Estrogen alters the profile of the transcriptome in river snail Bellamya aeruginosa.

We evaluated the transcriptome dynamics of the freshwater river snail Bellamya aeruginosa exposed to 17ß-estradiol (E2) using the Roche/454 GS-FLX platform. In total, 41,869 unigenes, with an average length of 586 bp, representing 36,181 contigs and 5,688 singlets were obtained. Among them, 18.08, 36.85, and 25.47 % matched sequences in the GenBank non-redundant nucleic acid database, non-redundant protein database, and Swiss protein database, respectively. Annotation of the unigenes with gene ontology, and then mapping them to biological pathways, revealed large groups of genes related to growth, development, reproduction, signal transduction, and defense mechanisms. Significant differences were found in gene expression in both liver and testicular tissues between control and E2-exposed organisms. These changes in gene expression will help in understanding the molecular mechanisms of the response to physiological stress in the river snail exposed to estrogen, and will facilitate research into biological processes and underlying physiological adaptations to xenoestrogen exposure in gastropods.

[Research Progress on Characteristics of Human Microplastic Pollution and Health Risks].

Microplastic pollution is not only an environmental problem but also a social problem. Many studies have been conducted on the sources, abundance, and distribution of microplastics in the environment, but an understanding of human exposure levels and potential health risks remains very limited. Based on the bibliometric methods, the present review systematically summarized the exposure pathways of microplastics in humans, and then the characteristics and potential adverse impacts on human health were expounded upon. Available literature showed that microplastics in human bodies were mainly concentrated on sizes smaller than 50 µm, and polyethylene (PE), polypropylene (PP), and polyethylene terephthalate (PET) were the main polymers. Microplastics in environments entered human bodies mainly through food and respiratory pathways, then accumulated in lung and gastrointestinal tissues. Most importantly, small-sized microplastics could distribute in tissues and organs via the circulatory system. The results from lab-based toxicological experiments showed that microplastics not only posed threats to cell membrane integrity, immune stress, gut microbiota, and energy metabolism but also had potentially adverse impacts on the reproductive system. To further understand the health risks of microplastic pollution, it is necessary to promote research on the toxicological effects of microplastics as well as the inner mechanisms and also to establish risk assessment frameworks for evaluating microplastic pollution. These works are crucial to preventing the risks of microplastic pollution with scientific evidence.

Imposex effects on the veined rapa whelk (Rapana venosa) in Bohai Bay, China.

Environmentally relevant concentrations of organotin compounds (OTs) may trigger sex changes in marine invertebrates and pose a threat to the marine ecosystem. In this study, we investigated organotin levels and the biological responses of wild veined rapa whelk (Rapana venosa) from Lüjuhe district (LJH), Dashentang district (DST), and Nanpaihe district (NPH) in Bohai Bay, China. We found that 11.11 and 22.95 % of the veined rapa whelks from DST and NPH exhibited imposex characteristics with a relative penis size index (RPSI) of 12.50 and 12.31, respectively. The RNA/DNA ratio was significantly lower in females from DST than those from LJH (p < 0.05), and a slight increase in DNA damage was observed in females and imposex individuals compared to males. Moreover, less genetic distance occurred between LJH and NPH (0.016) than between LJH and DST (0.028), although they belonged to the same regional population. OTs analysis showed that triphenyltin chloride concentrations (41.45 ng/g dried weight) were significantly higher than tributyltin concentrations (9.51 ng/g dried weight) in tissues (p < 0.05), but no significant differences were observed in sediments (p > 0.05). In conclusion, the occurrence of imposex individuals and biological responses of the wild veined rapa whelk from Bohai Bay suggest that the marine ecosystem might be at risk.

Construction and characterization of a normalized cDNA library from the river snail Bellamya aeruginosa after exposure to copper.

