Balitora nantingensis (Teleostei : Balitoridae), a new hillstream loach from Salween drainage in Yunnan, Southwestern China

A new species of hillstream loach genus Balitora Gray is discovered from Nanting River drainage, which drains to Salween, Yongde County, Yunnan, China. Balitora nantingensis can be distinguished from B. lancangjiangensis by a triangle-like blotch in nape absent vs. present; area from halfway of pelvicfin ray to anus covered by scales vs. no scales in this area; the first pectoral ray thickened vs. not; posterior end of pelvic-fin surpasses middle point from posterior end of pelvic-fin base to anal-fin origin vs. the end from posterior extremity of pelvic-fins to anus; fewer lateral line scales, 59-64 vs. 66-70; slightly longer head, SL 4.2-5.1 (4.6) times of HL vs. 5.0-5.9 (5.4); slightly larger eyes, HL 4.0-6.4 (5. 1) times of eye diameter vs. 5.5-7.5 (6.3). Balitora nantingensis can be distinguished from B. burmanica by body rounded vs. slender in dorsal view; pointed snout vs. blunt in dorsal view; a deeper body, body depth 12.5-16.9% of SL vs. 10-12%; a deeper caudal-peduncle, length of caudal-peduncle 2.0-2.8 times of its depth vs. 3.0-4.2; a shorter caudal-peduncle, 14.3-15.6% of SL vs. 16-21%; the average ratio of body width at pelvic-fin origin and distance between inner pelvic-fin base 2.0 vs. 3.4; body width 1.1-1.5 times of body depth vs. 1.5-1.6; posterior end of pelvic-fin almost one or slight more than one eye diameter from anus vs. closet to anus; outside rays of pelvic-fin extend more posteriorly than inside rays and make posterior outline of fin acclivitous and straight vs. the outside ones almost equal to the inside ones and make the fin slightly convex.

Schistura cryptofasciata, a new loach (Cypriniformes : Balitoridae) from Salween drainage in Yunnan, Southwestern China

Schistura cryptofasciata is distinguished from its congeners by following combination characters: lateral line complete; 8(1)/(2) branched dorsal-fin rays; 9+8 branched caudal-fin rays; no sexual dimorphism; 8-11 indistinct bars, wider than interspaces; caudal-peduncle length 44.1-16.5% standard length, 68.1-90.4% dorsal head length; anterior portion of body scaleless; air bladder entirely embedded in bony capsule; lateral head length 104-122% dorsal head length; lower jaw with a notch at symphysis; lower lip completely interrupted; interorbital width 1.9-2.2 times eye diameter; pelvic-fin origin ventral to dorsal-fin origin; anus 1.0-1.5 eye diameters in front of anal-fin origin; the widest part of black basal caudal bar is the midlateral, and then the upper and lower ends.

A new species of sisorid catfish genus Glyptothorax (Teleostei: Sisoridae) from Salween drainage of Yunnan, China

A new species of the genus Glyptothorax, Glyptothorax obliquimaculatus sp. nov. is described from the Xiaohei River, a tributary of the Nanting River, Salween drainage, in southwestern Yunnan province, China. This new species can be distinguished from its congeners by the following combination of characteristics: unculiferous ridges of the thoracic adhesive apparatus extending anteriorly onto the gular region; body with irregular dark blotches scattered along lateral surface (blotches mostly oblique); skin smooth on head and body; dorsal spine smooth without serrations on its posterior margin; lips smooth; posterior margin of pectoral spine with 7-8 serrations; dorsal-fin base 11.0-13.2% SL; pectoral-fin length 15.6-19.6% SL; depth of caudal peduncle 8.6-9.8% SL; head width 19.1-24.0% SL; nasal barbel length 23.3-33.3% HL.

