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In order to explore the distribution characteristics of heavy metal contamination of farmland soil surrounding Luancheng town, Shijiazhuang City, Henan province, the concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn in the surface soil and crops were determined and assessed. The principal components were also analyzed for source apportionment. The heavy metal concentrations in crops were further detected, and the non-carcinogenic health risks in the study area were evaluated using the probabilistic risk assessment method, as to provide a theoretical basis for the treatment, prevention, and control of heavy metal pollution in farmland soil in Luancheng. According to the results

The Dianchi golden-line barbel, Sinocyclocheilus grahami (Regan, 1904), is one of the "Four Famous Fishes" of Yunnan Province, China. Given its economic value, this species has been artificially bred successfully since 2007, with a nationally selected breed (" S. grahami, Bayou No. 1") certified in 2018. For the future utilization of this species, its growth rate, disease resistance, and wild adaptability need to be improved, which could be achieved with the help of molecular marker-assisted selection (MAS). In the current study, we constructed the first chromosome-level genome of S. grahami, assembled 48 pseudo-chromosomes, and obtained a genome assembly of 1.49 Gb. We also performed QTL-seq analysis of S. grahami using the highest and lowest bulks (i.e., largest and smallest size) in both a sibling and random population. We screened two quantitative trait loci (QTLs) (Chr3, 14.9-39.1 Mb and Chr17, 4.1-27.4 Mb) as the major growth-related locations. Several candidate genes (e.g., map2k5, stat1, phf21a, sox6, and smad6) were also identified, with functions related to growth, such as cell differentiation, neuronal development, skeletal muscle development, chondrogenesis, and immunity. These results built a solid foundation for in-depth MAS studies on the growth traits of S. grahami.

The restocking of the endangered Kanglang white minnow (Anabarilius grahami) in Lake Fuxian, China, has been conducted for 13 years. However, few studies have reported on the effectiveness of the captive breeding and release of this species. Here, we investigated variations in morphology, including body shape and skeletal deformities, and genetic features among hatchery-born and recaptured A. grahami from Lake Fuxian. Results showed that current hatchery-reared fish displayed a stubbier body shape than their wild conspecifics from the 1980s. Furthermore, high skeletal deformity ratios were found in two aquafarms (Luchong, 50%; Haikou, 45.2%), and the release of malformed fish elevated the skeletal deformity rate of wild stocks found near the Lake Fuxian release sites (west coast, 19.0%; east coast, 12.5%). Based on variations in the cytochrome b (cyt b) gene, existing A. grahami populations showed relatively high haplotype diversity and low nucleotide diversity. Hatchery populations exhibited reduced genetic variations based on microsatellite markers and reintroduction led to markedly lower genetic diversity around the west coast release sites of Lake Fuxian. Analysis of molecular variance (AMOVA) of cyt b and microsatellite analysis showed that the greatest genetic variations were found within populations, and genetic distance and Bayesian clustering analysis showed that the 14 populations clustered into one group. Based on morphological and genetic tests, we discuss corresponding recommendations, including release size, feed formulations, breeding strategies, and release tags, to minimize potential risks and improve hatchery practices for better restocking of this species.

The origin and colonization history of Kurixalus, a genus of small arboreal tree frogs breeding exclusively in shallow swamps, is under disputed. On the basis of comprehensive sampling program, the evolutionary history of Kurixalus is investigated based on 3 mitochondrial genes. Our results indicate that the genus Kurixalus originated in the Asian mainland and subsequently arrived at its current distribution in Borneo, Taiwan, Ryukyu, and Hainan islands by a series of dispersal events. Moreover, the colonization of Taiwan from mainland Asia has occurred 2 times. The initial colonization of Taiwan occurred at 3.46–8.68 Mya (95% highest posterior density), which rejects the hypothesis that Kurixalus probably originated from Taiwan during the early Oligocene and favors the model of Neogene-origin rather than the model of Quaternary-origin for Taiwanese Kurixalus. Kurixalus eiffingeri has dispersed from Taiwan to the Ryukyus once or 2 times pending more data. Both transoceanic dispersal and landbridge dispersal have played a role in the colonization process; the former resulted in the colonization of Taiwan and the Ryukyus and the latter led to the colonization of Borneo and Hainan.

