Molecular phylogenetics of the family Cyprinidae (Actinopterygii : Cypriniformes) as evidenced by sequence variation in the first intron of S7 ribosomal protein-coding gene: Further evidence from a nuclear gene of the systematic chaos in the family

The family Cyprinidae is the largest freshwater fish group in the world, including over 200 genera and 2100 species. The phylogenetic relationships of major clades within this family are simply poorly understood, largely because of the overwhelming diversity of the group; however, several investigators have advanced different hypotheses of relationships that pre- and post-date the use of shared-derived characters as advocated through phylogenetic systematics. As expected, most previous investigations used morphological characters. Recently, mitochondrial DNA (mtDNA) sequences and combined morphological and mtDNA investigations have been used to explore and advance our understanding of species relationships and test monophyletic groupings. Limitations of these studies include limited taxon sampling and a strict reliance upon maternally inherited mtDNA variation. The present study is the first endeavor to recover the phylogenetic relationships of the 12 previously recognized monophyletic subfamilies within the Cyprinidae using newly sequenced nuclear DNA (nDNA) for over 50 species representing members of the different previously hypothesized subfamily and family groupings within the Cyprinidae and from other cypriniform families as outgroup taxa. Hypothesized phylogenetic relationships are constructed using maximum parsimony and Basyesian analyses of 1042 sites, of which 971 sites were variable and 790 were phylogenetically informative. Using other appropriate cypriniform taxa of the families Catostomidae (Myxocyprinus asiaticus), Gyrinocheilidae (Gyrinocheilus aymonieri), and Balitoridae (Nemacheilus sp. and Beaufortia kweichotvensis) as outgroups, the Cyprinidae is resolved as a monophyletic group. Within the family the genera Raiamas, Barilius, Danio, and Rasbora, representing many of the tropical cyprinids, represent basal members of the family. All other species can be classified into variably supported and resolved monophyletic lineages, depending upon analysis, that are consistent with or correspond to Barbini and Leuciscini. The Barbini includes taxa traditionally aligned with the subfamily Cyprininae sensu previous morphological revisionary studies by Howes (Barbinae, Labeoninae, Cyprininae and Schizothoracinae). The Leuciscini includes six other subfamilies that are mainly divided into three separate lineages. The relationships among genera and subfamilies are discussed as well as the possible origins of major lineages. (c) 2008 Published by Elsevier Inc.

Triplophysa lixianensis, a new nemacheiline loach species (Pisces : Balitoridae) from the upper Yangtze River drainage in Sichuan Province, South China

Triplophysa lixianensis, a new nemacheiline loach species, is described from the Min Jiang of the upper Yangtze River drainage in Sichuan Province, South China. It can be separated from all other species of Triplophysa by having a unique combination of the following characters: posterior chamber of gas bladder greatly reduced or absent; caudal peduncle columnar with a roughly round cross- section at its beginning; anterior edge of lower jaw completely exposed or uncovered by lower lip; intestine short, forming a zigzag loop below stomach; dorsal- fin origin closer to caudal- fin base than to snout tip; pelvic fin inserted anterior to dorsal- fin origin; snout length 50.6 - 57.5 % of head length; eye diameter 12.3 15.4 % of head length; caudal peduncle length 25.1 - 27.1 % of standard length; anal fin with five branched rays; lower lip greatly furrowed with two thick lateral lobes; and body smooth or scaleless.

A new species of Allocreadium (Trematoda : Allocreadiidae) from freshwater fishes in the Danjiangkou Reservoir in China