Fish-Specific Duplicated dmrt2b Contributes to a Divergent Function through Hedgehog Pathway and Maintains Left-Right Asymmetry Establishment Function

Gene duplication is thought to provide raw material for functional divergence and innovation. Fish-specific dmrt2b has been identified as a duplicated gene of the dmrt2a/terra in fish genomes, but its function has remained unclear. Here we reveal that Dmrt2b knockdown zebrafish embryos display a downward tail curvature and have U-shaped somites. Then, we demonstrate that Dmrt2b contributes to a divergent function in somitogenesis through Hedgehog pathway, because Dmrt2b knockdown reduces target gene expression of Hedgehog signaling, and also impairs slow muscle development and neural tube patterning through Hedgehog signaling. Moreover, the Dmrt2b morphants display defects in heart and visceral organ asymmetry, and, some lateral-plate mesoderm (LPM) markers expressed in left side are randomized. Together, these data indicate that fish-specific duplicated dmrt2b contributes to a divergent function in somitogenesis through Hedgehog pathway and maintains the common function for left-right asymmetry establishment.

Toll-like receptor 4 signaling pathway can be triggered by grass carp reovirus and Aeromonas hydrophila infection in rare minnow Gobiocypris rarus

Toll-like receptor 4 (TLR4) is critical for LPS recognition and cellular responses. It also recognizes some viral envelope proteins. Detection mostly results in the inflammation rather than specific antiviral responses. However, it's unclear in fish. In this report, a TLR4 gene (named as GrTLR4b) was cloned and characterized from rare minnow Gobiocypris rarus. The full length of GrTLR4b cDNA consists of 2766 nucleotides and encodes a polypeptide of 818 amino acids with an estimated molecular mass of 94,518 Da and a predicted isoelectric point of 8.41. The predicted amino acid sequence comprises a signal peptide, six leucine-rich repeat (LRR) motifs, one leucine-rich repeat C-terminal (LRRCT) motif, followed by a transmembrane segment of 23 amino acids, and a cytoplasmic region of 167 amino acids containing one Toll - interleukin 1 - receptor (TIR) motif. It's closely similar to the zebrafish (Danio rerio) TLR4b amino acid sequence with an identity of 77%. Quantitative RT-PCR analysis showed GrTLR4b mRNA was constitutive expression in gill, heart, intestine, kidney, liver, muscle and spleen tissues in healthy animals and up-regulated by viruses and bacteria. After being infected by grass carp reovirus or Aeromonas hydrophila, GrTLR4b expressions were up-regulated from 24 h post-injection and lasted until the fish became moribund (P < 0.05). These data implied that TLR4 signaling pathway could be activated by both viral and bacterial infection in rare minnow. (C) 2009 Elsevier Ltd. All rights reserved.

Population genetic structure and geographical differentiation of the Chinese catfish Hemibagrus macropterus (Siluriformes, Bagridae): Evidence for altered drainage patterns

National Natural Science Foundation of China (NSFC) [2007CB411600, 30530120]

Pseudobagrus fui Miao, a valid bagrid species from the Yangtze River drainage, South China (Teleostei: Bagridae)

Pseudobagrus fui Miao is a valid bagrid species that can be separated from all its congeners in having 27-33 anal-fin rays. Among the species of Pseudobagrus, it forms part of a group of approximately 20 species putatively characterized by having a smooth anterior margin of the pectoral-fin spine and short maxillary barbels not extending to the base of the pectoral-fin spine. Pseudobagrus fui, together with P. nitidus and P. vachelli, can be differentiated from all other fork-tailed species of this group by having no fewer than 20 branched anal-fin rays, the posterior end of the anal-fin base posterior to the vertical through the posterior end of the adipose-fin base, and anal-fin base longer than the adipose-fin base. It further differs from P. nitidus in having a gas bladder without beaded lateral and posterior margins, and from P. vachelli in having short maxillary barbels not extending to the base of the pectoral-fin spine. Pseudobagrus fui is currently known from the main stream of the upper Yangtze River and its tributaries, the Min River, Jialing River, Tuo River, Wu River and Chishui River in Sichuan Province, Guizhou Province, and the Chongqing City. The identity and nomenclature of Pseudobagrus nitidus is also discussed.