Pot and field trials were conducted to investigate Cd uptake and phytoremediation efficiency of two

The taxonomic history of Chinese semisulcospirids is confusing. Originally placed in Melania by late 19th century authors, species have subsequently been dispersed into four different genera, i.e. Semisulcospira Böttger, 1886, Senckenbergia Yen, 1939, Hua Chen, 1943, and Namrutua Abbott, 1948; however, these placements were not conducted within a phylogenetic context. Based on examination of newly collected samples from the Chinese provinces Yunnan, Guizhou, Jiangxi, Anhui and Chongqing, we revised the systematics of the Chinese Semisulcospiridae based on comparative anatomy and mitochondrial phylogenetics. We have examined shell, radular, and reproductive features and employed Bayesian and Maximum Likelihood analyses of partial sequences of mitochondrial genes cytochrome c oxidase I and 16S rRNA to resolve the phylogenetic relationships. The results revealed that Semisulcospiridae is represented by three genera in China, which differ in their reproductive strategy, including Semisulcospira, which is viviparous and contains three species (S. ningpoensis, S. gredleri, and S. pleuroceroides); Hua, which is oviparous and contains 11 species; and Koreoleptoxis, which is oviparous and contains one species. Namrutua and Senckenbergia are identified here as junior subjective synonyms of Semisulcospira. All taxa recognised herein formed well differentiated clades in three mitochondrial phylogeny, and could be reliably distinguished by differences in shell shape and sculpture. Four new species of Hua are described from Yunnan Province. This is the first comprehensive study to review the taxonomy of semisulcospirids in China. These species are sensitive to water quality, and economic development and infrastructure expansion have negatively impacted many populations. We fear that some species may have disappeared before being investigated or described scientifically. Therefore, this research will hopefully inspire further studies and surveys to foster our understanding of freshwater gastropod diversity in China. http://zoobank.org/urn:lsid:zoobank.org:pub:EF2147FD-9EEE-4922-90DC-76651EF9E6AE

The systematics of Semisulcospiridae in China is revised here based on morphological characters and mitochondrial phylogenetics. Phylogenetic relationships within the Chinese semisulcospirids were assessed via DNA sequences from mitochondrial analysis (cytochrome c oxidase I and 16S rRNA). This research contains most morphospecies of semisulcospirids previously recorded in China. Based on these results, the family of Chinese Semisulcospiridae is represented by three genera: i.e., viviparous Semisulcospira B?ttger, 1886, oviparous Hua Chen, 1943 and Koreoleptoxis Burch and Jung, 1988. These genera can be distinguished from each other by reproductive anatomy, reproductive mode, and radula features. Species of Hua are mainly distributed in southwest China and Guangxi, whereas Koreoleptoxis and Semisulcospira are mainly distributed in south and northeast China.

Intermuscular bones (IBs) are widely present in morphologically generalized teleost fishes and are commonly found in the Cyprinidae. Intermuscular bones are small, hard spicules of bone that are formed by ossification in the myosepta between neighboring myomeres. Why fish have IBs, and whether there is any evolutionary pattern to their occurrence, has been poorly understood. However, the presence of IBs does substantially affect the meat quality and commercial values of many cyprinid fishes in aquaculture. In this study, we sampled 592 individuals of cyprinid fishes to systematically investigate the evolution of IBs from a phylogenetic point of view. We found that the total number of IBs in the Cyprinidae ranged from 73 to 169, and we clarified that only two categories of IBs (epineural and epipleural) were present in all examined cyprinids. Most of the IBs were distributed in the posterior region of the fish, which might be an optimal target for selecting fewer IB strains in aquaculture. There was a positive correlation between IBs and the number of vertebrae, thus making it possible to predict the approximate number of IBs by counting the number of vertebrae. Although the IBs displayed some correlation with phylogenetic relationships in some lineages and to ecological factors such as diet (especially carnivore), in an overall view the variations of IBs in cyprinids were extremely diverse. The number and patterns of IBs in these fishes may reflect their phylogenetic history, but have been shaped by multiple environment factors. In this study, we also confirmed that X‐ray photography remains an optimal and reliable method for the study of IBs.

