运用RAPD技术对笛鲷属3种鱼的群体遗传学分析

应用RAPD技术，从9个系列180种随机引物中筛选出16种，对笛鲷属的画眉笛鲷(Lutja-nus vitta）、金焰笛鲷(Lutjanus fulviflamma)_，金带笛鲷(Lutjanus vaigiensis)进行种群内及种群间遗传学分析，并利用UPGMA确定了它们之间的亲缘关系。结果表明，三种鱼的种内遗传多样性指数(H)分别为，画眉笛鲷0．1949，金带笛鲷0．1107，金焰笛鲷0．1673；三者的多态位点比例(P)分别为69．4％、47．8％和59．0％；金带笛鲷具有较低的遗传多样性，且金带笛鲷与金焰笛鲷有较近的亲缘关系。三种鱼共检出10个可作为种的特异性鉴定的条带，可用于种质鉴定。     

恩诺沙星在红笛鲷体内的药代动力学

在水温26±2℃、盐度28条件下,恩诺沙星(enrofloxacin,EF)按5 mg/kg的剂量腹腔注射和口灌红笛鲷(Lutjanus sanguineus),用反相高效液相色谱法研究恩诺沙星在红笛鲷体内的药代动力学和组织分布。结果显示:注射和口灌后红笛鲷血浆的药时数据均符合一级吸收二室模型;血药达峰时间(tp)分别为0.75和1.5 h,血药质量浓度峰值(cmax)分别为5.22和3.94μg/mL,药时曲线下面积(AUC)分别36.98和17.24 mg/(L.h),消除半衰期(t1/2β)分别为13.28和9.18 h;恩诺沙星在组织中分布较广,注射和灌服给药肌肉、肝脏和肾脏的cmax分别为3.53和3.40μg/g、4.08和3.98μg/g、11.23和5.40μg/g,tp分别为1.5和2.0 h、1.0和2.0 h、0.75和1.5 h,AUC分别33.17和23.12 mg/(L.h)、39.36和18.20 mg/(L.h)、98.97和56.69 mg/(L.h);EF在红笛鲷体内消除速度较慢,注射和灌服给药肌肉、肝脏和肾脏的t1/2β分别为22.08和83.32 h、41.52和94.47 h、20.94和21.81 h。表明恩诺沙星注射给药在红笛鲷体内的吸收和消除均快于口灌给药。     

凡纳滨对虾池塘沉积物中氮、磷形态的赋存特征

【目的】研究凡纳滨对虾(Litopenaeusvannamei)养殖池塘沉积物中氮、磷形态的赋存特征。【方法】于对虾养殖周期的前、中和后期对池塘沉积物采样,测定和分析沉积物中各形态氮和磷含量。【结果与结论】对虾养殖池塘沉积物中总氮质量分数(1131.36±349.00)mg/kg,有机氮平均占总氮的74.14%,养殖中期总氮及有机态氮含量相对较高。无机氮主要为固定态铵,平均占75.71%,可交换态氮主要由硝态氮和氨态氮组成。总磷质量分数(854.17±254)mg/kg,主要磷形态为铁结合态磷(Fe-P)、自生钙磷(ACa-P)和闭蓄态磷(Oc-P),养殖周期内沉积物中Fe-P含量变化频繁,较活跃。垂直方向上,总氮及有机氮在次表层4～6 cm含量较高,总磷、Fe-P及Oc-P均有表层富集现象。对虾养殖过程中池塘沉积物中的氮磷总量未见明显累积,生物活性磷(BAP)在总磷中比例增加,池塘沉积物内源磷负荷加重。氮和磷分别主要以有机态氮和生物活性磷形式存在,养殖过程使沉积物中内源磷释放潜力增加。     

凡纳滨对虾高位池养殖系统的水质理化状况

对凡纳滨对虾高位养殖池水质环境状况进行了研究。结果表明,养殖期间虾池水体透明度前期较高,中后期较低;虾池pH变化在7.89~9.02之间,变化幅度较小。虾池水体悬浮物数量和CODMn随养殖时间延长而持续升高,变化范围分别为18.5~162.3 mg/L和3.52~14.58 mg/L。虾池水体中营养盐各月份波动较大,无机氮的数量变化在0.206~1.621 mg/L之间,含量逐渐升高,磷酸盐数量变化在0.009~0.067 mg/L之间,含量逐渐下降。水环境中N、P比较高,平均为62.7。养殖水体中后期处于严重富营养化状态。     

