Immune Response of the Ridgetail White Prawn Exopalaemon carinicauda After Exposure to the Dinoflagellate Prorocentrum minimum

The dinoflagellate Prorocentrum minimum is known to affect the normal physiological function of Exopalaemon carinicauda by inducing oxidative stress,apoptosis,and cellular injury.To study the effects of P.minimum on the immune defense system of shrimp,E.carinicauda were exposed to 5×10^(3)cells mL^(-1)and 5×10^(4)cells mL^(-1)of P.minimum for 336 h in treatment groups,while E.carinicauda cultured in filtered seawater was employed as control.The total hemocyte counts(THC),hemocyanin concentration(HEM),and the activity of alkaline phosphatase(AKP)in hemolymph serum,as well as expressions of six immunity-related genes in hemocytes,hepatopancreases and gills were determined.The exposure of P.minimum significantly reduced the THC,HEM concentration and AKP activity in hemolymph serum.Immunity-related genes expressed differently in hemocytes,hepatopancreases and gills.Compared with the control group,the expressions of Crustin and pro PO in hemocytes were significantly up-regulated in the treatment groups,while the up-regulated expressions of LGBP,Lysozyme and Serpin were only found in the group exposed to 5×10^(4)cells mL^(-1)of P.minimum.In the gills of E.carinicauda exposed to P.minimum,the down-regulation of ALF,proPO and Serpin,up-regulation of LGBP and Lysozyme,as well as unaffected Crustin were observed.In hepatopancreases,the up-regulated expressions of LGBP,Crustin,Lysozyme,Serpin and proPO(only in 5×10^(3)cells mL^(-1)of P.minimum group)were found in the treatment groups.After exposure to P.minimum for 336 h,shrimps were injected with Vibrio parahaemolyticus and WSSV.The results showed that the mortality rates of shrimp in the treatment groups were significantly increased with a dose-dependent effect,which suggests that exposure to P.minimum may reduce the immunity of E.carinicauda.The research indicates that hemocytes and hepatopancreases play important roles in protecting the shrimp immune response to harmful algae,while the protection effect of hemolymph serum and gills may be suppressed.Since the exposure to P.minimum depressed the immunity of E.carinicauda,further studies are needed to confirm whether the presence of the algae will affect the susceptibility of shrimp to pathogens.

Effects of Salinity Fluctuation and Dietary Traditional Chinese Herbal Medicines on Survival,Growth and Immunity of Exopalaemon carinicauda

[Objective]The aim of this study was to provide a theoretical basis for the large-scale breeding of Exopalaemon carinicauda. [Method]Effects of salinity fluctuation （S0,S3,S6 and S9 represented salinity fluctuations of 0,3,6 and 9 respectivley） and dietary traditional Chinese herbal medicines （0,1%,2% and 4% respectively） on survival,growth and immunity of E.carinicauda were studied,and its experiment lasted for 60 d. [Result]The results showed that specific growth rate,feed conversion efficiency and superoxide dismutase activity of E.carinicauda in the treatment group with 1% dietary traditional Chinese herbal medicine and S6 salinity fluctuation were higher than those in other treatments,which was significantly higher than the treatment groups with S0,S3 and S9 salinity fluctuation at the same adding level of 1% dietary traditional Chinese herbal medicine. The regression analysis indicated that at S0,S3 and S6 salinity fluctuation,the optimal adding level of dietary traditional Chinese herbal medicines in feeds decreased with the increase of salinity fluctuation,accounting for 2.74%,1.77% and 0.51% respectively. At four adding levels of dietary traditional Chinese herbal medicines,the optimal salinity fluctuation of E.carinicauda also decreased with the increase of dietary traditional Chinese herbal medicines in feeds,accounting for 5.26,4.95,3.54 and 1.97 respectively. [Conclusion]Salinity,dietary traditional Chinese herbal medicines and their interactions have a significant effect on survival,growth and immunity of E.carinicauda,which calls for attentions in the breeding.