As one economically important fish in the southeastern Himalayas, the giant devil catfish (Bagarius yarrelli) has been known for its extraordinarily large body size. It can grow up to 2 m, whereas the non-Bagarius sisorids only reach 10–30 cm. Another outstanding characteristic of Bagarius species is the salmonids-like reddish flesh color. Both body size and flesh color are interesting questions in science and also valuable features in aquaculture that worth of deep investigations. Bagarius species therefore are ideal materials for studying body size evolution and color depositions in fish muscles, and also potential organisms for extensive utilization in Asian freshwater aquaculture. In a combination of Illumina and PacBio sequencing technologies, we de novo assembled a 571-Mb genome for the giant devil catfish from a total of 153.4-Gb clean reads. The scaffold and contig N50 values are 3.1 and 1.6 Mb, respectively. This genome assembly was evaluated with 93.4% of Benchmarking Universal Single-Copy Orthologs completeness, 98% of transcripts coverage, and highly homologous with a chromosome-level-based genome of channel catfish (Ictalurus punctatus). We detected that 35.26% of the genome assembly is composed of repetitive elements. Employing homology, de novo, and transcriptome-based annotations, we annotated a total of 19,027 protein-coding genes for further use. In summary, we generated the first high-quality genome assembly of the giant devil catfish, which provides an important genomic resource for its future studies such as the body size and flesh color issues, and also for facilitating the conservation and utilization of this valuable catfish.

A revision of Chinese freshwater Sulcospira gastropods is presented here based on analysis of the morphological characteristics and partial sequences of the mitochondrial genes 16S rRNA and cytochr...

Intermuscular bones (IBs) are slender linear bones embedded in muscle, which ossify from tendons through a process of intramembranous ossification, and only exist in basal teleosts. IBs are essential for fish swimming, but they present a choking risk during human consumption, especially in children, which can lead to commercial risks that have a negative impact on the aquaculture of these fish. In this review, we discuss the morphogenesis and functions of IBs, including their underlying molecular mechanisms, as well as the advantages and disadvantages of different methods for IB studies and techniques for breeding and generating IB-free fish lines. This review reveals that the many key genes involved in tendon development, osteoblast differentiation, and bone formation, e.g.,肌间刺（intermuscular bones，IBs）是仅存在于低等硬骨鱼类肌肉中的细长的线形骨骼， 由肌腱组织通过膜内骨化形成。虽然肌间刺在鱼类游泳过程中发挥重要作用，但是在人类食用过程中会导致潜在的卡喉咙的风险，尤其不方便儿童的食用，这也进一步影响了水产业的经济效益。该文综合阐述了肌间刺的形态发生和功能、相关发育分子机制、肌间刺不同研究方法，以及用于培育无肌间刺鱼类品系的相关技术。该综述揭示了许多与肌间刺发育相关的关键基因，它们主要参与肌腱发育、成骨细胞分化和骨形成发育，如scxa、msxC、sost、twist、bmps和osterix基因。因此，该文对如何利用基因组编辑和人工选育等办法培育无肌间刺的鱼类品系提供了有价值的信息。.

The loach tribe Yunnanilini from China is reviewed here using morphological characters and complete mitochondrial genomes of select species. Molecular data suggest that the tribe Yunnanilini is not monophyletic and can be divided into three clades. Species of the Yunnanilus nigromaculatus group form an independent genus and are placed in Eonemachilus. In the phylogenetic tree, Y. jinxiensis clusters with Paranemachilus genilepis, and Y. pulcherrimus clusters with Micronemacheilus cruciatus, indicating that Y. jinxiensis and Y. pulcherrimus belong to Paranemachilus and Micronemacheilus, respectively. Based on morphological data, Y. bailianensis and Y. longibarbatus are placed in Heminoemacheilus, while Y. jinxiensis and Y. pulcherrimus, are placed in Paranemachilus and Micronemacheilus, respectively. Yunnanilus niulanensis and Y. qujinensis are treated as junior synonyms of Eonemachilus caohaiensis. Eonemachilus, Micronemacheilus, and Yunnanilus are show short separation between anterior and posterior nostrils. The genera can be distinguished from each other by mouth structure, lateral line and cephalic lateral-line canals, and papillae on median part of both lips. The anterior and posterior nostrils of Heminoemacheilus and Paranemachilus are closely set. Paranemachilus and Micronemacheilus are distinguished by cheeks covered with scales and lips with papillae, respectively. Our phylogenetic tree and morphological characters support Traccatichthys as a valid genus, which can be distinguished from Micronemacheilus by anterior and posterior nostrils closely set (vs. clearly separated). Four species are placed in Traccatichthys.