南海海域4种笛鲷的微卫星DNA分析

采用聚丙烯酰胺凝胶电泳技术,从20对西大西洋笛鲷(Lutjanus campechanus)的微卫星引物中筛选适用于红鳍笛鲷(L.erythopterus)、勒氏笛鲷(L.russellii)、紫红笛鲷(L.argentimaculatus)和约氏笛鲷(L.johnii)基因组分析的微卫星引物,分析4种笛鲷的遗传多样性及系统发生。经反应条件的优化,从20对引物中筛选出17对可稳定扩增出特异片段的引物。通过测序证实扩增产物含微卫星位点后,以该17对引物对4个种的群体进行遗传多样性分析。其中16对可在约氏笛鲷、勒氏笛鲷、紫红笛鲷基因组中扩增出重复性好的特异性条带,分别有65%、65%、60%呈现出种内多态;15对可在红鳍笛鲷中扩增出重复性好的特异性条带,并且全部呈现出种内多态。四种笛鲷中,红鳍笛鲷的多态性最高,高度多态基因座占检测座位的66.67%;勒氏笛鲷的多态性最低,低度多态基因座占43.75%。获得3个种间特异性分子标记。用DS和DA两种方法计算4种笛鲷的遗传距离,构建了系统发生树。     

两种体色画眉笛雕的遗传多样性及分子标记

根据体侧纵带颜色差异,暂将画眉笛鲷(Lutjanus vitta)分为黄带画眉笛鲷和褐带画眉笛鲷,应用RAPD技术,选用22个引物对黄带画眉笛鲷与褐带画眉笛鲷进行遗传多样性及分子标记研究。结果表明,黄带画眉笛鲷与褐带画眉笛鲷的遗传距离为0.832 9,其多态位点比例(P)分别为66.99%和85.28%,遗传多样性指数(H)分别为0.161 7和0.289 3,种内遗传距离(D)分别为0.243 5和0.431 9,表明褐带画眉笛鲷的遗传多样性比黄带画眉笛鲷丰富。共得到OPP14-629bp和OPP14-674bp等11个特异性分子标记,可用于两种笛鲷的鉴定。     

红鳍笛鲷和紫红笛鲷种类和群体的矢耳石地标点法识别

【目的】探讨矢耳石地标法在笛鲷种内及种间中的判别作用。【方法】利用2017年购自广西北海、海南文昌、广东阳江的87尾红鳍笛鲷(Lutjanus erythropterus)和76尾紫红笛鲷(Lutjanus argentimaculatus)成鱼的矢耳石样本,基于地标点法分析耳石的形态差异,运用判别分析检验耳石形态差异在2种笛鲷的种间和同种不同群体间的判别功效。【结果】位于听沟前中部交叉点的两个地标点10、11贡献较大,解释了耳石形态变异的64.88%～65.85%,表明两种笛鲷耳石形态的种间差异和种内群体差异主要集中于听沟前中部。基于耳石形态的地标点方法对2种笛鲷的种间判别成功率为97.4%和100.0%;红鳍笛鲷和紫红笛鲷的种内不同群体判别成功率分别为85.7%、83.3%、80.0%和94.1%、78.1%、81.5%。【结论】矢耳石地标点法可作为2种笛鲷种间和种内判别的有效工具。     

徐闻珊瑚礁保护区营养盐时空分布特征

2006年8月(夏季)、2006年11月(秋季)、2007年2月(冬季)和2007年4月(春季)在徐闻珊瑚礁自然保护区灯楼角至流沙湾近岸海域调查徐闻珊瑚礁保护区的营养盐变化及空间分布特征。结果表明:徐闻珊瑚礁保护区水域溶解态无机氮以NO3--N为主,其含量超过总溶解无机氮的50%;各站点NO2--N含量相对较低,冬季NO3--N和NH4+-N含量均高于其它季节;无机磷含量在0mg/L～0.030mg/L之间变化;活性硅含量表现为夏秋季节高、冬春季节低;表层水体硝酸盐氮/无机磷原子比值(N/P)夏季较低,不存在无机磷受限情况,而冬季N/P整体较高,此时水体主要受无机磷限制。     