池塘养殖脊尾白虾(Exopalaemon carinicauda)感染血卵涡鞭虫的研究

通过现场取样、显微镜检、细菌培养、组织病理、电镜观察以及PCR检测等研究手段,对2008年9月浙江省舟山市普陀区某梭子蟹养殖塘混养的脊尾白虾"白浊病"进行了系统的病原学研究。结果表明,病虾体色发白,反应迟钝,血淋巴液呈浊白色,严重患病白虾血淋巴中的血细胞几乎没有或者极少,代之以大量与血细胞形似的寄生虫;组织病理切片及电镜观察发现,病虾心脏、肝胰腺、肌肉、鳃部等组织寄生有大量单细胞寄生虫,该寄生虫虫体形态特征、显微结构及宿主临床症状与本实验室之前研究报道的感染三疣梭子蟹和锯缘青蟹的血卵涡鞭虫类似;采用已建立的针对血卵涡鞭虫ITS1基因的引物对"白浊病"病虾的血淋巴样品进行PCR检测,可扩增出585bp左右的特异条带。综合以上研究结果,确定该寄生虫为血卵涡鞭虫(Hematodinium sp.)。有关脊尾白虾感染该寄生虫的报道在国内外尚属首次。

急性氨氮胁迫对脊尾白虾(Exopalaemon carinicauda)抗氧化系统酶活力及GPx基因表达的影响

为了研究氨氮毒性作用对脊尾白虾(Exopalaemon carinicauda)抗氧化及脂质过氧化作用的影响,将脊尾白虾暴露于低浓度氨氮(4.79±0.12)mg·L-1、中浓度氨氮(9.05±0.18)mg·L-1、中高浓度氨氮(17.64±0.25)mg·L-1和高浓度氨氮(34.87±0.46)mg·L-1中72h,于胁迫后1、3、6、12、24、48、72 h测定血淋巴和肝胰腺中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)、丙二醛(MDA)含量以及GPx基因相对表达量,结果显示:除低浓度氨氮组外,氨氮胁迫6～24 h,脊尾白虾血淋巴和肝胰腺组织SOD、CAT活力都有不同程度的升高,胁迫48～72 h上述指标则均被抑制;氨氮胁迫过程中血淋巴GPx活力和GPx基因表达显著高于对照组(P<0.05),而肝胰腺中GPx基因表达在48～72 h有所下降;血淋巴和肝胰腺中MDA含量随着胁迫时间的延长呈现上升的趋势,且上述指标在肝胰腺中的活力较血淋巴高。实验结果表明,氨氮胁迫3～24 h对脊尾白虾SOD和CAT活力具有一定的诱导作用,但胁迫48～72 h则抑制其活性;脂质过氧化产物MDA含量在整个胁迫过程中有增加的趋势;肝胰腺可能是脊尾白虾主要的氨氮代谢中心。

南极磷虾(Euphausia superba)与脊尾白虾(Exopalaemon carinicauda)营养学特征分析及鲜味评价

为科学评价南极磷虾这一新兴资源,同时为小型虾的高值化加工利用提供参考,选取体型上相近的南极磷虾与脊尾白虾作为研究对象,通过测定基本营养成分、氨基酸组成和脂肪酸组成分析2种虾的营养学特征,通过测定游离氨基酸与呈味核苷酸,进而计算滋味活性值与味精当量(equivalent umami concentration,EUC)评价其鲜度。结果表明,南极磷虾和脊尾白虾的粗蛋白质量分数具有显著性差异(P<0.05),分别为13.25%和16.41%,粗脂肪质量分数同样差异显著(P<0.05),分别为3.12%和1.10%。2种虾的脂肪酸组成有明显不同,南极磷虾的不饱和脂肪酸含量相对更高。南极磷虾的水解氨基酸总量低于脊尾白虾(P<0.05),但必需氨基酸所占比例更高,更加符合FAO/WHO推荐的蛋白质营养的理想模式。南极磷虾和脊尾白虾均是次黄嘌呤核苷酸和游离谷氨酸对鲜味起主要贡献作用,其EUC分别为11.01 g MSG/100 g和12.98 g MSG/100 g。南极磷虾与脊尾白虾都具有较高的营养价值,且具备理想的鲜味特征,在普通食品、调味品开发方面具有广阔的市场前景。

