池塘养殖刺参（Apostichopus japonicus）食物来源的稳定同位素法研究
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摘要
本研究以养殖刺参(Apostichopus japonicus)为对象,采用稳定同位素比率法定量计算了刺参在各种环境下的食物来源,为刺参增养殖实践提供了科学的参考资料。
1、为避免海泥用作刺参饲料添加物的负面作用,寻找其替代物是当前刺参规模化养殖需要解决问题之一。本实验拟采用稳定碳同位素比率法研究用黄泥代替海泥的可行性。结果：添加20%黄泥、海泥的绿藻粉分别按7.5%刺参幼参体重投喂60天,刺参的特定生长率SGR分别为(1.95±0.13)%d-1及(1.72±0.17)%d-1,黄泥、海泥对刺参食物来源贡献分别为(9.78±1.37)%、(9.89±1.47)%,绿藻粉对刺参食物来源贡献分别为(90.22±1.37)%、(90.11±1.47)%,以上各项指标在黄泥组刺参和海泥组间均无显著性差异(P>0.05)。结果表明,用黄泥代替海泥完全可行,并能降低养殖成本及对浅海底质环境的破坏。
2、通过分析刺参养殖池塘内采集到的样品的碳稳定同位素值(δ13C值)发现：底栖动物的稳定碳同位素值与其食物来源密切相关,故可用于研究其食物来源组成。刺参的主要食物来源按δ13C值范围大致可划分为3类：即水体中的植物性碳源(主要为底栖硅藻等)、动物性碳源(小型底栖生物等)以及沉积物有机质(底泥有机质SOM)。研究结果表明,稳定碳同位素是分析刺参养殖池塘食物网结构的有效工具。
3、实验采用室内实验与池塘实验相结合方法,池塘实验设计了5,10,15,25,35头/m2的刺参养殖密度,日投饵率5%刺参体重,投喂60d来研究人工饲料对幼刺参生长的贡献。结果：(1)人工饲料对室内水族箱、池塘围隔内刺参的最终体重影响显著(F=148.541,P=0.003;F=6.301,P=0.018),饲料组围隔内刺参的最终体重均高于同密度对照组;放养密度对围隔内刺参的最终体重影响极显著(F=13.009,P<0.01),随密度增大最终体重减小；(2)人工饲料对水族箱刺参、补充饲料与密度及其互作对池塘刺参的最终δ13C影响均显著(F=3.0,P=0.032;F=18.856,P<0.01;F=4.954,P=0.003; F=20.421,P<0.01)。投喂补充饲料的围隔内刺参的δ13C值的随着密度的增大由(-13.262±0.183)‰下降到(-15.102±0.189)‰。(3)补充饲料在不同的密度下对刺参的食物贡献差异均极显著(F=24.439,P<0.01),随着密度增大,人工饲料对刺参食物贡献增加,由最低密度5头/m2时(3.78±2.98)%的食物贡献,增加到最高密度35头/m2时的(29.48±3.31)%。本实验为刺参增养殖实践人工饲料的使用提供了科学的参考。
4、采用碳氮双稳定同位素法分析了池底改造的池塘养殖刺参的食物来源。实验结果表明：实验刺参的特定生长率SGR为1.9%d-1,其成活率为64.7%。应用IsoSource软件计算得出：不同食物来源对刺参生长贡献分别为细菌26.6%(13～34%)、沉积物21.5%(4～40%)、大型藻类19.1%(0～52%)、小型底栖生物18%(0～49%)及硅藻14.8%(0～41%)。可见,刺参为杂食的沉积物食性生物。
5、利用稳定碳同位素比率法定量分析了石头、瓦片、塑料管、空心砖和水泥管这五种附着基为养殖刺参提供的食物来源情况。结果发现,底栖微藻在五种附着基表层物中均是主要碳源(39.9%～73%),而沉积物和细菌分别占7.3%～34.5%与9.7%～29.6%。不同附着基表层物在刺参食物来源中放的贡献平均在14.5%～32.6%范围,由大到小依次为塑料管>空心砖>水泥管>瓦片>石块。本研究结果对刺参规模化养殖中附着基的选择有一定的借鉴作用。
6、利用稳定碳同位素法定量分析了混养栉孔扇贝(Chlamys (Azumapecten)Farreri)对刺参(Apostichopus japonicus)生长及食物来源的影响。结果发现：混养模式下刺参的特定生长率SGR为(1.43±0.72)%d-1、存活率为(96.67±5.77)%,两者均略高于单养模式的(1.29±0.62)%d-1和(93.3±5.77)%,上述指标均无显著性差异(P=0.709；P=0.519)。在单养刺参的食物来源,由贡献率大到小分别为微藻62%、细菌20.6%、沉积物17.4%；在混养刺参的食物来源,按贡献率由大到小分别为微藻51.9%、细菌22%、沉积物15.8%、扇贝沉降物10.3%。本研究结果对刺参混养模式的选择有一定的借鉴作用。
1. How to eliminate negative effects of sea mud in feed for juvenile sea cucumber (Apostichopus japonicus) is an unsolved problem. Here we propose a method that using stable carbon isotope ratio to prove possibilities of replacement for sea mud with yellow soil. The results showed that feeding juveniles with algae contained 20% yellow soil and sea mud at 7.5% their body weights and for 60 days, the SGR of A. japonicus were (1.95±0.13)%, (1.72±0.17)%, contributions of yellow soil and sea mud to food sources of juveniles were (9.78±1.37)% and (9.89±1.47)%, contributions of green algae to food sources of juveniles were(90.22±1.37)% and (90.11±1.47)%, there was no significant difference for the same index between treatments of sea mud with yellow soil(P>0.05). These results support replacing sea mud with yellow soil is of great feasibility and can save costs and minimize the disturbance of Shallow sea beds.
