Evolutionary Conservation of Dazl Genomic Organization and its Continuous and Dynamic Distribution Throughout Germline Development in Gynogenetic Gibel Carp

To investigate germline development and germ cell specification, we identified a Dazl homolog (CagDazl) from gynogenetic gibel carp (Carassius auratus gibelio). Its cDNA sequence and BAC clone sequence analyses revealed the genomic organization conservation and conserved synteny of the Dazl family members and their neighborhood genes among vertebrates, especially in fish. Moreover, a polyclonal antibody specific to CagDazl was produced and used to examine its expression and distribution throughout germline development at protein level. Firstly, ovary-specific expression pattern of CagDazl was confirmed in adult tissues by RT-PCR and Western blot. In addition, in situ hybridization and immunofluorescence localization demonstrated its specific expression in germ cells, and both its transcript and protein were localized to germ plasm. Then, co-localization of CagDazl and mitochondrial cloud was found, confirming that CagDazl transcript and its protein are germ plasm component and move via METRO pathway during oogenesis. Furthermore, the CagDazl is abundant and continuous throughout germline development and germ cell specification including primordial germ cell (PGC) formation, oogonium differentiation, oocyte development, and embryogenesis, and the dynamic distribution occurs at different development stages. The data suggest that maternal CagDazl might play an important role in gibel carp PGC formation. Therefore, CagDazl is a useful and specific marker for tracing germ plasm and germ cell development in the gynogenetic gibel carp. In addition, in comparison with previous studies in sexual reproduction species, the continuous and dynamic distribution of CagDazl protein in the germ plasm throughout the life cycle seems to have significant implication in sex evolution of vertebrates. J. Exp. Zool. (Mol. Deu. Euol.) 312B:855-871, 2009. (C) 2009 Wiley-Liss, Inc.

Effects of damming on population sustainability of Chinese sturgeon, Acipenser sinensis: evaluation of optimal conservation measures

The numbers of spawning sites for Chinese sturgeon have been drastically reduced since the construction of the Gezhouba Dam across the Yangtze River. This dam has blocked migration of Chinese sturgeon to their historic spawning ground causing a significant decline of the Chinese sturgeon population. We conducted a VORTEX population viability analysis to estimate the sustainability of the population and to quantify the efficiency of current and alternative conservation procedures. The model predicted the observed decline of Chinese sturgeon, resulting from the effect of the Gezhouba Dam. These simulations demonstrated the potential interest of two conservation measures: increasing spawning area and reducing predation on sturgeon eggs. The simulations also demonstrated that the actual restocking program is not sufficient to sustain sturgeon population as the artificial reproduction program induce the loss of more wild mature adults that the recruitment expected by the artificial reproduction.

Population status, threats and conservation of the Yangtze finless porpoise

The Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis) is currently limited to the middle and lower reaches of the Yangtze River from Yichang to Shanghai, China, and the adjoining Poyang and Dongting Lakes. Its population size has decreased remarkably during the last several decades due to the heavy impact of human activities, including overfishing of prey species, water development projects that cause attendant habitat loss and degradation, water pollution, and accidental deaths caused by harmful fishing gear and collisions with motorized vessels. It was estimated that the number of remaining individuals was down to approximately 1800 in 2006, a number that is decreasing at a rate as high as 5% per year. Three conservation measures - in situ and ex situ conservation and captive breeding have been applied to the protection of this unique porpoise since the early 1990s. Seven natural and two "semi-natural" reserves have so far been established. Since 1996, a small group of finless porpoises has been successfully reared in a facility at the Institute of Hydrobiology of the Chinese Academy of Sciences; three babies were born in captivity on July 5, 2005, June 2, 2007 and July 5, 2008. These are the first freshwater cetaceans ever born in captivity in the world. Several groups of these porpoises caught in the main stream of the Yangtze River, or rescued, have been introduced into the Tian'e-Zhou Semi-natural Reserve since 1990. These efforts have proven that, not only can these animals survive in the area, they are also to reproduce naturally and successfully. More than 30 calves had been born in the reserve since then, with one to three born each year. Taking deaths and transfers into account, there were approximately 30 individuals living in the reserve as of the end of 2007. Among eight mature females captured in April 2008, five were confirmed pregnant. This effort represents the first successful attempt at off-site protection of a cetacean species in the world, and establishes a solid base for conservation of the Yangtze finless porpoise. A lesson must be drawn from the tragedy of Chinese River Dolphin (Lipotes vexillifer), which has already been declared likely extinct. Strong, effective and appropriate protective measures must be carried out quickly to prevent the Yangtze finless porpoise from becoming a second Chinese River Dolphin, and save the biodiversity of the Yangtze River as a whole.