The construction of a normalized cDNA library is a popular tool for identifying novel biomarkers for monitoring environmental pollution. In the present study, a normalized cDNA library was constructed from the river snail Bellamya aeruginosa after exposure to Cu(2+) by using the SMART technique. The titer of the cDNA library was 1.78 × 10(6) pfu/ml, with a recombinant efficiency of 95.8%. In addition, from 6,000 randomly selected and sequenced clones, 5,473 high-quality ESTs were identified. After processing the sequences, 3,961 unigenes representing 897 contigs and 3,064 singlets were obtained with 27.6% redundancy. Analysis of expressed sequenced tags using COG and GO annotation and KEGG pathway data showed that a large group of genes related to growth and development, signal transduction, and defense mechanisms were present in the cDNA library. Based on our findings, this normalized cDNA library will provide a valuable resource for further research on functional genes and ecotoxicology in B. aeruginosa.

[Policy Analysis in Plastic Pollution Governance and Recommendations in China].

Plastic pollution is a global concern and an issue in environmental governance. Based on the updated "Plastic Prohibition/Restriction Order" issued recently in China, the present study systematically reviewed the implementation effectiveness of the "Plastic Prohibition/Restriction Order" since 2007. Furthermore, we summarized plastics in China and plastic waste management progress policies. Additionally, three deficiencies of the updated "Plastic Prohibition/Restriction Order" were discussed:policy formulation, implementation, and supervision. Some positive recommendations were provided based on the available reports, such as integrating plastic pollution into national basic laws and regulation systems, building a network platform for public attendance, and coordinating the publicity of the "Plastic prohibition/Restriction Order" with the public interest. Besides these measurements, some points about plastic waste management in the future were also highlighted, such as the "blind area" in small retail stores, the rational sharing of environmental protection responsibility, new materials and processes, and recycling and disposal systems for plastic wastes. Most importantly, the present study could provide ideas for policy-makers to address plastic pollution at its sources.

Ribosomal protein L7 as a suitable reference gene for quantifying gene expression in gastropod Bellamya aeruginosa.

Expression levels of eight candidate reference genes were quantified in tissues of gastropod Bellamya aeruginosa exposed for 10 d to various stressors, including fasting, 17ß-estradiol, 17α-methyltestosterone, and Cd(2+). The results showed that 18s rRNA was the most highly expressed of the candidate reference genes, while H2A was the least expressed. There were no significant changes (p>0.05) in the expression of the eight genes in tissues among the different treatments. Using RefFinder to evaluate the expression stabilities of the eight candidate reference genes, ribosomal protein was shown to be the most stable reference gene, and no effects were observed among the different stressor treatments. These results indicate that RPL 7 is the most suitable reference gene for quantifying gene expression in B. aeruginosa under environmental stress, which was verified in B. aeruginosa exposed to high doses of E2 for 24 and 72h.

Use of heat shock protein mRNA expressions as biomarkers in wild crucian carp for monitoring water quality.

In organisms, the production of heat shock proteins (HSPs) can be induced after exposure to various stressors. In the present study, partial cDNA sequences encoding of HSP30, HSP60, HSP70, and HSP90 genes were isolated and sequenced in the crucian carp (Carassius carassius). Next, quantitative real-time reverse transcription PCR (qRT-PCR) assays normalized to beta-actin were developed to quantify HSP mRNA expression. The same methods were used to detect HSP mRNA expression in the wild crucian carp that were collected from the Hun River. In fish located downstream of the river, we found that the levels of HSP70 and HSP30 in the liver and kidney were higher than fish located upstream, and these differences coincided with changes of the water quality. These results suggest that kidney HSP30 and liver HSP70 expression can serve as sensitive biomarkers for the presence of field environmental stressors in wild crucian carp.

Evidence for the presence of sex steroid hormones in Zhikong scallop, Chlamys farreri.

To obtain evidence of the presence of sex steroid hormones in mollusks, hormone variation in the gonads of the Zhikong scallop Chlamys farreri was analyzed using UPLC-MS/MS. These were found, as expected, with concentrations of estrone (E1), 17beta-estradiol (E2), and testosterone (T) in the testes ranging from not detected (ND) to 0.07 ± 0.10, ND to 3.10 ± 2.00, and ND to 2.67 ± 1.55 ng/g wet weight, respectively. In the ovaries, these hormones ranged from ND to 2.45 ± 1.22, ND to 27.90 ± 4.23, and ND to 2.38 ± 1.56 ng/g ww, respectively. The levels of T in males and E2 in females followed a trend similar to the gonadal-somatic index over the course of the reproductive period. In addition, the gene expression of vitellogenin and calmodulin-2 showed similar patterns to T and E2, while the estrogen receptors and calmodulin-1 did not. These results indicate that sex steroids are present in the scallop and that they may regulate endocrine functions during the reproductive process.