脊尾白虾(Exopalaemon carinicauda)体长和体重遗传力的估计

构建了脊尾白虾(Exopalaemon carinicauda)49个全同胞家系,包括28个半同胞家系,分别测定了30日龄和50日龄个体的体长和体重。利用SPSS软件的一般线性模型(GLM)过程,计算表型变量的方差组分,估计体长和体重性状的遗传力。结果表明,脊尾白虾30日龄和50日龄体长的遗传力估计值分别在0.14—0.35和0.07—0.31之间,脊尾白虾30日龄和50日龄体重的遗传力估计值分别在0.12—0.23和0.14—0.33之间;雄性遗传方差组分均显著大于雌性遗传方差组分,说明雄性遗传方差组分存在显著的父本效应。经过t检验,父系半同胞、母系半同胞方差组分估计的遗传力均未达到显著水平,全同胞方差组分估计的遗传力达到极显著水平。因此可以认为基于全同胞方差组分估计的遗传力是对脊尾白虾两个生长阶段体长和体重狭义遗传力的无偏估计值。本研究结果表明脊尾白虾体长和体重性状属于中度遗传力,选择育种对于脊尾白虾早期生长的改良具有较大潜力。

干露胁迫对脊尾白虾(Exopalaemon carinicauda)HSP70和ferritin基因表达的影响

采用常规急性实验方法,研究了不同干露胁迫条件对脊尾白虾(Exopalaemon carinicauda)成活率的影响。结果表明,保持虾体湿润并用冰块降温的P组干露胁迫12h后成活率为75%,显著高于其它各实验组(P<0.05)。采用实时荧光定量PCR方法研究了不同干露胁迫条件对脊尾白虾血细胞和肝胰腺组织中热休克蛋白70(HSP70)和ferritin基因表达的影响。干露胁迫能诱导脊尾白虾血细胞、肝胰腺HSP70基因的表达上调,肝胰腺中的高表达时间相对血细胞中出现的较早;干露胁迫能诱导湿润低温P组脊尾白虾血细胞和肝胰腺ferritin基因的表达上调,并显著高于对照组(P<0.05),而且各组织ferritin基因表达的上调时间具有差异性,血细胞最先上调。其余实验组脊尾白虾各组织ferritin基因表达均下调,并显著低于对照组(P<0.05)。上述结果表明,在脊尾白虾受干露胁迫的耐受范围内,HSP70和ferritin基因发挥抗氧化功能。

脊尾白虾(Exopalaemon carinicauda)3个野生群体mtDNA 16S rRNA序列差异及长臂虾科系统进化关系

采用PCR扩增和序列测定等技术,对脊尾白虾3个野生群体共计58个个体的线粒体16SrRNA基因片段进行初步研究。结果表明,在获得的长度为509bp核苷酸序列中,共检测到7个变异位点、8种单倍型,除象山湾群体外其它两个群体遗传多样性水平都较低。AMOVA分析表明,象山湾与莱州湾群体有显著的遗传差异,其余群体间的遗传差异不显著。另外,将本研究所得序列与GenBank中长臂虾科11个种的16SrRNA基因片段序列进行比较后,发现其聚类关系与传统分类略有不同,并依据16SrRNA序列变异百分比推测了长臂虾科12个种的大致分化时间。

脊尾白虾(Exopalaemon carinicauda)ferritin基因克隆及表达分析

采用RT-PCR及RACE技术获得了总长度为755bp的脊尾白虾ferritin基因cDNA序列。该基因序列5’和3’的非编码区分别为112bp和146bp,开放阅读框497bp,推测编码169个氨基酸,预测蛋白分子量19.1kDa,等电点5.46,该基因命名为Ecfer基因。利用Neighbor-joining法构建系统进化树分析结果表明,Ecfer基因氨基酸序列与无脊椎动物中的甲壳动物聚为一支(同源性为64%—67%)。此外,Ecfer基因氨基酸序列与脊椎动物铁蛋白H链同源性为53%—57%。采用荧光定量RT-PCR研究Ecfer基因在组织中以及经WSSV刺激后血淋巴、肝胰腺中的表达变化情况。研究结果表明,Ecfer在肝胰腺、卵巢、肌肉、鳃和血淋巴中均有分布。注射WSSV后3h和6h,肝胰腺和血淋巴Ecfer基因表达较对照组均显著上调(P<0.05),并具有显著的时间差异性。说明Ecfer基因可能参与了脊尾白虾抵抗WSSV入侵的免疫反应。