Effects of constructed wetland system on the removal of dibutyl phthalate (DBP)

Phthalic acid esters (PAEs) have become widely diffused in the environment via the manufacturing process. Numerous experiments have shown that the bioaccumulation of PAEs occurred in the aquatic and terrestrial food chain; meanwhile, it was found that some of PAEs were considered as potential carcinogens, teratogens and mutagens. In this research, two vertical/reverse-vertical flow constructed wetland systems were set up to study its removal efficiency of dibutyl, phthalate (DBP) pollution. The results showed that the constructed wetland system could remove DBP effectively, and the removal rates reached nearly 100%. Substrate microorganism and enzymatic activities probably played key roles during DBP removal, and the removal of DBP probably mainly took place in the upper layer of chamber A in the constructed wetland systems. (C) 2007 Elsevier GmbH. All rights reserved.

Abundance and conservation status of the Yangtze finless porpoise in the Yangtze River, China

middle and lower reaches of the Yangtze River, China. It is the only freshwater population of porpoises in the world and is currently listed as Endangered by IUCN. In November and December 2006 we used two boats and line transect methods to survey the entire current range of the population, except for two lakes (Poyang and Dongting). Sighting results were similar for both boats, so we pooled all data and analyzed them using two line transect models and a strip transect model. All models produced similar estimates of abundance (1111, 1225 and 1000). We then added independent estimates of the number of porpoises from the two lakes for a total estimate of approximately 1800 porpoises. Our findings indicate that the population continues to decline and that its distribution is becoming more fragmented. Our current estimate in the main river is slightly less than half the estimate from surveys between 1984 and 1991 (which was probably an underestimate). We also found an apparent gap in the distribution of porpoises between Yueyang and Shishou (similar to 150 km), where sightings had previously been common. Continued threats to Yangtze finless porpoises include bycatch in unregulated and unselective fishing, habitat degradation through dredging, pollution and noise, vessel strikes and water development. Immediate protective measures are urgently needed to ensure the persistence of finless porpoises in the Yangtze River. The survey design and analytical methods developed in this study might be appropriate for surveys of cetaceans in other river systems. (c) 2008 Elsevier Ltd. All rights reserved.

Effects of Cd2+ and Pb2+ on the substrate biofilms in the integrated vertical-flow constructed wetland

The effects of single Cd2+ and Pb2+, and combined Cd2+ and Pb2+ on dehydrogenase activity and polysaccharide content of the substrate biofilms in the integrated vertical-flow constructed wetland (IVCW) were studied. Dehydrogenase activities decreased linearly with the increasing concentrations of Cd2+ and Pb2+ at different times (6, 24, 72, and 120 h). The activities at both 6 and 24 h were significantly higher than that at 72 and 120 h in the case of single and combined treatments. The single Cd2+ and Pb2+ treatments significantly inhibited dehydrogenase activities at concentrations in excess of 20 mu mol/L Cd2+ and 80 mu mol/L Pb2+, respectively. The inhibitory effect of Cd2+ was much greater than that of Pb2+. At the same time, the combined treatment of Cd2+ and Pb2+ Significantly inhibited dehydrogenase activities at all five concentrations studied and the lowest combined concentration was 1.25 mu mol/L Cd2+ and 5 mu mol/L Pb2+. A synergistic effect of Cd2+ and Pb2+ was observed. On the other hand, polysaccharide contents varied unpredictably with the increasing concentrations of Cd2+ and Pb2+ and extended experimental time. There were no significant statistical differences within the range of concentration and time studied, whether singly or in combination. These results implied that the effects of heavy metals on biofilms should be a concern for the operation and maintenance of constructed wetlands.

