to as Xenocypridini. The monophyly for the subfamilies including Cyprininae and Leuciscinae, as well as the tribes including Labeonini, Gobionini, Acheilognathini, and Leuciscini, was also well resolved with high nodal support. Analysis of the RAG2 gene supported the following cyprinid molecular phylogeny: the Danioninae is the most basal subfamily within the family Cyprinidae and the Cyprininae is the sister group of the Leuciscinae. The divergence times were estimated for the nodes corresponding to the principal clades within the Cyprinidae. The family Cyprinidae appears to have originated in the mid-Eocene in Asia, with the cladogenic event of the key basal group Danioninae occurring in the early Oligocene (about 31-30 MYA), and the origins of the two subfamilies, Cyprininae and Leuciscinae, occurring in the mid-Oligocene (around 26 MYA). (c) 2006 Elsevier Inc. All rights reserved.

Insecticide resistance in Chinese populations of the Culex pipiens complex through esterase overproduction

In most parts of China, mosquitoes have been subjected to organophosphate (OP) insecticide treatments since the mid-1960s, and resistance gene monitoring in the Culex pipiens complex (Diptera: Culicidae) started in only a few locations from the end of the 1980s. Many resistant alleles at the Ester locus have been found in field populations, including those commonly found around the world (Ester(B1) and Ester(2)), and those endemic to China (Ester(B6), Ester(B7), Ester(8), and Ester(9)). This situation is atypical, and may represent a complex situation for the evolution of insecticide resistance genes in China. To increase our understanding of the Chinese situation and our ability to manage resistance in the C. pipiens complex, a large study was performed. Twenty field populations were sampled from Beijing to Guangzhou. Bioassays with five insecticides (dichlorvos, parathion, chlorpyrifos, 2-sec-butylphenyl methyl carbamate, and propoxur) disclosed resistance levels variable according to the geographic origin, and up to 85-fold for dichlorvos. Six overproduced esterases were identified, including two that have not been previously described. Most of them were found in all samples, although at variable frequencies, suggesting variable selection or a transient situation, e.g., each one was recently restricted to a particular geographic area. The results are discussed in the context of recent alterations to insecticide campaigns, and of the evolution of resistance genes in Chinese C. pipiens populations.

Molecular phylogenetics of Gymnocypris (Teleostei : Cyprinidae) in Lake Qinghai and adjacent drainages

149 complete mitochondrial DNA (mtDNA) cytochrome b (Cyt b) genes (1140 bp) of Gymnocypris przewalskii, Gymnocypris eckloni and Gymnocyptis scolistomus from the Lake Qinghai, Yellow River and Qaidam Basin were sequenced and analyzed. Consistent dendrogram indicated that the samples collected from the same species do not constitute a separate monophyletic group and all the samples were grouped into three highly divergent lineages (A, B and C). Among them, Lineage A contained all samples of G. przewalskii from the Lake Qinghai and partial samples of the G. eckloni from the Yellow River. Lineage B contained the remaining samples of G. eckloni from the Yellow River. Lineage C was composed of a monophyletic group by G. eckloni from the Qaidam Basin. Analysis of molecular variance (AMOVA) indicated that most of genetic variations were detected within these three mtDNA lineages (93.12%), suggesting that there are three different lineages of Gymnocypris in this region. Our Cyt b sequence data showed that G. przewalskii was not a polytypic species, and G. scolistomus was neither an independent species nor a subspecies of G. eckloni. The divergent mtDNA lineages of G. eckloni from the Yellow River suggested that gene flow between the different populations was restricted to a certain extent by several gorges on the upper reach of the Yellow River. Lineage B of G. eckloni might be the genetic effect from the ancestor which was incorporated with the endemic schizothoracine fishes when the headward erosion of the Yellow River reached to its current headwaters of late. The G. eckloni from Basin Qaidam was a monophyletic group (lineage C) and F-st values within G. eckloni from the Yellow River were higher than 0.98, suggesting that the gene flow has been interrupted for a long time and the G. eckloni from Basin Qaidam might have been evolved into different species by ecology segregation. The correlation between the rakers number of Gymnocypris and population genetic variation was not significant. All Gymnocypris populations exhibited a low nucleotide diversity (pi = 0.00096-0.00485). Therefore the Gymnocyptis populations from Basin Qaidam could have experienced severe bottleneck effect in history. Our result suggested Gym-nocypris populations of Basin Qaidam should give a high priority in conservation programs.