A new species of Allocreadium, Allocreadium danjiangensis n. sp., is described from the intestine of several species of freshwater fish, including Abbottina rivularis (Basilewsky, 1855), Sarcocheilichthys nigripinnis nigripinns (Gunther, 1873), Gnathopogon argentatus (Sauvage et Dabry 1874), Opsariichthys uncirostris bidens (Gunther, 1873), and Erythroculter mongolicus mongolicus (Basilewsky, 1855) (Cyprinidae) from the Danjiangkou Reservoir in central China. The main morphological characters of the new species are as follows: vitelline follicles numerous, extending from the level of acetabulum to posterior extremity, distributed over both sides around the ceca; cirrus sac relatively large, developed, lying obliquely anterior to the acetabulum, extending from the level of the intestinal bifurcation to the central level of acetabulum, and overlapping left or right cecal; and ovary much smaller than testes, generally close to or even overlapping the anterior border of anterior testis. Observation by scanning electron microscopy shows only 2 kinds of tegumental formations, i.e., papillae and tubercles, instead of 3 types of tegumental formations, i.e., papillae, bosses, and minute sensor receptors observed on other species of the Allocreadiidae. The tegumental striations of the present species vary on the different parts of the body. In addition, a new structure, identified as the "groove" with a tonguelike tubercle, was observed on the inner wall of acetabulum.

Morphological variability between wild populations and inbred stocks of a Chinese minnow, Gobiocypris rarus

Gobiocypris rarus, a small, native cyprinid fish, is currently widely used in research on fish pathology, genetics, toxicology, embryology, and physiology in China. To develop this species as a model laboratory animal, inbred strains have been successfully created. In this study, to explore a method to discriminate inbred strains and evaluate inbreeding effects, morphological variation among three wild populations and three inbred stocks of G. rarus was investigated by the multivariate analysis of eight meristic and 30 morphometric characters. Tiny intraspecific variations in meristic characters were found, but these were not effective for population distinction. Stepwise discriminant analysis and cluster analysis of conventional measures and truss network data showed considerabe divergence among populations, especially between wild populations and inbred stocks. The average discriminant accuracy for all populations was 82.1% based on conventional measures and 86.4% based on truss data, whereas the discriminant accuracy for inbred strains was much higher. These results suggested that multivariate analyses of morphometric characters are an effective method for discriminating inbred strains of G. rarus. Morphological differences between wild populations and inbred strains appear to result from both genetic differences and environmental factors. Thirteen characters, extracted from stepwise discriminant analysis, played important roles in morphological differentiation. These characters were mainly measures related to body depth and head size.

Bibarba bibarba: A new genus and species of Cobitinae (Pisces : Cypriniformes : Cobitidae) from Guangxi Province (China)

A new genus of Cobitinae, Bibarba gen. n., and a new species, B. bibarba sp. n., were discovered and are described for the Chengjiang River, a tributary of the Hongshuihe River in Guangxi Province of southern China. This river region is characterized by a Karst landscape, and the river that is inhabited by the new genus is a slowly moving stream with arenaceous and cobblestone beds. The new genus resembles Cobitis Linnaeus, 1758 (subfamily Cobitinae) in the shape and pigmentation pattern of their body, the absence of scales on their head, and the presence of a suborbital spine, but differs from it by a single Lamina circularis on the third pectoral fin ray instead of on the base of the second pectoral fin ray; two pairs of barbels (one rostral pair and one maxillo-mandibular pair) instead of three pairs of barbels (one rostral pair, one maxillary pair, and one maxillo-mandibular pair); a relatively thick and short suborbital spine with a strong medio-lateral process instead of a suborbital spine without or with a weakly formed medio-lateral process as in Cobitis; and the lack of a black stripe extending from the occiput through the eye to the insertion of the rostral barbel. The first two characters have not been reported in any other genus of the subfamily Cobitinae. A morphometric character analysis based on PCA reveals differences between B. bibarba and C. sinensis in body size, barbel length, interorbital width, pectoral fin length in males, and the position of the dorsal and ventral fins. Type specimens of the new species are kept in the Freshwater Fishes Museum of the Institute of Hydrobiology at the Chinese Academy of Sciences in Wuhan, Hubei Province. (c) 2007 Elsevier GmbH. All rights reserved.