Effects of nutrients on the substrate biofilms in the integrated vertical-flow constructed wetland

A L9 orthogonal array design involving 3 factors (C6H12O6, KNO3 and NaH2PO4) and 3 levels for each (C6H12O6: 0.2, 0.4 or 0.8 g/L; KNO3: 0.4, 0.8 or 1.6 g/L, NaH2PO4: 0.05, 0.1 or 0.2 g/L), was used to study the effects of nutrients on dehydrogenase activity and polysaccharide content of substrate biofilms in the integrated vertical-flow constructed wetland (IVCW). Results showed that C6H12O6 and KNO3 were the main factors for dehydrogenase activity and polysaccharide content of biofilms, respectively. The combinations of three nutrients at different concentrations had different effects on dehydrogenase activity and polysaccharide content of biofilms. The optimal combination for dehydrogenase activity was obtained by locating the concentrations Of C6H12O6, KNO3 and NaH2PO4 at 0.2, 0.8 and 0.05 g, and the optimal combination for polysaccharide content was obtained by locating the concentrations Of C6H12O6, KNO3 and NaH2PO4 at 0.2, 0.4 and 0.2 g/L, respectively. The corresponding maximum activity and polysaccharide content were 5.40 mu g TF/g substrate/12 h and 3454.6 mu g/g substrate, respectively. These results would provide the laboratory foundation for optimizing the purification function of the wetland systems.

Differentiate performance of eight filter media in vertical flow constructed wetland: Removal of organic matter, nitrogen and phosphorus

A comparative study was conducted to reveal the differentiate effects of eight different filter media including gravel, zeolites, anthracite, shale, vermiculite, ceramic filter media, blast furnace steel slag and round ceramsite. The study mainly related to the eight different filter media's removal performances of organic matter, nitrogen and phosphorus in the vertical flow constructed wetland simulated system, which treating wastewater at hydraulic loading rate of 1000-2500 mm/d. The results indicated that the removal effects were closely related to the physical and chemical properties of medium materials. Anthracite-filled system had the highest removal rate for the total organic carbon (TOC), up to 70%, and the removal rates of other systems ranged from 20% to 30%. As for the five-day biochemical oxygen demand (BOD5), anthracite-filled and steel slag-filled systems had the highest removal rates, also up to 70%, as well as other systems all exceeded 50%. At the same time, for the total nitrogen (TN) and NH4(+)-N, the zeolites-filled and ceramic-filled systems had the best performances with the removal rates of more than 70%, the other way round, the removal rates of other systems were only about 20%. The distinguishable effects were also observed in removal performances of total phosphorus (TP) and total dissoluble phosphorus (TDP). The removal rates of TP and TDP in steel slag-filled systems were more than 90%, a much higher value, followed by that of the anthracite-filled system, more than 60%, but those of other systems being the less. Our study provided a potential mechanism to optimize the filter media design for the vertical flow constructed wetlands.

Performance and mechanism of phosphorus removal in an integrated vertical flow constructed wetland treating eutrophic lake water

Phosphorus removal performance and a possible mechanism for the phosphorus removal from an eutrophic lake water were investigated using a medium-scale integrated vertical constructed wetland (combined vertical and reverse-vertical systems) from April, 11, 2001 to September, 28, 2004. Environmental factors affecting phosphorus removal and release profiles were monitored simultaneously under hydraulic loads from 400 to 2000 mm per day. The phosphorus removal rate varied with the environmental conditions. The removal rate for acidic influent water was superior to that for alkaline influent water. The substrate in the wetland chamber acted as a buffer to regulate the pH value of the water sample. As regards the water temperature, no significant differences were observed for the removal rate of total phosphorus (TP) and soluble reactive phosphorus (SRP) between low (lower than 15 degrees C) medium (16-25 degrees C) and high temperature (higher than 26 degrees C) conditions. Under a hydraulic load of 400 mm per day, the removal rate reached over 70%, the highest value achieved in this work. In addition, the highest hydraulic load of 2000 mm/d did not result in the lowest removal rate, as had been expected. After a two-year high hydraulic load test, the removal rate decreased significantly. Phosphorous release from the substrate was examined using a spatial sampling method. Depth profiles of total phosphorus and different states of phosphorus present in the substrate were recorded. This further study demonstrated that binding of phosphorus by iron and calcium might be another major factor in the removal and release of TP and SRP in this wetland system. The distribution of the speciated phosphorus showed that the amount of phosphorus captured in the substrate of the down-flow chamber was significantly higher than that captured in the up-flow chamber, suggesting that the up-flow chamber was the main source of phosphorus release in this constructed wetland.