2. By analysing stable carbon isotopic values (δ13C values) of the organisms collected in a sea cucumber farming pond, it is found that the 813C values of one animal has a close relation with that of the digested food and may be useful in elucidating their food sources. Using the stable carbon isotopic tracers may help confirm the importances and compositions of food source for filter feeders and predators. The investigations show that the main food sources of the Apostichopus japonicus can be divided into three groups in terms of stable carbon isotopic composition:benthic diatoms, meiofauna and organic matter in sediments (POM and SOM).The results lead to a conclusion that theδ13C value is useful tracer to investigate the food web structure of coastal ponds.
3. Stable carbon isotope ratios (δ13C) were used to estimate the contribution of supplemental feed to growth of juvenile sea cucumber cultured in pond enclosures. Five different culture densities (5,10,15,25 and 35 individuals /m2) of sea cucumbers were cultured in pond enclosures for 60 days, keeping constant feeding ration of 5% initial weight. The results were as followed:1) supplemental feed had significant effects on final weights of sea cucumbers both in aquarium and enclosure study(F=148.541, P=0.003; F=6.301, P=0.018, final weights sea cucumbers fed on supplemental feed were higher than that in unfed enclosures, significant differences were observed among different densities (F=13.009, P<0.01), a decrease in final weight with increasing stocking density was described.2) Supplemental feed had significant effects onδ13C values of sea cucumbers in aquariums, Supplemental feed, density had significant effects onδ13C values of sea cucumbers in aquariums, Supplemental feed, and density had significant interaction onδ13C values of sea cucumbers in aquariums. Theδ13C values of sea cucumbers fed on supplemental feed showed a descending trend, ranged from (-13.262±0.183)‰to (-15.102±0.189)‰, as the density increased from 5 to 35 inds./m2. It was calculated that a higher contribution of supplemental feed to sea cucumber growth was favored when density was increased, reaching a minimum (3.78±2.98)% at 5 inds./m2 and a maximum (29.48±3.31)% under the highest stocking condition.The contributions of supplemental feed showed significant differences among different densities (F=24.439, P<0.01). Food limitation may account for a fact of lower final weights while higher feed contributions were obtained under high density conditions.Results from this work should be an aid to developing intensive culture and optimum feed for holothurians.