The Dianchi golden-line barbel, Sinocyclocheilus grahami (Regan, 1904), is one of the “Four Famous Fishes” of Yunnan Province, China. Given its economic value, this species has been artificially bred successfully since 2007, with a nationally selected breed (“S. grahami, Bayou No. 1”) certified in 2018. For the future utilization of this species, its growth rate, disease resistance, and wild adaptability need to be improved, which could be achieved with the help of molecular marker-assisted selection (MAS). In the current study, we constructed the first chromosome-level genome of S. grahami, assembled 48 pseudo-chromosomes, and obtained a genome assembly of 1.49 Gb. We also performed QTL-seq analysis of S. grahami using the highest and lowest bulks (i.e., largest and smallest size) in both a sibling and random population. We screened two quantitative trait loci (QTLs) (Chr3, 14.9–39.1 Mb and Chr17, 4.1–27.4 Mb) as the major growth-related locations. Several candidate genes (e.g., map2k5, stat1, phf21a, sox6, and smad6) were also identified, with functions related to growth, such as cell differentiation, neuronal development, skeletal muscle development, chondrogenesis, and immunity. These results built a solid foundation for in-depth MAS studies on the growth traits of S. grahami.

In order to fully explore the spatial distribution of soil heavy metal contaminants in farmland soil surrounding a typical industrial area in Dakuai town, Xinxiang city, Henan Province, the concentrations of As, Cd, Cr, Cu, Ni, Pb, and Zn in the surface soil and within the soil profile were determined and assessed. The principal components were also analyzed for source apportionment to provide a theoretical basis for the control and prevention of heavy metal pollution. According to the results, the soils in the study area are severely contaminated by Cd and Cu and moderated contaminated by As due to the battery manufacture and Cu (e.g., pipe and wire) processing. The concentrations of Cd, Cu, and As in soils were (2.56±1.23) mg·kg

A pot experiment was conducted to investigate the growth response and Cd accumulation characteristics among different populations of

Analyses of morphological and molecular data from specimens ofHomatulafrom China reveal an undescribed species which lives in the Nanpanjiang River, the upper Pearl River, and indicate thatH. acuticephalais a synonym ofH. anguillioides. The new species is collected from Yunnan, China. It differs from its congeners by having a naked and robust body, developed adipose crests (depth of caudal peduncle in the percent of its length:70.53–78.48), a median notch on the lower jaw, a median gap on the lower lip, short barbels with maxillary barbels extend posteriorly reaching the anterior edge of eyes, vertebrae 37–38, completed lateral line. New species differs from other species ofHomatulaby 0.05–0.11 in K2P distance of mitochondrial cytochrome c oxidase subunit 1. Phylogenetic analysis based on mitochondrial cytochrome c oxidase subunit 1 indicate that new species formed a sister group to co-drainage species. Based on both morphological and molecular results, we confirm that the validation of this undescribed species namedH. robustasp. nov. and describe it.

The restocking of the endangered Kanglang white minnow (Anabarilius grahami) in Lake Fuxian, China, has been conducted for 13 years. However, few studies have reported on the effectiveness of the captive breeding and release of this species. Here, we investigated variations in morphology, including body shape and skeletal deformities, and genetic features among hatchery-born and recaptured A. grahami from Lake Fuxian. Results showed that current hatchery-reared fish displayed a stubbier body shape than their wild conspecifics from the 1980s. Furthermore, high skeletal deformity ratios were found in two aquafarms (Luchong, 50%; Haikou, 45.2%), and the release of malformed fish elevated the skeletal deformity rate of wild stocks found near the Lake Fuxian release sites (west coast, 19.0%; east coast, 12.5%). Based on variations in the cytochrome b (cyt b) gene, existing A. grahami populations showed relatively high haplotype diversity and low nucleotide diversity. Hatchery populations exhibited reduced genetic variations based on microsatellite markers and reintroduction led to markedly lower genetic diversity around the west coast release sites of Lake Fuxian. Analysis of molecular variance (AMOVA) of cyt b and microsatellite analysis showed that the greatest genetic variations were found within populations, and genetic distance and Bayesian clustering analysis showed that the 14 populations clustered into one group. Based on morphological and genetic tests, we discuss corresponding recommendations, including release size, feed formulations, breeding strategies, and release tags, to minimize potential risks and improve hatchery practices for better restocking of this species.

A hydroponic experiment was conducted to explore the differences in growth status and Cd accumulation characteristics of two

A blind fish of Sinocyclocheilus (Cypriniformes: Cyprinidae)was caught in open water in the Three Gorges (Sanxia)reservoir, at a depth of 20 m in the mainstream of YangtzeRiver in Zigui County, Hubei Province, China. This fish can beeasily distinguished from all other congeners by externalmorphological characteristics, and is estimated to havediverged from its sister group about 0.55 million years ago(Ma). The geologically well separated locality of this specieshas expanded the distribution of Sinocyclocheilus cavefishfrom around N25° (latitude) to above N30° . Herein, wedescribe this new species as Sinocyclocheilus sanxiaensis sp.nov ., and discuss the possible reasons why the speciesappears, surprisingly, in the Three Gorges reservoir.

