As the most widely used plasticizers in the world, phthalate esters （PAEs） are potential endocrine disruption compounds （EDCs）. In the present study, the toxicity of dimethyl phthalate （DMP）, diethyl phthalate ...As the most widely used plasticizers in the world, phthalate esters （PAEs） are potential endocrine disruption compounds （EDCs）. In the present study, the toxicity of dimethyl phthalate （DMP）, diethyl phthalate （DEP）, dibutyl phthalate （DBP）, di （2-ethylhexyl） phthalate （DEHP） on embryogenesis and larvae development of the marine univalve Haliotis diversicolor supertexta was examined in laboratory. The results show that the malformation of embryos appeared during the experiment, such as embryos died or lysed, small transparent flocculent rings studded on the periphery of the embryo, and the larvae could failed to hatch. In embryo toxic test, embryos incubated at the highest concentration of DMR DEP and DBP solutions showed significantly high abnormal rate compared with the control, while DEHP solutions displayed no significant difference. In larval toxic test, in all concentrations of DMP, DEP and DBP solutions, larval settlement rates were low significantly than that of the control. Similarly, DEHP solutions show nearly no effect on the larval settlement. The order of toxicity on embryos and larvae is DBP〉DEP〉DMP〉DEHE Being a simple and easy stimulation to indoor spawn, sensitive to environmental factors, and short culture time, the embryos of H. diversicolor supertexta can be used to indicate toxicity of the PAEs.展开更多
Using the exposure simulation experiment, the action of petroleum affecting the accumulation of the trace metals including copper （Cu） and cadmium （Cd） in littoral polychaete Nereis diversicolor collected from the...Using the exposure simulation experiment, the action of petroleum affecting the accumulation of the trace metals including copper （Cu） and cadmium （Cd） in littoral polychaete Nereis diversicolor collected from the Shuangtaizi Estuary in Liaoning Province, China was examined. The results showed that there was a markedly non-linear relationship between the accumulation of Cu in worms and the experimental concentration of Cu in exposure solutions when the concentration of petroleum remained at 0, 100, and 220 μl/L, respectively. However, significantly non-linear relationship for worms exposed to Cd was observed only when the concentration of added petroleum was 0 and 220 μl/L. The accumulation of Cu in worms did not differ significantly among the three different levels of petroleum concentrations combined with various concentrations of Cu. So was the accumulation of Cd in worms （p〉0.05）. However, the addition of petroleum in exposure solutions brought about an increase in the accumulation of Cu in Nereis diversicolor, in comparison with single Cu pollution. On the other hand, when the concentration of added petroleum remained at 100 μl/L, the accumulation of Cd in worms was lower than that in worms exposed to various concentrations of only cadmium. However, the worms exposed to Cd and petroleum 220 μl/L did not show obvious and identical increase in the accumulation of Cd, compared with single Cd exposure. The accumulation of both Cu and Cd in worms did not increase significantly with the increases in concentrations of Cu or Cd in exposure solutions combined with petroleum （0, 100, and 220μl/L） under the experimental conditions. Although Nereis diversicolor is exposed to very high Cu and Cd in exposure solutions, accumulation and detoxification mechanisms are sufficient to cope with the extra metal influx in order to survive.展开更多
Ferritin is an iron storage protein that plays a key role in the processes of physiology and pathology.In the present study,the authors reported the ferritin gene from abalone Haliotis diversicolor supertexta,which we...Ferritin is an iron storage protein that plays a key role in the processes of physiology and pathology.In the present study,the authors reported the ferritin gene from abalone Haliotis diversicolor supertexta,which we named hds-ferritin.The full-length of hds-ferritin cDNA consisted of 879 bp with an ORF encoding a 171 amino acids.Amino acid sequence analysis revealed that hds-ferritin shared highly homology with other species.Real time PCR and western blot analysis showed that hds-ferritin was distributed ubiquitously in abalone tissues and had the highest expression level in digestive glands,but its transcripts are not modified remarkably by the stimulation with LPS.The recombinant protein was successfully expressed in Escherichia coli BL21 (DE3),and the titre of anti-ferritin antibody was about 1∶14000.The effects of ROS and RNS on ferritin were analyzed in the present study.The results showed that H2O2 played an important role in decreasing hds-ferritin,however NO cation appeared to have a protecting effect on H2O2-medied reduction of hds-ferritin.展开更多