Morphological and genetic differences of Trypanosoma in some Chinese freshwater fishes: difficulties of species identification

Blood smears and purified trypanosome from freshwater fishes yellow catfish (Pseudobagras fulvidraco) and common carp (Cyprinus carpio) captured from Niushan Lake, Hubei Province were examined to determine whether all of their trypanosomes were Trypanosoma pseudobagri, a species of supposed host specificity and widespread existence across China. Trypanosomes occurred in 16/16 blood smears, and morphometric character analysis of trypanosomes from these smears showed that there were three morphospecies, Trypanosoma sp Carpio, T. sp Pseudobagri, and T. sp. 18S rDNA sequences of trypanosomes from 16 samples revealed three genetic groups among these fish trypanosomes. Group 1 was from C. carpio containing T. sp Carpio; groups 2 and 3 were from P. fulvidraco containing T. sp Pseudobagri and T. sp, respectively. The high similarity of morphometric characters and 18S rDNA sequences showed that T. sp Carpio and T. siniperca probably were the same species. T. sp Pseudobagri was the first occurrence in China. Sequence comparison showed that T. sp Pseudobagri sequence was most similar to that of clone Marv, whereas T. sp sequence differ from those of T. sp Carpio and T. sp Pseudobagri by 5.4 and 5.8%, respectively, and tentatively identified as T. pseudobagri. It was concluded that three species of trypanosomes, at least three genotypes occur in Niushan Lake fishes, and P. fulvidraco in this region appear to contain both types, although the identification of T. pseudobagri remains a problem.

Short and long peptidoglycan recognition proteins (PGRPs) in zebrafish, with findings of multiple PGRP homologs in teleost fish

Peptidoglycan recognition protein (PGRP) specifically binds to peptidoglycan and is considered to be one of the pattern recognition proteins in the innate immunity of insect and mammals. Using a database mining approach and RT-PCR, multiple peptidoglycan recognition protein (PGRP) like genes have been discovered in fish including zebrafish Danio rerio, Japanese pufferfish TakiFugu rubripes and spotted green pufferfish Tetraodon nigroviridis. They share the common features of those PGRPs in arthropod and mammals, by containing a conserved PGRP domain. Based on the predicted structures, the identified zebrafish PGRP homologs resemble short and long PGRP members in arthropod and mammals. The identified PGRP genes in T. nigroviridis and TakiFugu rubripes resemble the long PGRPs, and the short PGRP genes have not been found in T. nigroviridis and TakiFugu rubripes databases. Computer modelling of these molecules revealed the presence of three alpha-helices and five or six beta-strands in all fish PGRPs reported in the present study. The long PGRP in teleost fish have multiple alternatively spliced forms, and some of the identified spliced variants, e.g., tnPGRP-L3 and tnPGRP-L4 (in: Tetraodon nigroviridis), exhibited no characters present in the PGRP homologs domain. The coding regions of zfPGRP6 (zf: zebrafish), zfPGRP2-A, zfPGRP2-B and zfPGRP-L contain five exons and four introns; however, the other PGRP-like genes including zfPGRPSC1a, zfPGRPSC2, tnPGRP-L1-, tnPGRP-L2 and frPGRP-L (fr: Takifugu rubripes) contain four exons and three introns. In zebrafish, long and short PGRP genes identified are located in different chromosomes, and an unknown locus containing another long PGRP-like gene has also been found in zebrafish, demonstrating that multiple PGRP loci may be present in fish. In zebrafish, the constitutive expressions of zfPGRP-L, zfPGRP-6 and zfPGRP-SC during ontogeny from unfertilized eggs to larvae, in different organs of adult, and the inductive expression following stimulation by Flavobacterium columnare, were detected by real-time PCR, but the levels and patterns varied for different PGRP genes, implying that different short and long PGRPs may play different roles in innate immune response. (c) 2007 Elsevier Ltd. All rights reserved.