Simple Summary Kanglang white minnow (KWM, Anabarilius grahami), is a typical “3E” (Endangered, Endemic and Economic) fish species in Yunnan-Guizhou Plateau. As one of the traditional “Four Famous Fishes” in Yunnan province, it has become the major local aquaculture species with increasing demand after the success of artificial breeding. However, this economically important fish is highly susceptible to the infection of a parasite ciliate, Ichthyophthirius multifiliis (Ich), during the practical procedure of artificial breeding. To examine the host immune responses to Ich, we divided the experimental fishes into three groups (including control, early-infected stage, and late-infected stage) for transcriptome sequencing to analyze the differentially expressed genes (DEGs) and immune response mechanisms. Abstract The notorious parasite Ichthyophthirius multifiliis (Ich) has been recorded worldwide in fish species and causes white spot disease, posing major threats and resulting in severe losses to international fish production. Extensively effective strategies for treating Ich are not available yet, and genetic mechanisms of hosts in response to the parasite are still largely unknown. In this study, we selected Kanglang white minnow (KWM, Anabarilius grahami) to examine its liver transcriptional changes after Ich infection, as white spot disease is one bottleneck problem in exploring this economically important species. We divided the experimental fishes into three groups (control, early-infected, and late-infected) to examine differentially expressed genes (DEGs). A total of 831 DEGs were identified and classified into 128 significantly enriched GO (Gene Ontology) terms and 71 significantly enriched KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways. Most of these terms or pathways were functionally enriched in immunity, inflammatory response, and apoptosis, such as nucleotide-binding oligomerization domain-like (NOD-like) receptor signaling, tumor necrosis factor (TNF) signaling, interleukin-17 (IL-17) signaling, and apoptosis pathways. We also identified 178 putative antimicrobial peptides (AMPs) and AMP precursors based on our previously reported genome assembly of KWM, and revealed that the expressional patterns varied according to different types. In summary, our work reported the first comprehensive transcriptional changes in KWM in response to the exogenous infection of Ich, which would lay a solid foundation for in-depth studies on disease defense or resistant strains selection in this valuable fish.

A new species of the genus Kurixalus (Anura: Rhacophoridae) is described from western Yunnan, China. Genetically the new species, Kurixalusyangi sp. n., is closer to Kurixalusnaso than to other known congeners. Morphologically the new species is distinguished from all other known congeners by a combination of the following characters: smaller ratios of head, snout, limbs, IND, and UEW to body size; male body size larger than 30 mm; curved canthus rostralis; weak nuptial pad; brown dorsal color; absence of large dark spots on surface of upper-middle abdomen; presence of vomerine teeth; gold brown iris; single internal vocal sac; serrated dermal fringes along outer edge of limbs; granular throat and chest; rudimentary web between fingers; and presence of supernumerary tubercles and outer metacarpal tubercle.

Hybridization has been successfully achieved using Dianchi golden-line barbel (Sinocyclocheilus grahami) and barbless carp (Cyprinus pellegrini) as parental species. However, little is known about the crossbreeding feasibility of S. grahami with domesticated common carp (Cyprinus carpio) and crucian carp (Carassius auratus). In this study, we constructed reciprocal hybrid lineages using parents of S. grahami (2n = 96) with C. carpio rubrofuscus (2n = 100) and C. auratus (2n = 100). We found that hybrids of S. grahami ♀ × C. carpio rubrofuscus ♂ survived, but with fertilization (41.6%) and hatching rates (68.2%) lower than the reciprocal hybrids (fertilization rate 92.7% and hatching rate 89.5%). Hybrids of S. grahami ♀ × C. auratus ♂ exhibited failed hatching and low fertilization (6.3%) compared with the reciprocal hybrids, which showed high fertilization (88.5%) and hatching rates (88.6%). Growth results show that hybrids of S. grahami ♀ × C. carpio ♂ (total length: 12.2 cm, body weight: 25.4 g), S. grahami ♂ × C. carpio ♀ (total length: 12.1 cm, body weight: 25.0 g), and S. grahami ♂ × C. auratus ♀ (total length: 10.1 cm, body weight: 16.7 g) achieved medium growth rates compared to their parents. Based on this study, the three hybrid lineages show fast growth performance compared to S. grahami and exhibit high commercial and aquacultural application potential.