脊尾白虾(Exopalaemon carinicauda)不同地理群体的生化遗传分析

采用聚丙烯酰胺不连续凝胶垂直电泳方法,进行了中国北方沿海的河北渤海湾、山东莱州湾、山东胶州湾和江苏海州湾4个脊尾白虾(Exopalaemon carinicauda)地理群体间的生化遗传差异分析。结果表明,12种同工酶共有16个基因座位,其中Ldh-1、Adh-1、Sdh-1、s-Mdh-1、Gcdh-1和Me-1共6个座位表现为多态;平均每个座位等位基因有效值Ae=1.22911.3009,平均预期杂合度He=0.13300.1675,平均观察杂合度Ho=0.17970.2474,遗传距离D=0.00500.0345;聚类分析表明,渤海湾群体和莱州湾群体亲缘关系最近,其次为胶州湾群体与海州湾群体。本研究结果表明,脊尾白虾遗传多样性处于较高水平,种质资源状况良好,有利于其种质资源的保护开发及遗传改良。

脊尾白虾(Exopalaemon carinicauda)维甲酸X受体基因克隆及其在温盐胁迫和蜕皮周期中的表达分析

为了明确脊尾白虾维甲酸X受体基因在环境胁迫和蜕皮周期中的作用,本研究通过RACE技术克隆了脊尾白虾(Exopalaemon carinicauda)维甲酸X受体c DNA全长,命名为Ec RXR基因,该c DNA序列全长为1323 bp,开放阅读框(ORF)为855 bp,编码一个284个氨基酸组成的蛋白质,分子量为30.918 k Da,理论等电点为PI为6.788;Ec RXR基因推导氨基酸序列经Blastp在线比对分析显示,Ec RXR与已知甲壳动物RXR的同源性为71%—90%;系统进化树分析结果显示,脊尾白虾Ec RXR的氨基酸序列与日本沼虾RXR的氨基酸序列聚为一支。荧光定量RT-PCR分析表明,Ec RXR基因在各组织中均有表达,而在眼柄相对表达量较高,血淋巴中最低。Ec RXR基因在蜕皮周期中的表达规律表明,Ec RXR基因表达在不同蜕皮时期存在明显变化,在眼柄、肝胰腺、胃和肠中整体呈现上升趋势,在鳃中呈现先下降后上升的趋势,在表皮中呈现逐渐降低的趋势。Ec RXR基因在温度、盐度胁迫过程中的表达规律分析结果发现,温度、盐度胁迫均可显著改变Ec RXR基因在鳃和肝胰腺中的表达模式,温度胁迫下Ec RXR基因在鳃中呈先上升后下降的表达趋势,肝胰腺中整体呈先上升后下降再上升再下降的表达趋势;盐度对鳃和肝胰腺的调控模式相同,均呈先升高后下降的变化趋势。本研究结果表明Ec RXR基因在脊尾白虾蜕皮发育、酶活性调控及渗透压调节中发挥重要作用,为深入研究甲壳动物维甲酸X受体基因的功能提供了重要的基础信息。

脊尾白虾(Exopalaemon carinicauda)组织蛋白酶D基因的克隆及其表达分析

采用RACE技术获得了总长为1853bp的脊尾白虾(Exopalaemon carinicauda)组织蛋白酶D基因全长cDNA序列。该基因5′和3′非编码区分别为120bp和572bp,开放阅读框为1161bp,推测编码386个氨基酸,预测分子量为42.36kDa,理论等电点为7.54,命名为EcCatD基因。同源性和系统进化分析表明,EcCatD基因与斑节对虾、美洲螯龙虾的相似性分别为86%和80%,与斑节对虾和美洲螯龙虾紧密聚为一支。荧光定量RT-PCR结果表明,EcCatD基因在血细胞中的相对表达量最高,其次为肝胰腺。该基因在鳗弧菌和WSSV感染后的脊尾白虾血细胞和肝胰腺中的表达量显著增加,并具有不同的时空表达趋势。结果表明EcCatD基因在脊尾白虾免疫反应中具有重要作用。