4. Food sources of sea cucumber cultured in a bottom-renovated pond were determined using stable carbon and nitrogen isotope biomarkers. Based on computing result of the IsoSource software, Apostichopus japonicus's diet consisted of bacteria(13%～34%), sediments(4%～40%), diatoms(0～41%), macroalgae(0～52%), and meiofauna(0%～49%). Playing a primary role in the feeding of sea cucumber, bacteria contributed average 26.6% possible food sources, sort by size, contributions of sediments, macroalgae, meiofauna and diatom were 21.5%,19.1%,18%,14.8%, respectively.
5. Food contribution of stones, tiles, plasic pipes, hollow bricks and cement pipes to cultured sea cucumber (Apostichopus japonicus a) in enclosures were determined using stable carbon isotope biomarker.The surface organic matter of five settlement substrata consisted of diatoms, sediments, bacteria and bacteria. Playing a primary role in the surface organic matter, diatom contributed verage 39.9%～73% possible food sources, sort by size, contribution of sediments and bacteria were 17.3%～34.5% and 9.7%～29.6%, respectively. Contributions of the surface organic matter of five settlement substrata ranged from none to 5%, average 14.5%～32.6%. Contribution ranked highest to lowest is plasic pipes, hollow bricks, cement pipes, tiles and stones. Our results would be helpful in the choosing of more appropriate settlement substrata for cultured holothurians.
6. Growth, survival and food sources of sea cucumber(Apostichopus japonicus) co-cultured with scallop(Chlamys Farreri) in Ponds were determined using stable carbon isotope biomarker. The results showed the specific growth rate (SGRw) and survival rate(SR)of A japonicus were (1.43±0.72)% and (96.67±5.77)% that both were higher than monoculture holothurians, (1.29±0.62)%d-1 and (93.3±5.77)%, respectively. Contribution of food sources of monoculture sea cucumber ranked highest to lowest was microalgae, bacteria and sediment and their contributon was 62%,20.6% and 17.4%, while co-cultured ones were microalgae, bacteria, sediment, scallop biodeposits and their contributon was 51.9%,22%,15.8% and 10.3%. Results here would be of some help in the choosing of appropriate cocultued aquatic aquatic organism for holothurians.
1、为避免海泥用作刺参饲料添加物的负面作用,寻找其替代物是当前刺参规模化养殖需要解决问题之一。本实验拟采用稳定碳同位素比率法研究用黄泥代替海泥的可行性。结果：添加20%黄泥、海泥的绿藻粉分别按7.5%刺参幼参体重投喂60天,刺参的特定生长率SGR分别为(1.95±0.13)%d-1及(1.72±0.17)%d-1,黄泥、海泥对刺参食物来源贡献分别为(9.78±1.37)%、(9.89±1.47)%,绿藻粉对刺参食物来源贡献分别为(90.22±1.37)%、(90.11±1.47)%,以上各项指标在黄泥组刺参和海泥组间均无显著性差异(P>0.05)。结果表明,用黄泥代替海泥完全可行,并能降低养殖成本及对浅海底质环境的破坏。
2、通过分析刺参养殖池塘内采集到的样品的碳稳定同位素值(δ13C值)发现：底栖动物的稳定碳同位素值与其食物来源密切相关,故可用于研究其食物来源组成。刺参的主要食物来源按δ13C值范围大致可划分为3类：即水体中的植物性碳源(主要为底栖硅藻等)、动物性碳源(小型底栖生物等)以及沉积物有机质(底泥有机质SOM)。研究结果表明,稳定碳同位素是分析刺参养殖池塘食物网结构的有效工具。