Population genetic structure of the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis): Implications for management and conservation

Understanding the population genetic structure is a prerequisite for conservation of a species. The degree of genetic variability characteristic of the mitochondrial DNA control region has been widely exploited in studies of population genetic structure and can be useful in identifying meaningful population subdivisions. To estimate the genetic profile of the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis), an endangered freshwater population endemic to China, the complete mtDNA control region was examined in 39 individuals belonging to seven different stocks inhabiting the middle and lower reaches of the Yangtze River. Very low genetic diversity was found (nucleotide diversity 0.0011 +/- 0.0002 and haplotypic diversity 0.65 +/- 0.05). The mtDNA genetic pattern of the Yangtze population appears to indicate a founder event in its evolutionary history and to support the marine origin for this population. Analyses by F-st and Phi(st) yielded statistically significant population genetic structure (F-st = 0.44, P < 0.05; phi(st) = 0.36, P < 0.05). These results may have significant implications for the management and conservation of the Yangtze finless porpoise in the future.

Revision of the genus Niwaella in China (Pisces, Cobitidae), with description of two new species

Loaches of the genus Niwaella, family Cobitidae, are distributed only in East Asia. At present only in Japan and South Korea have fishes of the genus Niwaella been found. Herein we revise the genus Niwaella in China. Son and He ( 2001) transferred the species Cobitis laterimaculata to the genus Niwaella, but their specimens were not N. laterimaculata, but a new species, N. longibarba sp. n., collected from Cao'ejiang River, Huangzezhen, Chengxian County, Zhejiang Province. The new species is distinguished from N. laterimaculata by its colour pattern of a row of slightly large, and long, scattered dark brown vertical bars on the dorsolateral surface, two or three striations on the caudal fin, and long barbels and undeveloped mental lobes. In this paper we also describe another new species, N. xinjiangensis sp. n., collected from Xinjiang River, Guangfeng County, Jiangxi Province, May 1990, with diagnostic colour pattern of 17 - 20 large and long, dark brown vertical bars on the dorsolateral surface, a dark stripe or rounded black spots along the lateral midline and some blotches below the lateral midline; it is a large-sized species, with shorter barbels, and longer caudal peduncle. Thus five species of the genus Niwaella are known, three are endemic to eastern China and two are endemic to either Japan or South Korea.

Morphology, biometry and distribution of Difflugia biwae Kawamura, 1918 (Protozoa : Rhizopoda)

The freshwater testate amoeba Difflugia biwae Kawamura, 1918, isolated from Mulan Lake, Hubei Province, China is investigated using light microscopy and scanning electron microscopy. The morphology, biometry and distribution of this little known species are supplied. After careful comparison with three other similar species, including D. delicatula Gauthier-Livre et Thomas, D. elegans Penard and D. oblonga caudata Stepanek, we believe that the characteristics of smooth fusiform shell, conspicuous great collar flare (always larger than body-width) around the aperture, constriction behind the aperture and a somewhat curved aboral horn set D. biwae apart from all other Difflugia species. This species shows a great diversity in total length, collar height and aboral horn length which have high variability (CV between 11.76 and 24.52). However, body width, collar diameter, neck width, body length and aperture diameter are fairly constant with low variability (CV between 5.34 and 8.79) which shows a remarkable uniformity of D. biwae. Also, the size frequency distributions of both body width and body length yield bell-shaped (normally distributed) curves and indicate that D. biwae is a size-monomorphic species, characterized by a main-size class and a small size range. D. biwae is probably endemic to East Asia (China and Japan) because it has such a large size (165-306 mu m) that it would have been easily found in Europe and North America, if it were there. Consequently, D. biwae must have a restricted geographical distribution, disproving the old hypothesis that microscopic organisms are cosmopolitan.