Abalone Haliotis diversicolor supertexta is an important economic mollusk.The settlement and metamorphosis are two critical stages during its development period,which has direct influence on abalone survival and produ...Abalone Haliotis diversicolor supertexta is an important economic mollusk.The settlement and metamorphosis are two critical stages during its development period,which has direct influence on abalone survival and production.The influence of reactive oxygen species(hydrogen peroxide) on abalone embryo and juvenile development were examined in this study.Larvae of Haliotis diversicolor supertexta were induced to settlement and metamorphose by exposure to seawater supplemented with hydrogen peroxide.They had the best performance at 800 μmol/L.The concentration of 1 000 μmol/L or higher was toxic to the larvae,as the larvae could settle down only at benthic diatom plates without complete metamorphosis.In addition,H2O2 adding time was critical to the larval performance.24h after two-day post-fertilization was proved to be the optimal adding time.In this paper,two action mechanisms of hydrogen peroxide are discussed:(1) hydrogen peroxide has direct toxicity to ciliated cells,thus cause apoptosis;(2) hydrogen peroxide,as a product from catecholamines' autoxidation process in vivo,can reverse this process to produce neuro-transmitters to induce abalone metamorphosis.展开更多
Benthic diatoms constitute the primary diet of abalone during their early stages of development. To evaluate the dietary preferences of early post-larval abalone, Haliotis diversicolor supertexta, we analyzed the gut ...Benthic diatoms constitute the primary diet of abalone during their early stages of development. To evaluate the dietary preferences of early post-larval abalone, Haliotis diversicolor supertexta, we analyzed the gut contents of post-larvae that settled on diatom films. We compared the abundance and species diversity of diatom assemblages in the gut to those of the epiphytic diatom assemblages on the attachment films, and identified 40 benthic diatom species in the gut contents of post-larvae 12 to 24 d after settlement. The most abtmdant taxa in the gut contents were Navicula spp., Amphora copulate, and Amphora coffeaeformis. Navicula spp. accounted for 64.0% of the cell density. In the attachment films, we identified 110 diatom species belonging to 38 genera. Pennate diatoms were the dominant members including the species Amphiprora alata, Cocconeis placentula var. euglypta, Cylindrotheca closterium, Navicula sp. 2, and A. coffeaeformis. Nano-diatoms （〈20 pm in length） accounted for a considerable proportion of the total species number and cell density of the diatom assemblages in the gut contents and on the films. This suggests that nano-diatoms are important to the efficient production of abalone seed. The difference of the composition and abundance of diatoms between in the guts and on the biofilms suggests that early post-larval grazing was selective. An early post-larval abalone preferred nano-diatoms and the genera Navicula and Amphora during the month after settlement.展开更多
The settlement substrates of nona-porous abalones （Haliotis diversicolor supertexta） are covered with biofilms in which several types of microorganisms coexist and interact. These microorganisms are usually importan...The settlement substrates of nona-porous abalones （Haliotis diversicolor supertexta） are covered with biofilms in which several types of microorganisms coexist and interact. These microorganisms are usually important causes of juvenile abalone disease as well as organisms useful in promoting abalones’ adhesion. The bacterial community structure of the biofilms remains unclear. The aim of this research was to determine the genetic diversity and phylogenetic affiliation of the biofilm bacteria. Total DNA of bacteria in biofilms was extracted, and 16S rRNA gene clone library was constructed using the primers specific for the domain bacteria. Subsequently, 30 randomly selected positive clones were screened by PCR-restriction fragment length polymorphism （PCR-RFLP） analysis, and resulted in 15 different RFLP patterns. Sequences analysis of representatives from each unique RFLP type revealed high genetic diversity in the bacterial populations. These sequences fell into nine major lineages of the bacterial domains： α-, β-, γ-and δ-subdivisions of the Proteobacteria; Planctomycete, Actinobacteria, Firmicutes, V errucomicrobium spp., and CytophagaFlexibacter-Bacteroides spp. Phylogenetic analysis indicated that the dominant phylotypes were most closely related to environmental and clinical Burkholderia cepacia of the β-Proteobacteria, and Roseobacteria of the α-Proteobacteria.展开更多