Phytoextraction is a promising technology that uses hyperaccumulating plants to remove arsenic (As) and heavy metals (HMs) from soil. A field experiment has been conducted to evaluate impacts of water-soluble chitosan (WC) upon the growth and phytoextraction of As, Cd and Pb by the hyperaccumulator Pteris vittata L. and the economic plant castor bean ( Ricinus communis L.) in a co-planting system in polymetallic contamination of agricultural soil. The co-planting system was comprised of As-hyperaccumulator and Pb high-accumulating P. vittata and Cd high-accumulating R. communis . The results showed that yields of P. vittata were significantly increased by co-planting. The results revealed that the uptake of As and Pb by P. vittata was also significantly increased by co-planting. When the WC was applied to the co-planting system, the uptake of Cd and Pb by P. vittata and R. communis was significantly increased, but the uptake of As was not increased obviously. This study suggested that the combination of co-planting system with WC is a more promising approach than single hyperaccumulator. This approach not only enhanced the removal rates of soil As and HMs, but also simultaneously offered agricultural practices with no edible products in the As and HMs contaminated farmland soils.

Few empirical studies have compared coalescent-based methods to distance-based methods for delimitation of less divergent species. In this study, we used two coalescent-based (BFD and BPP) and two distance-based barcoding (ABGD and jMOTU) methods to delimit closely related species in the Kurixalus odontotarsus species group. Phylogenetic analyses revealed that the K. odontotarsus species group comprises 11 distinct maternal clades with strong support values. Based on the genetic and morphological evidences, we consider that species diversity in the K. odontotarsus species group was underestimated and the 11 clades represent 11 species, of which six are unnamed. The coalescent-based delimitations decisively supported the scenario of 11-species corresponding to the 11 clades. However, the distance-based ABGD only obtained 3–6 candidate species, which is not consistent with morphological evidence. These results indicate that BFD and BPP are more conservative than ABGD to false negatives (lumping). Method of fixed threshold (jMOTU) may obtain a resolution similar to that inferred by BFD and BPP, but it severely relies on subjective choice of the threshold and lacks statistical support. We consider that coalescent-based BFD and BPP approaches outperform distance-based methods for delineation of less divergent species.

Persistent economic growth in Chinese southwestern Yunnan Province is setting aquatic ecosystems in its plateau lakes under enormous pressure. While several different systems have previously been used to study these lakes, no existing methodology adequately measures both the chemical and biotic parameters of these water bodies. Here, we present a novel Biotic Monitoring Yunnan Lakes (BMYL) index that provides a general assessment tool for ecological deterioration that is caused by organic pollution. Principal Component Analysis is used to analyze the occurrence of families of macroinvertebrates and chemical properties of the lakes. In brief, families of macroinvertebrates were given a score from 10 to 1 based on sensitivity to organic enrichment and eutrophication. Sampling at each lake yielded an Average Score per Lake (ASPL) which is calculated by dividing the total BMYL by the total number of scoring families. High ASPL values characterize a biologically intact lake containing relatively large numbers of high scoring taxa, while lower ASPL values denote a polluted lake that does not support many high scoring taxa. The results of the BMYL show a notably more accurate characterization of the long-term health of concerned aquatic ecosystems than studies that use abundance levels of species or a simple analysis of chemical parameter.

Objective. To evaluate the efficacy and safety of Tongning Gel (TNG) compared to placebo-controlled (PC) for knee osteoarthritis (KOA). Methods. A multicentre, randomized, double-blinded, parallel, placebo-controlled, clinical trial was performed in 576 patients (432 patients in the TNG group, 144 patients in the PC group), and 1 in the experimental group withdrew due to nonuse of drug. Patients were randomized to receive TNG or PC applied to knee skin at 3g per time, 2 times per day, which lasted for 3 weeks. The Western Ontario and McMaster Universities Arthritis Index (WOMAC) pain score was used to evaluate the primary efficacy of TNG and WOMAC stiffness and physical function and total scores were used to evaluate the secondary efficacy of TNG. All participants who received at least one dose of study drug were included in the safety analysis. This trial has been registered in Chinese Clinical Trial Registry (no. CTR20131276). Results. Primary efficiency outcome: there were significant differences in the decreased value of WOMAC pain score between two groups (P<0.05), and the decreased value of WOMAC pain score in the TNG group were better than those in the PC group (P<0.05). Secondary efficiency outcome: the WOMAC total score, WOMAC stiffness score, WOMAC physical function score, and the decrease of the above indexes of the two groups of patients after treatment were statistically significant (P<0.05), and the improvement of the above indexes in the TNG group was better than that of the PC group (P<0.05). Safety Evaluation. A total of 42 adverse events were reported by 29 patients: 25 adverse events reported by 16 patients (3.71%) in the experimental group and 17 adverse events were reported by 13 patients (9.03%) in the control group. And 8 adverse reactions were reported by 6 patients including 2 adverse reactions by 2 patients (0.46%) in the experimental group and 6 adverse reactions by 4 patients (2.78%) in the control group. Two cases of significant adverse events occurred in the experimental group. Both groups had one serious adverse event, respectively, which were not relevant to the intervention. Conclusion. These results of the trial demonstrate that TNG is superior to placebo in the treatment of patients with KOA, and TNG can improve other symptoms of KOA, such as stiffness and physical function. TNG is safe for the treatment of knee osteoarthritis as a whole.