Transcriptomic response to estrogen exposure in the male Zhikong scallop, Chlamys farreri.

The transcriptomes of Zhikong scallop exposed to 17ß-estradiol were determined using the Roche/454. A total of 51,997 unigenes, representing 45,030 contigs and 6967 singlets were obtained. And 14,028, 19,798 and 14,981 of these unigenes were annotated from the non-redundant nucleic acid database, non-redundant protein database and Swiss protein database, respectively. A total of 10,699 unigenes were further annotated to biological processes (9080), molecular functions (8692) and cellular components (7829) using the GO, and 8945 unigenes were mapped to biological pathways including the metabolism (2862) and genetic information processing (2263). Most importantly, 16,692 unigenes and 18,686 unigenes in testis, and 10,492 unigenes and 13,186 unigenes in digestive gland were up-regulated significantly after exposure to 50 and 500 ng E2/L; while 10,212 unigenes and 9409 unigenes in testis and 10,629 unigenes and 9463 unigenes in digestive gland were down-regulated. These valuable information provides insights into the mechanisms in invertebrate exposure to EDCs.

[Study on the ecological risk of wild veined rapa whelk (Rapana venosa) exposured to organotin compounds in Bohai Bay, China].

The present study evaluated the potential ecological risk of organotin compounds (OTs) to wild veined rapa whelk (Rapana Venosa) population in Bohai Bay. The results showed that the imposex rate was 12.5% and 6.48% in Dashentang and Nanpaihe coastal areas, with relative penis size index of 9.61 and 12.45, respectively. The concentrations of butyltin compounds and phenyltin compounds were 39.04 ng x g(-1) dw and 46.48 ng x g(-1) dw in muscle tissues, and 32.09 ng x g(-1) dw and 109.03 ng x g(-1) dw in digest gland, respectively. Based on TBT levels in the muscles of all samples, a risk quotient of 0.024 was derived, indicating certain risk of OTs at current levels to wild veined rapa whelk populations in Bohai Bay.

Biomarker responses and genotoxicity in the mud snail (Bullacta exarata) as indicators of coastal contamination.

In the present study different biomarker responses and genotoxicity were determined in wild mud snails (Bullacta exarata) collected from 3 sampling sites in Bohai Bay in northeastern China, which is a region that is under considerable anthropogenic influence. Significant spatial variability of superoxide dismutase (SOD) and metallothionein (MT)-like proteins were recorded, while glutathione transferase (GST), catalase (CAT), and vitellin-like proteins (Vn's) were not observed. Furthermore, genomic DNA that was amplified with 4 fluorescence-labeled primer pairs showed variable genetic distances among the 3 wild mud snail populations found in Bohai Bay, which may be correlated with differences in the types of environmental genotoxicants, such as heavy metals and polycyclic aromatic hydrocarbons. This multi-biomarker approach provided an improved understanding of the potential toxicological impact of contaminated sediments on benthic organisms.

[Characteristics of heavy metal pollution in Daninghe River and Modaoxi River of Three Gorges Reservoir areas].

In order to explore the potential ecological risk of heavy metals in Three Gorges Reservoir areas, the present study analyzed the heavy metals in sediments and wild crucain carp from Daning River and Modaoxi River which are two typical tributaries, and then the potential risk was evaluated using potential ecological risk index for sediments and comprehensive pollution index for fish, respectively. The results showed that concentrations of Zn and Cr were higher (Daning River: 78.31 and 83.98 mg x kg(-1); Modaoxi River: 99.03 and 94.20 mg x kg(-1)), while Cd was the lowest (Daning River: 0.62 mg x kg(-1); Modaoxi River: 0.75 mg x kg(-1)). Moreover, an obvious increasing trend of these elements was observed in these tributaries from upstream to downstream. For the wild crucain carp, the lowest concentrations of these elements were detected in muscles, and the highest concentrations were in the intestinal. However, no obvious increasing trend of these elements was observed from upstream to downstream. The potential ecological risk in sediments showed a high risk, while no risk was observed in fish.