红笛鲷IRAK-1基因克隆及哈维弧菌感染后的组织表达

用同源克隆和RACE的方法克隆红笛鲷(Lutjanus sanguineus)白细胞介素1受体相关激酶1(Interleukin-1receptor-associated kinase 1,IRAK-1)基因的cDNA全序列,并用荧光定量PCR分析其在健康鱼及哈维弧菌感染后红笛鲷的组织表达。结果表明,该序列全长3 207 bp(登录号KF728204),包含5′非编码区(UTR)202 bp,3′UTR752 bp,开放阅读框(ORF)2 253 bp,编码750个氨基酸。根据推导的氨基酸序列预测其蛋白分子质量为82.6 ku,理论等电点为6.59。IRAK-1包含1个N端死亡结构域、proST结构域、中央激酶结构域和C末端结构域。荧光定量PCR分析显示,红笛鲷IRKA-1基因在肝脏和皮肤的表达量最高,其次是心脏、鳃、肌肉和胸腺,其余组织的表达量较低。哈维弧菌侵染红笛鲷后,各组织中IRAK-1基因mRNA表达量均呈上调趋势,肝组织中变化最显著。     

红笛鲷弧菌病血清流行病学研究方法的建立

以红笛鲷Lutjanus sanguineus弧菌病的主要病原菌——溶藻弧菌Vibrio alginolyticus为抗原制备兔抗血清,以辣根过氧化酶标记的羊抗兔血清为酶标二抗,建立了酶联免疫吸附试验检测技术。通过棋盘滴定法测定,得出溶藻弧菌菌悬液的最佳工作浓度为5×108mL-1,兔抗溶藻弧菌血清的最佳稀释度为1∶16000,酶标羊抗兔抗体的最佳稀释度为1∶2000。应用本实验建立的间接双抗体夹心法对红笛鲷进行现场检测,患病红笛鲷特异性抗体的检出率为70%,外观健康红笛鲷的检出率为20%。结果表明,本实验所建立的酶联免疫吸附试验(ELISA)检测法可以用于红笛鲷弧菌病的血清流行病学研究,而且可以检测出隐性感染状态下的红笛鲷。     

Assessment of nitrogen maintenance requirement in juvenile Black Porgy Acanthopagrus schlegeli

Nitrogen balance method and nitrogen-free diet were used in this study to determine nitrogen maintenance requirement (NM) and nitrogen maintenance requirement per unit metabolism body weight (NM’) of black porgy Acanthopagrus schlegeli. Fish with body weight (BW) of 50, 80, 120, 160 and 200 g were fed by the diets containing three graded levels of crude protein (380, 420 and 460 g/kg). The results from nitrogen balance experiment showed that the amount of nitrogen deposition varied from 0.15 to 0.31 mg/g BW...     

Assessment of nitrogen maintenance requirement in juvenile Black Porgy <Emphasis Type="Italic">Acanthopagrus schlegeli</Emphasis>

Nitrogen balance method and nitrogen-free diet were used in this study to determine nitrogen maintenance requirement (NM) and nitrogen maintenance requirement per unit metabolism body weight (NM′) of black porgy Acanthopagrus schlegeli. Fish with body weight (BW) of 50, 80, 120, 160 and 200 g were fed by the diets containing three graded levels of crude protein (380, 420 and 460 g/kg). The results from nitrogen balance experiment showed that the amount of nitrogen deposition varied from 0.15 to 0.31 mg/g BW per day, accounting for 12.2% to 21.1% of nitrogen intake. The amount of fecal nitrogen excretion varied from 0.21 to 0.32 mg/g BW per day, accounting for 16.3% to 21.6% of nitrogen intake. The endogenous nitrogen excretion, a main part of nitrogen consumption varied from 0.79 to 0.97 mg/g BW per day, accounting for 63.3 % to 68.0% of nitrogen intake. Positive correlation was found between NM and body weight, while a negative correlation was found between NM of unit body weight and the growth duration. No significant differences (p>0.05) were found in NM′ among different growth stages. The average of NM′ was 0.485 7mg/g per day. The results from nitrogen-free diet experiment showed that a negative correlation between NM and feed intake of nitrogen-free diet. NM increased with the decrease of feed intake of fish. The average of NM was 0.482 9 mg/g BW per day that was close to 0.483 8 mg/g BW obtained from fish with 120 g BW in nitrogen balance experiment. The nitrogen balance method is recommended to be a better method for determining NM in consideration of fish stress and result stability. This study also provides a calculated result of the protein content in diets, which is necessary for maintaining fish body protein at different growth stages. The calculation was based on the amount of nitrogen required for maintaining body protein per kg BW. Supported by Scientific Research Project Grant (No.2004C100059) from the city government of Ningbo, China.     