A taxonomic study of Oconnorella (Enchytraeidae, Oligochaeta) from Changbaishan Mountain, China

We describe three enchytraeid species, including two new species, from Mt. Changbaishan, Jilin Province, northeastern China. Oconnorella cheni sp. nov. is characterized by a simple spermatheca; absence of the oesophageal appendages and seminal vesicle; and nephridia from 6/7, with a distinct funnel and the efferent duct arising from the anterior part of the postseptale. Oconnorella globula sp. nov. is distinguished by a spermatheca with two diverticula; lack of oesophageal appendages and seminal vesicle; and nephridia from 6/7, with distinct funnel and the efferent duct arising from the mid-ventral or posteroventral part of the postseptale. We redescribe Oconnorella changbaishanensis (Xie et al., 2000) from type and live specimens, and amend some characters that cannot be investigated clearly from mounted specimens. We revise the generic diagnosis of Oconnorella.

Garra rotundinasus, a new species of cyprinid fish (Pisces : Teleostei) from the upper Irrawaddy River basin, China

Garra rotundinasus, a new cyprinid species from the upper Irrawaddy River basin in Yunnan, China, is herein described. It shares with G. gravelyi the presence of a snout having a poorly developed proboscis represented by a truncate area in front of the nostrils, a character distinguishing both from all other Southeast Asian and Chinese congeners. The two species are distinct in coloration, morphometric and meristic characters. The sympatrically occurring G. tengchongensis is very similar to G. rotundinasus in possessing 36-37 perforated lateral line scales, 5 or 6 scales between the anus and anal-fin origin, and an anterior position of the anus (anus to anal distance 32.1-51.8% of pelvic to anal distance). Garra rotundinasus can be differentiated from G. tengehongensis in having a more slender caudal peduncle, a larger disc and no dark central band on the dorsal fin.

Reevaluation of the phylogenetic relationship between Mobilid and Sessilid peritrichs (Ciliophora, Oligohymenophorea) based on small subunit rRNA genes sequences

Based on morphological characters, peritrich ciliates (Class Olygohymenophorea, Subclass Peritrichia) have been subdivided into the Orders Sessilida and Mobilida. Molecular phylogenetic studies on peritrichs have been restricted to members of the Order Sessilida. In order to shed more light into the evolutionary relationships within peritrichs, the complete small subunit rRNA (SSU rRNA) sequences of four mobilid species, Trichodina nobilis, Trichodina heterodentata, Trichodina reticulata, and Trichodinella myakkae were used to construct phylogenetic trees using maximum parsimony, neighbor joining, and Bayesian analyses. Whatever phylogenetic method used, the peritrichs did not constitute a monophyletic group: mobilid and sessilid species did not cluster together. Similarity in morphology but difference in molecular data led us to suggest that the oral structures of peritrichs are the result of evolutionary convergence. In addition, Trichodina reticulata, a Trichodina species with granules in the center of the adhesive disc, branched separately from its congeners, Trichodina nobilis and Trichodina heterodentata, trichodinids without such granules. This indicates that granules in the adhesive disc might be a phylogenetic character of high importance within the Family Trichodinidae.

Revised diagnosis of the genus Bangana Hamilton, 1822 (Pisces : Cyprinidae), with taxonomic and nomenclatural notes on the Chinese species

Most morphological characters diagnostic of the 13 Chinese species of the cyprinid genus Sinilabeo Rendahl, 1932, are identical to those of the genus Bangana Hamilton, 1822. Consequently, these 13 species are transferred to Bangana. A revised diagnosis is provided for the now-expanded genus Bangana, and a dichotomous key and taxonomic and nomenclatural notes are included for the following valid Chinese species: B. decora, B. dero, B. devdevi, B. discognathoides, B. lemassoni, B. lippa, B. rendahli, B. tonkinensis, B. tungting, B. wui, B. xanthogenys, B. yunnanensis, and B. zhui. Literature reports, by Chinese authors, of Sinilabeo dero from the upper Irrawady River basin, in Yunnan, are based on misidentifcations of B. devdevi. Sinilabeo cirrhinoides Wu and Lin in Wu, Lin, Chen, Chen and He, 1977, and S. laticeps Wu and Lin in Wu, Lin, Chen, Chen and He, 1977, are junior subjective synonyms of B. devdevi and B. lippa, respectively. Sinilabeo yunnanensis Wu, Lin, Chen, Chen and He, 1977, is an available name, and a lectotype is designated for the species. Bangana zhui ( Zheng and Chen, 1983) is a valid species distinct from B. yunnanensis.