We describe a new species of Zhangixalus from southern Yunnan of China, Vietnam, and Thailand based on morphological and molecular evidence. The new species had been confused with Zhangixalus smaragdinus (Blyth, 1852) in the past. Zhangixalus pachyproctus sp. nov. can be distinguished from Z. smaragdinus morphologically by the protruding vent in adult males, large thick grey reticular mottles below the white stripe on flank, more oblique snout in profile and wider head, longer snout, greater internarial distance, larger tympanum and longer hindlimb. The new species can be distinguished from other species of Zhangixalus with green dorsum by the following combination of characters: body size larger (SVL of adult males: 74.2–83.3 mm; SVL of adult female: 102.4 mm); dorsum smooth; narrow white stripes along edge of the lower jaw, body sides, outer side of limbs and above the vent; absence of brown bands on canthus rostralis, upper eyelid and supratympanic fold; webbing between fingers and toes complete except between the first two fingers; and internal single subgular vocal sac. Phylogeny based on comparison of 16S rRNA sequences suggests that the new species is the sister taxon to Z. smaragdinus and the two species differ by 7.63% in the uncorrected pairwise distance of 16S sequences.

In this study, pure‐breeding and crossbreeding of Sinocyclocheilus grahami were established to test heterosis and expression of growth‐related genes which were examined in the first‐generation (F1). Genetic distances results showed that Muyang river (M) and Sanbaimu reservoir (S) populations had smaller interspecific distances (genetic distance: 0.000) while larger distances were shown to other two populations (0.024 of Jiuzhai vauclusian spring [J] and 0.044 of Chenglong vauclusian spring [C]). Full‐sib families were created by using the Muyang river population as females: MM, MS, MJ and MC. Growth performance had significant differences between each group of F1 hybrids; the fastest growth group was shown in MJ while the slowest was in MS. For growth‐related genes, the fast growth group of MJ hybrids showed significantly higher levels of ghrelin expression in strong expression tissues of the intestine and liver. Among the four combinations, mRNA levels of insulin‐like growth factor I (IGF‐I) were the highest expression in MJ hybrids than others in all tested tissues. Myostatin b and somatostatin1 (SST1) were strongly expressed in the brain and MJ hybrids showed relatively lower expression level. In the muscle, slow growth group of MS hybrids showed extremely higher expression levels of these two genes, especially SST1 gene. In conclusion, associating growth with growth‐related genes expression indicated faster growth of crossbreeding in S. grahami and can make a contribution to commercial fish production in the future.

We record Raorchestes parvulus (Boulenger, 1893) for the first time from China based on six specimens collected from southern Yunnan. Phylogenetically these individuals were placed in a clade with R. parvulus from Southeast Asia and pairwise genetic distances in DNA sequences of the mitochondrial 16S rRNA gene ranged from 0 to 3.1% within the clade. Morphologically these specimens agree with R. parvulus in a series of characters including body size small, a large external vocal sac, vomerine teeth absent, snout slightly shorter than the diameter of the orbit, canthus rostralis indistinct, interorbital space broader than the upper eyelid, tympanum indistinct, fingers free, toes webbed at the base, an inner metatarsal tubercle, tibio-tarsal articulation reaches eye, small warts on the dorsal surfaces of body and limbs, throat smooth, belly granular, a dark triangular blotch between the eyes, a butterfly-like dark band on the back, a dark blotch on each side of lumbar region, and more or less distinct dark bars across limbs. Obvious intraspecific variation in the color pattern occurs within the population. It is possible that previous records of Raorchestes longchuanensis (Yang and Li, 1979) from southern Yunnan might be misidentification of R. parvulus.