Forms and Balance of Nitrogen and Phosphorus in Cage Culture Waters in Guangdong Province, China

In order to approach the characteristics of nitrogen and phosphorus pollution caused by cage culture and the balance of nitrogen and phosphorus during the process of cage culture, a monitoring was conducted in Daya Bay of Guangdong Province, China from April 2002 to Jane 2003. The results show that the concentrations of total nitrogen (TN) in the waters at the sites with five and ten years of cage culture history are 1.8 and 2.3 times of that at control site respectively. Ammonium (NH3-N) is the main form of nitrogen in spring while nitrate ( NO3 -) in winter. The concentrations of TN, total dissolved nitrogen (TDN) and dissolved organic nitrogen (DON) are highest in autumn. The concentration of phosphorus increases with the increasing of the culturing time, among which phosphate ( PO 34-) increases most obviously. The concentrations of total phosphorus (TP) and total dissolved phosphorus (TDP) are highest in autumn. The nitrogen and phosphorus are accumulated significantly in the sediment of cage culture area. The model of N balance in the cage culture area: bait (70.62%) fry (0.28%) input by tide (14.8%) release from sediment (14.3%) = harvest of adult fish (12.07%) deposition into sediment (28.75%) output by tide (56.18%) others (3.00%). The model of P balance: bait (83.11%) fry (0.17%) input by tide (12.23%) release from sediment (4.49%) = harvest of adult fish (8.43%) deposition into sediment (48.59%) output by tide (41.94%) others (1.04%). In one fish growth year, the contents of nitrogen and phosphorus in harvest of adult fish are only 17.0% and 10.1% of the contents of nitrogen and phosphorus in fish bait and fry, wherein 83% of nitrogen and more than 89% of phosphorus in fish bait became marine pollutants.     

Effects of dietary levels of carbohydrate,lipid, phosphorus and zinc on the growth,feed conversion and protein efficiency ratio of Nile tilapia (Oreochromis niloticus)

A 54-day feeding experiment was conducted on juvenile Nile tilapia using isonitrogenous, isocaloric semipurified diets. The carbohydrate content in the diet was 9%, 32% and 50%; the corresponding lipid content was 22.2%, 12%, and 4%. The diets were supplemented with 0.85% or 1.5% phosphorus and 40 mg/kg or 100 mg/kg zinc. The experiment was carried out in flow-through aquaria using dechlorinated tap water at 23–26°C. The experiment showed that the increase of the carbohydrate content in the diets resulted in a 43–249% increase in weight gain, a 27–59% decrease in feed conversion ratio, and a 65–121% increase in protein efficiency ratio. In fish fed diets containing 36–50% carbohydrate, an increase in supplemented phosphorus to 1.5% greatly increased the weight gain. On the contrary, a high content of supplemented zinc (100 mg/kg) inhibited growth and increased feed conversion ratio.     