3、实验采用室内实验与池塘实验相结合方法,池塘实验设计了5,10,15,25,35头/m2的刺参养殖密度,日投饵率5%刺参体重,投喂60d来研究人工饲料对幼刺参生长的贡献。结果：(1)人工饲料对室内水族箱、池塘围隔内刺参的最终体重影响显著(F=148.541,P=0.003;F=6.301,P=0.018),饲料组围隔内刺参的最终体重均高于同密度对照组;放养密度对围隔内刺参的最终体重影响极显著(F=13.009,P<0.01),随密度增大最终体重减小；(2)人工饲料对水族箱刺参、补充饲料与密度及其互作对池塘刺参的最终δ13C影响均显著(F=3.0,P=0.032;F=18.856,P<0.01;F=4.954,P=0.003; F=20.421,P<0.01)。投喂补充饲料的围隔内刺参的δ13C值的随着密度的增大由(-13.262±0.183)‰下降到(-15.102±0.189)‰。(3)补充饲料在不同的密度下对刺参的食物贡献差异均极显著(F=24.439,P<0.01),随着密度增大,人工饲料对刺参食物贡献增加,由最低密度5头/m2时(3.78±2.98)%的食物贡献,增加到最高密度35头/m2时的(29.48±3.31)%。本实验为刺参增养殖实践人工饲料的使用提供了科学的参考。
4、采用碳氮双稳定同位素法分析了池底改造的池塘养殖刺参的食物来源。实验结果表明：实验刺参的特定生长率SGR为1.9%d-1,其成活率为64.7%。应用IsoSource软件计算得出：不同食物来源对刺参生长贡献分别为细菌26.6%(13～34%)、沉积物21.5%(4～40%)、大型藻类19.1%(0～52%)、小型底栖生物18%(0～49%)及硅藻14.8%(0～41%)。可见,刺参为杂食的沉积物食性生物。
5、利用稳定碳同位素比率法定量分析了石头、瓦片、塑料管、空心砖和水泥管这五种附着基为养殖刺参提供的食物来源情况。结果发现,底栖微藻在五种附着基表层物中均是主要碳源(39.9%～73%),而沉积物和细菌分别占7.3%～34.5%与9.7%～29.6%。不同附着基表层物在刺参食物来源中放的贡献平均在14.5%～32.6%范围,由大到小依次为塑料管>空心砖>水泥管>瓦片>石块。本研究结果对刺参规模化养殖中附着基的选择有一定的借鉴作用。
6、利用稳定碳同位素法定量分析了混养栉孔扇贝(Chlamys (Azumapecten)Farreri)对刺参(Apostichopus japonicus)生长及食物来源的影响。结果发现：混养模式下刺参的特定生长率SGR为(1.43±0.72)%d-1、存活率为(96.67±5.77)%,两者均略高于单养模式的(1.29±0.62)%d-1和(93.3±5.77)%,上述指标均无显著性差异(P=0.709；P=0.519)。在单养刺参的食物来源,由贡献率大到小分别为微藻62%、细菌20.6%、沉积物17.4%；在混养刺参的食物来源,按贡献率由大到小分别为微藻51.9%、细菌22%、沉积物15.8%、扇贝沉降物10.3%。本研究结果对刺参混养模式的选择有一定的借鉴作用。
1. How to eliminate negative effects of sea mud in feed for juvenile sea cucumber (Apostichopus japonicus) is an unsolved problem. Here we propose a method that using stable carbon isotope ratio to prove possibilities of replacement for sea mud with yellow soil. The results showed that feeding juveniles with algae contained 20% yellow soil and sea mud at 7.5% their body weights and for 60 days, the SGR of A. japonicus were (1.95±0.13)%, (1.72±0.17)%, contributions of yellow soil and sea mud to food sources of juveniles were (9.78±1.37)% and (9.89±1.47)%, contributions of green algae to food sources of juveniles were(90.22±1.37)% and (90.11±1.47)%, there was no significant difference for the same index between treatments of sea mud with yellow soil(P>0.05). These results support replacing sea mud with yellow soil is of great feasibility and can save costs and minimize the disturbance of Shallow sea beds.
2. By analysing stable carbon isotopic values (δ13C values) of the organisms collected in a sea cucumber farming pond, it is found that the 813C values of one animal has a close relation with that of the digested food and may be useful in elucidating their food sources. Using the stable carbon isotopic tracers may help confirm the importances and compositions of food source for filter feeders and predators. The investigations show that the main food sources of the Apostichopus japonicus can be divided into three groups in terms of stable carbon isotopic composition:benthic diatoms, meiofauna and organic matter in sediments (POM and SOM).The results lead to a conclusion that theδ13C value is useful tracer to investigate the food web structure of coastal ponds.