We investigated the effect of tributyltin (TBT) exposure on the concentration of malondialdehyde (MDA) and the activity levels of the superoxide dismutase (SOD),catalase (CAT),and acid and alkaline phosphatase (ACP an...We investigated the effect of tributyltin (TBT) exposure on the concentration of malondialdehyde (MDA) and the activity levels of the superoxide dismutase (SOD),catalase (CAT),and acid and alkaline phosphatase (ACP and AKP) enzymes in the small abalone,Haliotis diversicolor supertexta.We collected samples of the hepatopancreas and hemolymph 2,6,24,48,96,and 192 h after exposure to 0.35 μg (Sn)/L TBT.In the hepatopancreas,ACP activity was significantly higher in animals exposed to TBT 2,24,and 96 h post-exposure compared with the control animals.AKP activity was also higher after 2 h,but SOD and CAT activity was unchanged.The concentration of MDA in the hemolymph was significantly higher than the control animals 2 and 6 h post-exposure.In the hemolymph of animals exposed to TBT,ACP activity was significantly lower than in the control animals 192 h post-exposure,whereas AKP activity was significantly lower 2 and 192 h post-exposure.Hemolymph SOD activity and levels of MDA were significantly lower than in the control animals 24 h after exposure but significantly higher after 96 h.Our results demonstrate that exposure to TBT cause rapid changes in ACP and AKP activity as well as altering the concentration of MDA in the hepatopancreas and hemolymph.SOD and CAT do not appear to be involved in the detoxification of TBT in the hepatopancreas of small abalone.展开更多
Haliotis diversicolor Reeve is one of the most important mollusks cultured in South China. Preliminary genetic linkage maps were constructed with amplified fragment length polymorphism (AFLP) markers. A total of 2 596...Haliotis diversicolor Reeve is one of the most important mollusks cultured in South China. Preliminary genetic linkage maps were constructed with amplified fragment length polymorphism (AFLP) markers. A total of 2 596 AFLP markers were obtained from 28 primer combinations in two parents and 78 offsprings. Among them, 412 markers (15.9%) were polymorphic and segregated in the mapping family. Chi-square tests showed that 151 (84.4%) markers segregated according to the expected 1:1 Mendelian ratio (P<0.05) in the female parent, and 200 (85.8%) in the male parent. For the female map, 179 markers were used for linkage analysis and 90 markers were assigned to 17 linkage groups with an average interval length of 25.7 cm. For the male map, 233 markers were used and 94 were mapped into 18 linkage groups, with an average interval of 25.0 cm. The estimated genome length was 2 773.0 cm for the female and 2 817.1 cm for the male map. The observed length of the linkage map was 1 875.2 cm and 1 896.5 cm for the female and male maps, respectively. When doublets were considered, the map length increased to 2 152.8 cm for the female and 2 032.7 cm for the male map, corresponding to genome coverage of 77.6% and 72.2%, respectively.展开更多
Fine structure of the hypobranchial gland of Haliotis diversicolor has been studied by light and electron microscopy. The results showed that the gland is folded pleats organ, which is highly glandular area of the epi...Fine structure of the hypobranchial gland of Haliotis diversicolor has been studied by light and electron microscopy. The results showed that the gland is folded pleats organ, which is highly glandular area of the epidermal lining the roof of the mantle cavity. Two such glandular areas, one on each side of the rectum, occur in H. diversicolor. Left one is much larger than fight one. By light microscope, on H-E stained section, four cell types can be divided： cells with weak basophilic fibrillar elements; with acidophilic granular substance; with strong basophiIic fibrillar elements and ciliated ceils. In the basal lamina region under the gland epithelium, there are a few connective tissues. Surface view of the gland could be seen by scanning electron microscope, there are cilia and different kinds of secretions distributed. By transmission electron microscope, supporting cells, sensory cells and seven types gland cells were observed to form the glandular epithelium. Cells are rich in rough endoplasmic reticulum, smooth muscle fiber and nerve endings were found beneath glandular epithelium, between basal lamina.展开更多