福州市农田土壤养分空间变异特征

研究农田土壤养分空间分布特征及变异规律,分析地形因子对土壤养分空间分布的影响,对土壤养分的有效利用与管理具有重要意义。本文采用GIS和地统计学相结合的方法,揭示东南丘陵区福州市农田土壤养分（有机质、碱解氮、有效磷和速效钾）的空间分布特征和变异规律,并进一步探讨土壤养分含量与地形因子（地形起伏度、高程、地形湿度指数、沉积运输指数和坡度）之间的相关性。结果表明：福州市农田有机质、碱解氮、有效磷和速效钾含量范围分别为1.10~89.5 g/kg、 1.00~461 mg/kg、0.300~298 mg/kg、和4.00~399 mg/kg,其变异系数范围为35.3%~99.0%,均属于中等变异水平;区域内耕地土壤养分存在明显的空间丰缺差异,大部分地区有机质和有效磷含量较为丰富,碱解氮含量处于中等偏上水平,速效钾含量相对较低;有机质、碱解氮、有效磷和速效钾的块金系数分别为32.0%、37.3%、50.0%和50.0%,均呈中等强度空间自相关,表明同时受到结构和随机因素控制;有机质和碱解氮空间自相关尺度较大,且在步长小于0.3 km时,各方向（0°、45°、90°和135°）变化平稳,为各向同性,而有效磷和速效钾变程较小,且各方向变化较复杂,为各向异性。这些结果表明,政府需加强施肥指导,保持氮肥施用量的同时,合理增加钾肥,适当降低有机肥和磷肥施用。此外,在后续调查采样时,样点布设要考虑密度和方向性,适当加密有效磷和速效钾的采样,而有机质和碱解氮采样可以在此基础上适当减少样点。     

大亚湾网箱养殖区异养细菌和弧菌的数量动态

研究了大亚湾网箱养殖水体中细菌的数量动态,以探讨其变化规律与鱼病的关系。结果表明:总细菌、弧菌的数量变化各异,网箱内、网箱外和对照海区细菌数量分布与变化不同。养殖区异养细菌丰值期在4-5月份,5月份最高;网箱水体弧菌的丰值期在6-11月,最高在8月;网箱外丰值期在7-9月,最高在8月,对照海区最高值在9月,其他时间变化不大。无论是异养菌还是弧菌,细菌值大小顺序为网箱内、网箱外、对照海区。弧菌数量与部分环境因子相关性较为密切,但相关程度随不同空间而不同,可能受其他环境条件影响。弧菌的高值期与弧菌病的流行期相吻合,鱼体受伤或其他原因引起细菌感染也会使水体中细菌数量升高。     

Impacts of mariculture practices on the temporal distribution of macrobenthos in Sandu Bay,South China

Sandu Bay is located in the East China Sea and is characterized by high-density fish farming and kelp culture. Despite this, little is known about the impacts of these different mariculture practices on the local environment. We investigated the temporal variation in macrobenthos and environmental conditions at three sites in the bay (fish farming site, kelp culture site, and a control site). We collected water and sediment samples during nine cruises between May 2009 and February 2010. The density of macrobenthos peaked at the fish farming site in July (655 ind./m2) whereas density did not fluctuate as widely at the other two sites. Biomass varied significantly at both the control and kelp culture sites, but had only a single peak at the fish farming site in June (21.90 g/m2). The dominant species varied throughout the study period at the control and kelp culture sites, whereas a single terebellid species (Lysilla pacifica) dominated the macrobenthos at the fish farming site. The diversity index H’ increased at the control site beginning in February then decreased after May, whereas H’ was low at the other sites in December. The mean dissolved oxygen level was highest at the control site (6.59 mg/L) and lowest at the fish farming site (5.54 mg/L). DO levels were lowest at all sites in summer (July and August). The sediment acid volatile sulfide content was higher at the fish farming site (1.46 mg/g dry weight) than those at the kelp culture and control sites (1.22 and 0.14 mg/g, respectively). Our results suggest that mariculture practices have a clear impact on the benthic environment/ community in Sandu Bay.     

DYNAMICS OF LITTER DECOMPOSITION AND SEASONAL DYNAMICS OF PHOSPHORUS IN DECOMPOSED RESIDUA OF Calamagrotis augustifolia IN THE WETLAND OF THE SANJIANG PLAIN

Ｅｌｅｍｅｎｔｓｃｙｃｌｉｎｇａｎｄｅｎｅｒｇｙｆｌｏｗａｒｅｔｈｅｉｍｐｏｒｔａｎｔｒｅｓｅａｒｃｈｃｏｎｔｅｎｔｓｉｎｅｃｏｓｙｓｔｅｍｆｉｅｌｄｓａｎｄｒｅｓｉｄｕａｄｅ-ｃｏｍｐｏｓｉｔｉｏｎｉｓｔｈｅｋｅｙｐｒｏｃｅｓｓｏｆｎｕｔｒｉｍｅｎｔａｌｅｌｅｍｅｎｔｓｃｉｒｃｕｌａｔｉｏｎＳＷＩＦＴｅｔａｌ．１９７９．Ｔｈｅｒｅｓｉｄｕａｄｅ-ｃｏｍｐｏｓｉｔｉｏｎｒａｔｅｐｌａｙｓａｎｉｍｐｏｒｔａｎｔｒｏｌｅｉｎｄｅｃｉｄｉｎｇｔｈｅｐｒｏｄｕｃｔｉｖｉｔｙａｎｄｂｉｏｍａｓｓｏｆ…     