A new cave-dwelling loach of the genus Triplophysa, T. xichouensis, is described from an outlet of a subterranean river in Xisa Town, Xichou County, Yunnan Province, China. It can be distinguished from its congeners by the following characters: dorsal-fin rays iii, 8; anal-fin rays ii, 6; pectoral-fin rays i, 9 or 10; pelvic-fin rays i, 5 or 6; branched caudal-fin rays 16(8+8); eyes highly degenerated to a very tiny black dot; dorsal-fin origin closer to snout tip than to caudal-fin base and anterior to vertical line of pelvic-fin origin; pectoral fin length about two-thirds the distance between pectoral-fin origin to pelvic-fin origin; caudal peduncle slender, its length about three times its depth; caudal fin emarginate; body smooth and scaleless; lateral line complete and straight; anterior chamber of air bladder wrapped in dumbbell-shaped bony capsule and the posterior one well developed, long, oval; intestine short, bending in zigzag shape behind stomach. A key for the cave-dwelling species of Triplophysa is provided. urn:lsid:zoobank.org:pub:9162FFB1-7911-47C3-AE50-6A00E9590327

Free Cu species in soils is a key issue to its bioavailability. However, predictive models for Cu speciation across a wide range of soils were still unavailable. In this study, Cu speciation in 34 contaminated soil samples were investigated via analytical technique and predictive models. The results showed that most of free Cu2+ was underestimated when using default log KCuFA and 65% active fulvic acid as inputs in models of WHAM VI and NICA-Donnan. The best prediction was found when using either adjusted active fulvic acid from 10% to 125% for WHAM VI or from 15% to 65% for NICA-Donnan model with the RMSE 0.96. In contrast, NICA-Donnan demonstrated a slightly stronger binding for Cu than WHAM VI due to extra 26% of samples was underestimated. This work presents a comprehensive database of Cu speciation and an effective attempt of free Cu2+ prediction in a wide range of Chinese soils.

Semisulcospira paludiformis, collected from Hainan Island, China, was described by Yen in 1939. While the type specimens of S. paludiformis were destroyed in World War II, the systematic classifica...

The taxonomy and phylogeny of the Chinese species of the Barbinae (Cypriniformes) has a confusing history. In this study, partial sequences of four mitochondrial genes (cyt b, COI, ND4 and 16S rRNA) from 75 Barbinae species and 38 outgroup species were used to investigate the taxonomy and phylogeny within the Barbinae in China. The monophyly of Neolissochilus, Sikukia and Tor are not supported. Neolissochilus benasi might represent a new genus, and Tor hemispinus and Tor qiaojiensis should be moved into Neolissochilus. Sikukia flavicaudata is not Sikukia species. Puntius paucimaculatus might be a synonym of Puntius semifasciolatus. Puntius semifasciolatus does not belong to Puntius. Onychostoma barbatum might consist of more than one species. Our molecular results corroborate that Acrossocheilus stenotaeniatus is a synonym of Acrossocheilus longipinnis. Finally, Barbonymus gonionotus from Menglun, Yunnan should be Poropuntius huangchuchieni.

Biological invasion is a pervasive negative force of global change, especially in its effects on sensitive freshwater ecosystems. Even protected areas are usually not immune. Ptychobarbus chungtienensis is a threatened freshwater fish now almost confined to Bita Lake, in the Shangri-La region of Yunnan province, China. Its existence is threatened by the introduction of non-native weatherfishes (Misgurnus anguillicaudatus and Paramisgurnus dabryanus) by an unusual method known as ‘prayer animal release’. Periodic surveys revealed the ratio of invasive weatherfishes to P. chungtienensis has been increasing since the former species was first recorded from the lake in August, 2009. Ptychobarbus chungtienensis shows low genetic diversity in the relict Lake Bita population. Weatherfishes, however, have highly successful survival strategies. The degree of dietary overlap between the species is alarming and perhaps critical if food is found to be a limiting factor.

A hydroponic experiment was conducted to study the interaction of Cd and Zn affecting the root morphology and accumulation of Cd and Zn in

A pot experiment was conducted to assess the phytoextraction potential for cadmium (Cd) of three types of sunflowers (edible, ornamental, and oil sunflowers) and the effects of the application of N, NP, and NPK fertilizers on Cd uptake of the three plants using Cd-contaminated soils collected from northern China. The results showed that fertilization could significantly increase the biomass, aboveground Cd concentrations and accumulation of ornamental and oil sunflowers, and the effect of NPK fertilizer was significantly better than those of other treatments. Cd concentrations of the aboveground parts of edible, ornamental, and oil sunflowers were 6.89, 8.92, 6.97 mg·kg