Redescription of Myxobolus pyramidis chen, 1958 (Myxosporea : Bivalvulida)

Redescription of Myxobolus pyramidis Chen, 1958, from gill lamellae of allogynogenetic gibel carp, Carassius auratus gibelio (Bloch), is presented in this paper to complete Chen's description. The diagnostic characters of the myxosporidia are: ovoid round, greyish-white polysporous plasmodia, averaging (159 +/- 21)x(72 +/- 6.5) mu m in size; spore pyriform in front view with smooth surface and symmetrical valves, convex-shaped in sutural view with straight and thick sutural line, averaging (10.5 +/- 1.1)x(10.3 +/- 0.9)x(6.1 +/- 0.2) mu m in size; two equal pyriform polar capsules averaging (5.5 +/- 0.7)x(3.5 +/- 0.2) mu m in size with distinct intercapsular process and polar filament wounded in five to six coils. The histological effects of the pathogen were observed by light microscopy, and the parasite-host relationship was discussed.

Acrossocheilus malacopterus, a new non-barred species of cyprinid from South China

Acrossocheilus malacopterus n. sp. is described from the Yuan Jiang (Red River) and Zhu Jiang basins in I South China. It is separated from other congeners by a combination of the following characters: an inconspicuously stripped body, 16 circumpeduncular scales, 47-49 lateral line scales, depth of caudal peduncle 7.8-9.3% SL, snout length 37.9-41.7% SL, dorsal-fin length 18.7-21.0% SL and last simple dorsal-fin ray slender with 8-10 fine serrations along posterior margin.

Phylogenetic relationships among Trichodinidae (Ciliophora : Peritricha) derived from the characteristic values of denticles

The phylogenetic relationships among trichodinids remain obscure. As an important diagnostic marker, the morphology of the denticles in the adhesive disc as well as the adoral spiral has been widely used in generic discrimination and species identification of trichodinids. We studied the characters of denticles of the ten genera of Trichodinidae and the sole genus Urceolaria of Urceolariidae by using a quantitative method. The characteristic values were used to generate Manhattan distance, on which the dendrogram was based to construct with the Unweighted Paired Group Method using the Arithmetic mean (UPGMA). The investigations show that all the genera of the family Trichodinidae were clearly separate from the outgroup Urceolaria, and within the Trichodinidae: (i) Dipartiella grouped with Trichodinella and Tripartiella and lay in the closest position to the outgroup with a low dissimilarity, suggesting Dipartiella might be the most primitive genus in the family; (ii) Hemitrichodina clustered in a single clad and lay in the farthest position to the outgroup with the highest dissimilarity, indicating that it might be the most advanced genus; and (iii) the other 6 genera, Trichodina, Paratrichodina, Semitrichodina, Vauchomia, Pallitrichodina and Trichodoxa clustered in a big clad with very low dissimilarity, showing that they are closely related to each other. We discuss the evolutionary trend of the denticle and conclude that the denticles of the adhesive disc should be an apomorphic feature of the trichodinids and their changes could reflect the evolutionary tendencies of these ciliates.

Garra bispinosa, a new species of cyprinid fish (Teleostei : Cypriniformes) from Yunnan, Southwest China

Garra bispinosa, a new species of cyprinid fish from the Irrawaddy River basin in Yunnan, Southwest China is described here. It has a conspicuous, quadrate and forwards pointed proboscis reflected downwards against the snout and anteriorly bilobed with one large, uniscupid and acanthoid tubercle on the distal end of each lobe, a character unique among all other congeners in Southeast Asia and China, and further differs from them in having the following combination of characters: snout with a deep groove across its tip to form a transverse lobe, 16 circumpeduncular scales, 34-35 lateral line scales, a smaller mental disc (length 38.1-43.8 % of head length), a forwards situated anus (anus to anal distance 25.9-30.6 % of pelvic to anal distance), and a slightly pointed snout.