基金Supported by National Natural Science Foundation of China (No. 40476057)
文摘As the most widely used plasticizers in the world, phthalate esters （PAEs） are potential endocrine disruption compounds （EDCs）. In the present study, the toxicity of dimethyl phthalate （DMP）, diethyl phthalate （DEP）, dibutyl phthalate （DBP）, di （2-ethylhexyl） phthalate （DEHP） on embryogenesis and larvae development of the marine univalve Haliotis diversicolor supertexta was examined in laboratory. The results show that the malformation of embryos appeared during the experiment, such as embryos died or lysed, small transparent flocculent rings studded on the periphery of the embryo, and the larvae could failed to hatch. In embryo toxic test, embryos incubated at the highest concentration of DMR DEP and DBP solutions showed significantly high abnormal rate compared with the control, while DEHP solutions displayed no significant difference. In larval toxic test, in all concentrations of DMP, DEP and DBP solutions, larval settlement rates were low significantly than that of the control. Similarly, DEHP solutions show nearly no effect on the larval settlement. The order of toxicity on embryos and larvae is DBP〉DEP〉DMP〉DEHE Being a simple and easy stimulation to indoor spawn, sensitive to environmental factors, and short culture time, the embryos of H. diversicolor supertexta can be used to indicate toxicity of the PAEs.
基金The National Natural Science Foundation of China （No. 20477049） and the Key Basic Research Development and Planning Project （No. 2004CB418503）
文摘Using the exposure simulation experiment, the action of petroleum affecting the accumulation of the trace metals including copper （Cu） and cadmium （Cd） in littoral polychaete Nereis diversicolor collected from the Shuangtaizi Estuary in Liaoning Province, China was examined. The results showed that there was a markedly non-linear relationship between the accumulation of Cu in worms and the experimental concentration of Cu in exposure solutions when the concentration of petroleum remained at 0, 100, and 220 μl/L, respectively. However, significantly non-linear relationship for worms exposed to Cd was observed only when the concentration of added petroleum was 0 and 220 μl/L. The accumulation of Cu in worms did not differ significantly among the three different levels of petroleum concentrations combined with various concentrations of Cu. So was the accumulation of Cd in worms （p〉0.05）. However, the addition of petroleum in exposure solutions brought about an increase in the accumulation of Cu in Nereis diversicolor, in comparison with single Cu pollution. On the other hand, when the concentration of added petroleum remained at 100 μl/L, the accumulation of Cd in worms was lower than that in worms exposed to various concentrations of only cadmium. However, the worms exposed to Cd and petroleum 220 μl/L did not show obvious and identical increase in the accumulation of Cd, compared with single Cd exposure. The accumulation of both Cu and Cd in worms did not increase significantly with the increases in concentrations of Cu or Cd in exposure solutions combined with petroleum （0, 100, and 220μl/L） under the experimental conditions. Although Nereis diversicolor is exposed to very high Cu and Cd in exposure solutions, accumulation and detoxification mechanisms are sufficient to cope with the extra metal influx in order to survive.
基金The National Natural Science Foundation of China under contract No. 30671619the Natural Science Foundation of Zhejiang Province under contract No. Y307606
文摘Ferritin is an iron storage protein that plays a key role in the processes of physiology and pathology.In the present study,the authors reported the ferritin gene from abalone Haliotis diversicolor supertexta,which we named hds-ferritin.The full-length of hds-ferritin cDNA consisted of 879 bp with an ORF encoding a 171 amino acids.Amino acid sequence analysis revealed that hds-ferritin shared highly homology with other species.Real time PCR and western blot analysis showed that hds-ferritin was distributed ubiquitously in abalone tissues and had the highest expression level in digestive glands,but its transcripts are not modified remarkably by the stimulation with LPS.The recombinant protein was successfully expressed in Escherichia coli BL21 (DE3),and the titre of anti-ferritin antibody was about 1∶14000.The effects of ROS and RNS on ferritin were analyzed in the present study.The results showed that H2O2 played an important role in decreasing hds-ferritin,however NO cation appeared to have a protecting effect on H2O2-medied reduction of hds-ferritin.