Dynamics of litter decomposition and seasonal dynamics of phosphorus in decomposed residua of Calamagrotis augustifolia in the wetland of the Sanjiang Plain

During the period of May to October in 1999, systematical studies were given to the rate of decomposition of Calamagrostis angustifolia litter, the phosphorus content and weight in decomposed residua of litter, and phosphorus content in the corresponding soil in the Sanjiang Plain. At the same time, the simulation models were listed in the paper. The results showed that the rate of weight lost of decomposition of Calamagrostis angustifolia litter is 29.80% and the maximum of daily rate of weight lost is 0.25%, which appeared in July. The change trend of phosphorus content and weight in the decomposed residua of litter is to reduce with the decomposing process, when it comes to the day of 157, the decrement amount of the both were respectively 57.69mg/kg and 1.6199mg, which were 72.80% and 76.30% of its previous amount. In addition, there is a polynomial minus correlationship of phosphorus content between the variation in corresponding soil and the decomposed residua of litter at the corresponding period. The study will be helpful to further understand the process and mechanism of biochemical cycling of nutrient elements in wetland ecosystems, in addition, it will also be helpful to the restoration and rebuilding of retrogressive wetlands and reasonable development and utilization of wetlands in the Sanjiang Plain. Foundation item: Under the auspices of the National Natural Science Fundation of China (No. 49771002), the Key B Items of Chinese Academy of Sciences (KZ951-B1-201-3) and the Innovative Project of the Chinese Academy of Sciences (KZCX2-302). Boigraphy: WANG Shi-yan (1974 -), male, a native of Jinan City, Shandong Province, Ph. D. candidate of Changchun Institute of Geography, the Chinese Academy of Sciences. His research interests include ecology of wetland environment and application of remote sensing and GIS.     

Effect of dietary phosphorus on the growth and body components of juvenile Synechogobius hasta

The effect of dietary phosphorus on the growth and body components of juvenile Synechogobius hasta was determined. Different percentages of dietary phosphorus (0.63, 0.77, 0.93, 1.06, 1.22 and 1.36) were tested by feeding the fish (body weight, 15.81 g ± 0.32 g; 20 individuals each group; 3 groups each percentage) at a surplus of 5%–10% above satiation for 35 d. Dietary phosphorus did not significantly affect the specific growth rate, feed intake, feed conversion ratio and protein efficiency rate. Nitrogen retention was found to be the highest in fish fed the diet containing 1.06% of phosphorus; however, this was not significantly different from other diets. Fish fed the diet containing 0.93% of phosphorus showed the highest phosphorus retention; similar phosphorus retention rates were found in fish fed the diets containing 0.77% and 1.06% of phosphorus. Fish fed the diet containing the highest percentage of dietary phosphorus were found to contain the least whole body lipid, lower than fish fed other diets (P<0.05). The protein content increased from 18.59% to 19.55% (although not significant) with the decrease of body lipid content (P>0.05). The contents of the whole body ash, whole body phosphorus and vertebrae phosphorus increased with dietary phosphorus percentage up to 1.06 (P<0.05), reaching a plateau after that. Dietary phosphorus did not significantly influence the muscle components (protein, lipid and moisture). Condition factor and hepatosomatic index were the highest in fish fed the diet containing 0.63% of dietary phosphorus; however, this was not significantly different from those of other diets. The second-order polynomial regression of phosphorus retention against dietary phosphorus identified a breakpoint at 0.88% of dietary phosphorus. However, the dietary requirement of phosphorus for maintaining maximum phosphorus storage determined by broken-line analysis of the contents of whole body phosphorus, and ash and vertebrae phosphorus was 1.06% of the diet.