Molecular phylogeny of three subspecies of common carp Cyprinus carpio, based on sequence analysis of cytochrome b and control region of mtDNA

The complete cytochrome b and the control region of mtDNA (about 2070 bp in total) of 10 strains belonging to three subspecies of the common carp, including three wild subspecies (the Yangtze River wild common carp - Cyprinus carpio haematopterus, Yuanjiang River wild common carp Cyprinus carpio rubrofuscus and Volga River wild common carp - Cyprinus carpio carpio) and seven domestic strains (Xingguo red carp, Russian scattered scaled mirror carp, Qingtian carp, Japanese Koi carp, purse red carp, Big-belly carp, German mirror carp) were sequenced. Phylogenetic analysis indicated that the 10 strains form three distinct clades, corresponding to C. c. haematopterus, C. c. rubrofuscus and C. c. carpio respectively. Purse red carp, an endemic domestic strain in Jiangxi province of China, showed a higher evolution rate in comparison with the other strains of C. c. haematopterus, most probably because of intensive selection and a long history of domestication. Base variation ratios among the three subspecies varied from 0.78% (between C. c. haematopterus and C. c. rubrofuscus) to 1.47%(between C. c. carpio and C. c. rubrofuscus). The topography of the phylogenetic tree and the geographic distribution of three subspecies closely resemble each other. The divergence time between C. c. carpio and the other two subspecies was estimated to be about 0.9 Myr and about 0.5 Myr between C. c. haematopterus and C. c. rubrofuscus. Based on phylogenetic analysis, C. c. rubrofuscus might have diverged from C. c. haematopterus.

Spawning areas and early development of long spiky-head carp (Luciobrama macrocephalus) in the Yangtze River and Pearl River, China

The spawning areas and early development of long spiky-head carp, Luciobrama macrocephalus (Lacepede), an endemic fish species in China, were investigated in the Yangtze River and Pearl River of central and southeastern China between 1961 and 1993. The potamodromous fish migrated upstream to spawn between May and July as the floodwater began to rise. The water-hardened eggs drifted down the river, and the embryos and larvae developed in the course of drifting. The spawning areas of the fish were widely found in the upper and middle main channels and large tributaries. Two large dams (Gezhouba dam and Danjiangkou dam) did not significantly impact on the reproduction of the fish. Fifty stages of the early development from one cell to the juvenile with fully formed fins were observed and characterized pictorially. The larvae of long spiky-head carp could be distinguished from the larvae of other co-occurring species by counting the number of somites and comparing the proportion of sizes of eye to otic capsule.

Experimental and field studies of Mesocyclops notius (Copepoda : Cyclopoida) from a subtropical Chinese lake, Lake Donghu

This paper reports experimental and field studies on the cyclopoid Mesocyclops notius from subtropical Lake Donghu close to the Yangtze river. Mesocyclops notius, a dominant crustacean Zooplankter throughout tropical Australia, was previously considered to be endemic to Australia, but recently, Mesocyclops leuckarti in Lake Donghu was re-identified as M. notius. Laboratory culture experiments were conducted to reveal the effect of temperature (15, 20, 25 and 30 degreesC) on the development, growth and reproduction of M. notius. Temperature was inversely related to development times of eggs, nauplii and copepodites, body length and physiological longevity of adults, and brood size. Body length and physiological longevity of females were greater than those of males at the same temperature. No reproduction took place at 15 degreesC. Production and seasonal cycles of M. notius during 1980-1982 were studied at two sampling stations of Lake Donghu. At the mid-lake station, the annual production and production/biomass (P/B) ratio of M. notius varied between 6.0 and 18.0 g dry wt m(-2) year(-1) and 74.6 and 95.5, respectively. Mesocyclops notius reached their highest density peaks in the warm months (July-October), with a maximum density of 1256 individuals l(-1) at a littoral site. No reproduction and recruitment by AL notius took place during the cold months (December March) when the temperature of the lake water was < 15 degreesC. Mesocyclops notius were more abundant at a littoral station than at a pelagic station, possibly due to different food availability. The higher male:female sex ratio of M. notius at the littoral station was most likely caused by size-selective fish predation on larger females.

Molecular phylogenetic systematics of twelve species of acipenseriformes based on mtDNA ND4L-ND4 gene sequence analysis

Acipenseriformes is an endangered primitive fish group, which occupies a special place in the history of ideas concerning fish evolution, even in vertebrate evolution. However, the classification and evolution of the fishes have been debated. The mitochondrial DNA (mtDNA) ND4L and partial ND4 genes were first sequenced in twelve species of the order Acipenseriformes, including endemic Chinese species. The following points were drawn from DNA sequences analysis: (i) the two species of Huso can be ascribed to Acipenser; (ii) A. dabryanus is the mostly closely related to A. sinensis, and most likely the landlocked form of A. sinensis; (iii) genus Acipenser in trans-Pacific region might have a common origin; (iv) mtDNA ND4L and ND4 genes are the ideal genetic markers for phylogenetic analysis of the order Acipenseriformes.