基金Supported by NSFC (No. 40476057)Shenzhen Science and Technology Foundation (No. JH200504270120A)the Longgang Aquaculture Bureau of Shenzhen
文摘Abalone Haliotis diversicolor supertexta is an important economic mollusk.The settlement and metamorphosis are two critical stages during its development period,which has direct influence on abalone survival and production.The influence of reactive oxygen species(hydrogen peroxide) on abalone embryo and juvenile development were examined in this study.Larvae of Haliotis diversicolor supertexta were induced to settlement and metamorphose by exposure to seawater supplemented with hydrogen peroxide.They had the best performance at 800 μmol/L.The concentration of 1 000 μmol/L or higher was toxic to the larvae,as the larvae could settle down only at benthic diatom plates without complete metamorphosis.In addition,H2O2 adding time was critical to the larval performance.24h after two-day post-fertilization was proved to be the optimal adding time.In this paper,two action mechanisms of hydrogen peroxide are discussed:(1) hydrogen peroxide has direct toxicity to ciliated cells,thus cause apoptosis;(2) hydrogen peroxide,as a product from catecholamines' autoxidation process in vivo,can reverse this process to produce neuro-transmitters to induce abalone metamorphosis.
基金Supported by Special Project on Basic Research of China,Ministry of Science and Technology (No 2006FY120100)Natural Science Foundation of Fujian Province (No B05100052005YZ1024)
文摘Benthic diatoms constitute the primary diet of abalone during their early stages of development. To evaluate the dietary preferences of early post-larval abalone, Haliotis diversicolor supertexta, we analyzed the gut contents of post-larvae that settled on diatom films. We compared the abundance and species diversity of diatom assemblages in the gut to those of the epiphytic diatom assemblages on the attachment films, and identified 40 benthic diatom species in the gut contents of post-larvae 12 to 24 d after settlement. The most abtmdant taxa in the gut contents were Navicula spp., Amphora copulate, and Amphora coffeaeformis. Navicula spp. accounted for 64.0% of the cell density. In the attachment films, we identified 110 diatom species belonging to 38 genera. Pennate diatoms were the dominant members including the species Amphiprora alata, Cocconeis placentula var. euglypta, Cylindrotheca closterium, Navicula sp. 2, and A. coffeaeformis. Nano-diatoms （〈20 pm in length） accounted for a considerable proportion of the total species number and cell density of the diatom assemblages in the gut contents and on the films. This suggests that nano-diatoms are important to the efficient production of abalone seed. The difference of the composition and abundance of diatoms between in the guts and on the biofilms suggests that early post-larval grazing was selective. An early post-larval abalone preferred nano-diatoms and the genera Navicula and Amphora during the month after settlement.
基金The Natural Science Foundation of Fujian Province of China under contract Nos 2005k055 and B0740012the Innovative Research Team under contract No 2006A001Science Foundation of Jimei University, China
文摘The settlement substrates of nona-porous abalones （Haliotis diversicolor supertexta） are covered with biofilms in which several types of microorganisms coexist and interact. These microorganisms are usually important causes of juvenile abalone disease as well as organisms useful in promoting abalones’ adhesion. The bacterial community structure of the biofilms remains unclear. The aim of this research was to determine the genetic diversity and phylogenetic affiliation of the biofilm bacteria. Total DNA of bacteria in biofilms was extracted, and 16S rRNA gene clone library was constructed using the primers specific for the domain bacteria. Subsequently, 30 randomly selected positive clones were screened by PCR-restriction fragment length polymorphism （PCR-RFLP） analysis, and resulted in 15 different RFLP patterns. Sequences analysis of representatives from each unique RFLP type revealed high genetic diversity in the bacterial populations. These sequences fell into nine major lineages of the bacterial domains： α-, β-, γ-and δ-subdivisions of the Proteobacteria; Planctomycete, Actinobacteria, Firmicutes, V errucomicrobium spp., and CytophagaFlexibacter-Bacteroides spp. Phylogenetic analysis indicated that the dominant phylotypes were most closely related to environmental and clinical Burkholderia cepacia of the β-Proteobacteria, and Roseobacteria of the α-Proteobacteria.