3. Stable carbon isotope ratios (δ13C) were used to estimate the contribution of supplemental feed to growth of juvenile sea cucumber cultured in pond enclosures. Five different culture densities (5,10,15,25 and 35 individuals /m2) of sea cucumbers were cultured in pond enclosures for 60 days, keeping constant feeding ration of 5% initial weight. The results were as followed:1) supplemental feed had significant effects on final weights of sea cucumbers both in aquarium and enclosure study(F=148.541, P=0.003; F=6.301, P=0.018, final weights sea cucumbers fed on supplemental feed were higher than that in unfed enclosures, significant differences were observed among different densities (F=13.009, P<0.01), a decrease in final weight with increasing stocking density was described.2) Supplemental feed had significant effects onδ13C values of sea cucumbers in aquariums, Supplemental feed, density had significant effects onδ13C values of sea cucumbers in aquariums, Supplemental feed, and density had significant interaction onδ13C values of sea cucumbers in aquariums. Theδ13C values of sea cucumbers fed on supplemental feed showed a descending trend, ranged from (-13.262±0.183)‰to (-15.102±0.189)‰, as the density increased from 5 to 35 inds./m2. It was calculated that a higher contribution of supplemental feed to sea cucumber growth was favored when density was increased, reaching a minimum (3.78±2.98)% at 5 inds./m2 and a maximum (29.48±3.31)% under the highest stocking condition.The contributions of supplemental feed showed significant differences among different densities (F=24.439, P<0.01). Food limitation may account for a fact of lower final weights while higher feed contributions were obtained under high density conditions.Results from this work should be an aid to developing intensive culture and optimum feed for holothurians.
4. Food sources of sea cucumber cultured in a bottom-renovated pond were determined using stable carbon and nitrogen isotope biomarkers. Based on computing result of the IsoSource software, Apostichopus japonicus's diet consisted of bacteria(13%～34%), sediments(4%～40%), diatoms(0～41%), macroalgae(0～52%), and meiofauna(0%～49%). Playing a primary role in the feeding of sea cucumber, bacteria contributed average 26.6% possible food sources, sort by size, contributions of sediments, macroalgae, meiofauna and diatom were 21.5%,19.1%,18%,14.8%, respectively.
5. Food contribution of stones, tiles, plasic pipes, hollow bricks and cement pipes to cultured sea cucumber (Apostichopus japonicus a) in enclosures were determined using stable carbon isotope biomarker.The surface organic matter of five settlement substrata consisted of diatoms, sediments, bacteria and bacteria. Playing a primary role in the surface organic matter, diatom contributed verage 39.9%～73% possible food sources, sort by size, contribution of sediments and bacteria were 17.3%～34.5% and 9.7%～29.6%, respectively. Contributions of the surface organic matter of five settlement substrata ranged from none to 5%, average 14.5%～32.6%. Contribution ranked highest to lowest is plasic pipes, hollow bricks, cement pipes, tiles and stones. Our results would be helpful in the choosing of more appropriate settlement substrata for cultured holothurians.
6. Growth, survival and food sources of sea cucumber(Apostichopus japonicus) co-cultured with scallop(Chlamys Farreri) in Ponds were determined using stable carbon isotope biomarker. The results showed the specific growth rate (SGRw) and survival rate(SR)of A japonicus were (1.43±0.72)% and (96.67±5.77)% that both were higher than monoculture holothurians, (1.29±0.62)%d-1 and (93.3±5.77)%, respectively. Contribution of food sources of monoculture sea cucumber ranked highest to lowest was microalgae, bacteria and sediment and their contributon was 62%,20.6% and 17.4%, while co-cultured ones were microalgae, bacteria, sediment, scallop biodeposits and their contributon was 51.9%,22%,15.8% and 10.3%. Results here would be of some help in the choosing of appropriate cocultued aquatic aquatic organism for holothurians.
引文
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