Grass carp reovirus activates RNAi pathway in rare minnow, Gobiocypris rarus

Dicer catalyzes the initiation step of RNA interference (RNAi) which is known to play a significant role in innate immune response to viral infection in many organisms. To study the RNAi-related pathway after virus infection in fish, we identified a partial cDNA sequence of dicer from rare minnow, Gobiocypris rants. Real-time quantitative RT-PCR (qRT-PCR) demonstrated the Dicer transcript level was the highest at zygote stage, decreased at prim-5 stage, and was stable from the protruding mouth to adult stage. Regular RT-PCR analysis showed that the Dicer gene expressed widely in the tested tissues, including brain, gill, heart, intestine, kidney, liver, muscle, ovary, spleen and testis. The expression of Dicer mRNA was significantly increased in the early period of Grass carp reovirus (GCRV) infection, and declined from 24 It post-injection (h p.i.) (P<0.05). The mRNA expression returned to control levels at 48 h p.i. (P>0.05). Under transmission electron microscope, virions were difficulty to find out in 12 h p.i., and virus inclusion bodies and few scattered viral particles were easily visualized from 24 h p.i. to moribund. These results implied GCRV triggered the RNAi pathway in the early stages of infection and perhaps virus inclusion bodies suppressed the antiviral functions of RNAi mechanism. (C) 2009 Published by Elsevier B.V.

Isolation and characterization of Argonaute 2: A key gene of the RNA interference pathway in the rare minnow, Gobiocypris rarus

Argonaute 2 gene plays a pivotal role in RNAi in many species. Herein is the first report of the cloning and characterization of Argonaute 2 gene in fish. The full-length cDNA of Gobiocypris rarus Argonaute 2 (GrAgo2) consisted of 3073 nucleotides encoding 869 amino acid residues with a calculated molecular weight of 98.499 kDa and an estimated isoelectric point of 9.18. Analysis of the deduced amino acid sequence showed the presence of two signature domains, PAZ and Piwi. RT-PCR analysis indicated that GrAgo2 mRNA expression could be detected in widespread tissues. After infection with grass carp reovirus, GrAgo2 expression was up-regulated from 12 h post-injection (p < 0.05) and returned to control levels at 48 h post-injection (p > 0.05). These data imply that GrAgo2 is involved in antiviral defense in rare minnow. (C) 2008 Published by Elsevier Ltd.

Metzia longinasus, a new cyprinid species (Teleostei: Cypriniformes) from the Pearl River drainage in Guangxi Province, South China

Metzia longinasus, a new cultrine cyprinid species from the Hongshui-He River of the Pearl River drainage in Guangxi Province, southern China, is described here. It can be distinguished from all other congeners by having a combination of the following characters: a superior mouth, 43 or 44 lateral-line scales, a long head (length 27.2-30.8% of standard length), a long snout (length 31.7-37.0% of head length), 10 or 11 branched anal-fin rays, and no longitudinal black band extending along each side of body from the extremity of the gill opening to caudal-fin base.

RNAi suppression of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) mediated differentially expressed genes involved in Toll-like receptor signaling pathway and caused increased susceptibility to Flavobacterium columnare

In Drosophila, Toll signaling cascade, which resembles the mammalian Toll-like receptor (TLR)/IL-1R signaling pathways and regulates the expression of anti-microbial peptide genes, mainly relies on peptidoglycan recognition proteins (PGRPs) for the detection of bacterial pathogens. To explore the effect of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) on Toll-like receptor signaling pathway, RNA interference (siRNA) and real time quantitative PCR (RQ-PCR) methods were used to identify differentially expressed genes regulated by zfPGRP6. The target genes included TLR2, TLR3, TLR5, TLR7, TLR8, IL1R, Sterile-alpha and Armadillo motif containing protein (SARM), myeloid differentiation factor 88 (MyD88) and nuclear factor (NF)-kappa B2 (p100/p52). The results of RQ-PCR showed that RNAi-mediated Suppression of zfPGRP6 significantly down-regulated the expression of TLR2, TLR5, IL1R, SARM, MyD88 and p100/p52. The expression of beta-defensin-1 was also down-regulated in those embryos silenced by zfPGRP6. In challenge experiments to determine the anti-bacterial response to Gram-negative bacteria, RNAi knock-down of zfPGRP6 markedly increased susceptibility to Flavobacterium columnare. (C) 2008 Elsevier B.V. All rights reserved.