基金Supported by the Committee of Xiamen Science and Technology,Xiamen,China (No.502Z20055024)the National Natural Science Foundation of China (No.20877034)the Innovation Team Foundation of Jimei University (No.2008A001)
文摘We investigated the effect of tributyltin (TBT) exposure on the concentration of malondialdehyde (MDA) and the activity levels of the superoxide dismutase (SOD),catalase (CAT),and acid and alkaline phosphatase (ACP and AKP) enzymes in the small abalone,Haliotis diversicolor supertexta.We collected samples of the hepatopancreas and hemolymph 2,6,24,48,96,and 192 h after exposure to 0.35 μg (Sn)/L TBT.In the hepatopancreas,ACP activity was significantly higher in animals exposed to TBT 2,24,and 96 h post-exposure compared with the control animals.AKP activity was also higher after 2 h,but SOD and CAT activity was unchanged.The concentration of MDA in the hemolymph was significantly higher than the control animals 2 and 6 h post-exposure.In the hemolymph of animals exposed to TBT,ACP activity was significantly lower than in the control animals 192 h post-exposure,whereas AKP activity was significantly lower 2 and 192 h post-exposure.Hemolymph SOD activity and levels of MDA were significantly lower than in the control animals 24 h after exposure but significantly higher after 96 h.Our results demonstrate that exposure to TBT cause rapid changes in ACP and AKP activity as well as altering the concentration of MDA in the hepatopancreas and hemolymph.SOD and CAT do not appear to be involved in the detoxification of TBT in the hepatopancreas of small abalone.
基金Supported by the National Basic Research Program of China (973 Program) (Nos. 2010CB126405 and 2009CB126005)the Key Project for International Science and Technology Cooperation, Ministry of Science and Technology of China (No. 2004DFA07200)+2 种基金the Great Science and Technique Program of Hainan Province (No. 06120)the Hainan Key Laboratory of Tropical Hydrobiological Technology (No. shkyjj0810)supported by the Rutgers University
文摘Haliotis diversicolor Reeve is one of the most important mollusks cultured in South China. Preliminary genetic linkage maps were constructed with amplified fragment length polymorphism (AFLP) markers. A total of 2 596 AFLP markers were obtained from 28 primer combinations in two parents and 78 offsprings. Among them, 412 markers (15.9%) were polymorphic and segregated in the mapping family. Chi-square tests showed that 151 (84.4%) markers segregated according to the expected 1:1 Mendelian ratio (P<0.05) in the female parent, and 200 (85.8%) in the male parent. For the female map, 179 markers were used for linkage analysis and 90 markers were assigned to 17 linkage groups with an average interval length of 25.7 cm. For the male map, 233 markers were used and 94 were mapped into 18 linkage groups, with an average interval of 25.0 cm. The estimated genome length was 2 773.0 cm for the female and 2 817.1 cm for the male map. The observed length of the linkage map was 1 875.2 cm and 1 896.5 cm for the female and male maps, respectively. When doublets were considered, the map length increased to 2 152.8 cm for the female and 2 032.7 cm for the male map, corresponding to genome coverage of 77.6% and 72.2%, respectively.
基金Acknowledgments： This study was founded by Natural Science Foundation of Guangdong （No. 032248） and Large Instrument Using Fund of South China Agricultural University （ No. 2007Y002 ） .
文摘Fine structure of the hypobranchial gland of Haliotis diversicolor has been studied by light and electron microscopy. The results showed that the gland is folded pleats organ, which is highly glandular area of the epidermal lining the roof of the mantle cavity. Two such glandular areas, one on each side of the rectum, occur in H. diversicolor. Left one is much larger than fight one. By light microscope, on H-E stained section, four cell types can be divided： cells with weak basophilic fibrillar elements; with acidophilic granular substance; with strong basophiIic fibrillar elements and ciliated ceils. In the basal lamina region under the gland epithelium, there are a few connective tissues. Surface view of the gland could be seen by scanning electron microscope, there are cilia and different kinds of secretions distributed. By transmission electron microscope, supporting cells, sensory cells and seven types gland cells were observed to form the glandular epithelium. Cells are rich in rough endoplasmic reticulum, smooth muscle fiber and nerve endings were found beneath glandular epithelium, between basal lamina.