The distribution and economic losses of alien species invasion to China

Invasive alien species have become one of the most serious environmental issues in the world. Data of taxon, origin, pathway, and environmental impacts of invasive alien microorganisms, invertebrates, amphibians and reptiles, fish, birds, mammals, herbs, trees, and, marine organisms in terrestrial, aquatic, and marine ecosystems of China were analyzed during 2001 and 2003, based on literature retrieval and field survey. There were 283 invasive alien species in China, and the number of species of invasive alien microorganisms, aquatic plants, terrestrial plants, aquatic invertebrates, terrestrial invertebrates, amphibians and reptiles, fish, and mammals were 19, 18, 170, 25, 33, 3, 10, and 5, respectively. The proportion of invasive alien species originated from America, Europe, Asia, Africa, and Oceania were 55.1, 21.7, 9.9, 8.1, and 0.6%, respectively. Methods for estimation of direct economic losses to agriculture, forestry, stockbreeding, fishery, road and water transportation, storage, water conservancy, environment and public facilities, and human health were established. Methods for estimation of indirect economic losses caused by invasive alien species to service functions of forest ecosystems, agricultural ecosystems, grassland ecosystems, and wetland ecosystems were also established. The total economic losses caused by invasive alien species to China were to the time of USD 14.45 billion, with direct and indirect economic losses accounting for 16.59% and 83.41% of total economic losses, respectively.

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

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

Population genetic structure of an endemic cyprinid fish, Ancherythroculter nigrocauda, in the upper reaches of the Yangtze River and its implication for conservation

Ancherythroculter nigrocauda is a cyprinid fish endemic to the upper reaches of the Yangtze River, which has been reported to have 2 or 3 chambers to its air bladder. Morphological studies showed no differences between individuals with different types of air bladder, but did demonstrate geographical differences from different sources. After the completion of the Three Gorges Dam, it was expected that the population of this species would decrease, but artificial breeding and stocking is under consideration to protect this species from extinction. In the present study, mtDNA cytochrome b gene sequences were determined and analyzed for A. nigrocauda samples of different morphotypes and sources to identify their genetic differentiations, and thereby guide plans for the artificial propagation and conservation of this species. Haplotype diversity index values (h) and nucleotide diversity values (pi) for all the populations were found to be high indicating their high level genetic diversity. An analysis of molecular variance identified no differentiation among the studied populations. Therefore, we suggested that the individuals of different morphological types and geographical sources belong to the same species. To maintain its high level genetic diversity, it mill he necessary to use large and diverse sources of parental fish for artificial reproduction.

Ex situ conservation status of an endangered Yangtze finless porpoise population (Neophocaena phocaenoides asiaeorientalis) as measured from microsatellites and mtDNA diversity

The Yangtze finless porpoise, Neophocaena phocaenoides asiaeorientalis, is all endangered small cetacean that occurs only in the middle and lower reaches of the Yangtze River of China. The establishment of a breeding population of the porpoise in Tian-e-Zhou Baiji National Natural Reserve represents the first attempt at ex situ conservation efforts for a cetacean species. With the goal of effective protection, management, and monitoring of this preserved population, we examined its genetic diversity using 930 bp of mtDNA control region sequences and 13 polymorphic microsatellite loci. A very low level of genetic variation (h = 0.6010 +/- 0.0029 s.d.; pi = 0.0007 +/- 0.0000002 s.d.) in the mtDNA control region sequences and a moderate genetic diversity (Ho = 0.5740 +/- 0.2575 s.d.) in the microsatellites were detected in the population. It is necessary to introduce more individuals with representative genetic variations into the reserve ill order to foml a larger and healthier group structure for long-term survival of the population. Successful establishment of the Yangtze finless porpoise population in the Reserve also provides a useful model for an ex situ conservation programme for other rare and endangered species. (c) 2005 International Council for the Exploration ofthe Sea. Published by Elsevier Ltd. All rights reserved.

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.