DE-71-induced apoptosis involving intracellular calcium and the Bax-mitochondria-caspase protease pathway in human neuroblastoma cells In vitro

Polybrominated diphenyl ethers (PBDEs) are used extensively as flame-retardants and are ubiquitous in the environment and in wildlife and human tissue. Recent studies have shown that PBDEs induce neurotoxic effects in vivo and apoptosis in vitro. However, the signaling mechanisms responsible for these events are still unclear. In this study, we investigated the action of a commercial mixture of PBDEs (pentabrominated diphenyl ether, DE-71) on a human neuroblastoma cell line, SK-N-SH. A cell viability test showed a dose-dependent increase in lactate dehydrogenase leakage and 3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyl-tetrazolium bromide reduction. Cell apoptosis was observed through morphological examination, and DNA degradation in the cell cycle and cell apoptosis were demonstrated using flow cytometry and DNA laddering. The formation of reactive oxygen species was not observed, but DE-71 was found to significantly induce caspase-3, -8, and -9 activity, which suggests that apoptosis is not induced by oxidative stress but via a caspase-dependent pathway. We further investigated the intracellular calcium ([Ca2+](i)) levels using flow cytometry and observed an increase in the intracellular Ca2+ concentration with a time-dependent trend. We also found that the N-methyl d-aspartate (NMDA) receptor antagonist MK801 (3 mu M) significantly reduced DE-71-induced cell apoptosis. The results of a Western blotting test demonstrated that DE-71 treatment increases the level of Bax translocation to the mitochondria in a dose-dependent fashion and stimulates the release of cytochrome c (Cyt c) from the mitochondria into the cytoplasm. Overall, our results indicate that DE-71 induces the apoptosis of ([Ca2+](i)) in SK-N-SH cells via Bax insertion, Cyt c release in the mitochondria, and the caspase activation pathway.

Pseudobagrus brachyrhabdion, a new catfish (Teleostei : Bagridae) from the middle Yangtze River drainage, South China

Pseudobagrus brachyrhabdion sp. nov., from the Yuan Jiang and Xiang Jiang of the middle Yangtze River drainage in Hunan and Guizhou Provinces, South China, is described herein. It is distinguished from all other Pseudobagrus species with a truncate or slightly emarginated caudal fin by an unique combination of the following characters: supraoccipital plate and nuchal plate broadly interspaced and covered with skin; nasal barbels only at most reaching anterior margin of eye; maxillary barbels reaching slightly beyond posterior margin of eye; outer mandibular barbels extending to posterior margin of eye; dorsal fin with a somewhat convex distal margin, origin nearer to pectoral-fin insertion than to pelvic-fin insertion; dorsal-fin spine shorter than pectoral spine, with a somewhat serrated posterior margin; pectoral-fin spine with a smooth anterior margin; anal fin with 20-23 rays, base length 23.8-32.0% of standard length, posterior end of anal-fin base anterior to posterior end of adipose fin base; no longitudinal black band extending along flank; eyes large, diameter 16.3-23.7% of head length; and number of vertebrae 5 + 43-46.

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.

Use of microbial community to evaluate performance of a wetland system in treating Pb/Zn mine drainage

The performance of a wetland system in treating lead (Pb)/zinc (Zn) mine drainage was evaluated by using the polyurethane foam unit (PFU) microbial community (method), which has been adopted by China as a standardized procedure for monitoring water quality. The wetland system consisted of four cells with three dominant plants: Typha latifolia, Phragmites australis and Paspalum distichum. Physicochemical characteristics [pH, EC, content of total suspended solid (TSS) and metals (Pb, Zn, Cd, and Cu)] and PFU microbial community in water samples had been investigated from seven sampling sites. The results indicated that the concentrations of Pb, Zn, Cd, Cu, and TSS in the mine drainage were gradually reduced from the inlet to the outlet of the wetland system and 99%, 98%, 75%, 83%, and 68% of these metals and TSS respectively, had been reduced in concentration after the drainage passed through the wetland system. A total of 105 protozoan species were identified, the number of protozoa species and the diversity index (DI) gradually increased, while the heterotrophic index (HI) gradually decreased from the inlet to the outlet of the wetland system. The results indicated that DI, HI, and total number species of protozoa could be used as biological indicators indicating the improvement of water quality.