脊尾白虾(Exopalaemon carinicauda)α2-巨球蛋白cDNA全长的克隆和表达分析

根据本实验室前期获得的脊尾白虾(Exopalaemon carinicauda)α2-巨球蛋白基因EST序列,采用c DNA末端快速扩增(Rapid amplification of c DNA end,RACE)技术克隆获得脊尾白虾α2-巨球蛋白基因c DNA全长,命名为Ecα2M基因。该基因全长4823 bp,由4413 bp的开放阅读框、64 bp的5'端非编码区以及346 bp的3'端非编码区组成。开放阅读框编码1470个氨基酸,分子量为163.0 k Da,理论等电点为5.03。序列分析显示,Ecα2M序列N端含有23个氨基酸组成的信号肽。同源性分析显示,脊尾白虾Ecα2M氨基酸序列与罗氏沼虾(Macrobrachium rosenbergii)α2M的同源性最高,达到80%。荧光定量PCR分析结果显示,Ecα2M基因在血细胞、肝胰腺、肌肉、鳃、卵巢、眼柄、胃及肠中均有表达,其中在血细胞中的相对表达量最高。感染鳗弧菌和WSSV后,脊尾白虾血细胞中Ecα2M的相对表达量于6 h达到最大值且显著高于对照组(P<0.05),肝胰腺中Ecα2M的相对表达量于3 h达到最大值且显著高于对照组(P<0.05),相对表达量变化具有明显的时间差异性。

氟苯尼考对脊尾白虾(Exopalaemon carinicauda)免疫和抗氧化功能的影响

为研究氟苯尼考对脊尾白虾(Exopalaemon carinicauda)免疫和抗氧化功能的影响,以低、中、高(10、20和40 mg/kg·BW)3种剂量氟苯尼考药饵连续投喂脊尾白虾5 d,对照组投喂基础饲料。于停止投喂药饵后的第3、6、12、24、48、96、168和240 h取血淋巴,测定血蓝蛋白(HEM)含量、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)和总抗氧化能力(T-AOC)等指标的变化情况。结果显示,低、中剂量组HEM含量均在3 h时间点显著高于对照组(P<0.05),高剂量组在3–48 h显著高于对照组(P<0.05);低、中剂量组ACP活性在3 h和6 h显著高于对照组(P<0.05),高剂量组在3–168 h显著低于其他3组(P<0.05);低、中剂量组AKP活性在3 h均显著高于对照组(P<0.05),6–48 h显著低于对照组(P<0.05),高剂量组在3–48 h显著低于对照组(P<0.05);低、中剂量组T-SOD活性整体高于对照组,分别在6 h和24 h达到最高值(P<0.05),高剂量组在24 h和48 h显著低于对照组(P<0.05);低、中、高剂量组CAT活性均在3 h和6 h显著高于对照组(P<0.05),之后在12–96 h显著低于对照组(P<0.05),于24 h达到最低值(P<0.05);低、中、高剂量组T-AOC活性均在3–96 h显著低于对照组(P<0.05),于48 h达到最低值(P<0.05),且出现剂量效应。本研究结果为氟苯尼考在海水养殖生产中使用的安全评估提供了数据支持。

致病性粪肠球菌对脊尾白虾(Exopalaemon carinicauda)免疫相关基因表达的影响

为了研究致病性粪肠球菌(Enterococcus faecalis)对脊尾白虾(Exopalaemon carinicauda)免疫相关基因表达的影响,分别对健康脊尾白虾注射生理盐水和粪肠球菌,测定了不同时间点血细胞和肝胰腺中酚氧化酶原(proPO)、C型凝集素(CTL)、超氧化物歧化酶(SOD)、组织蛋白酶D(Cat D)和组织蛋白酶L(Cat L)基因的相对表达量变化。结果显示,与对照组相比,感染粪肠球菌后,血细胞和肝胰腺中pro PO基因表达量均于6 h达到最大值(P<0.05),随后逐渐降低至对照组水平;血细胞和肝胰腺中CTL基因表达量分别于12 h和6 h上升至最大值,随后逐渐降低;血细胞中CTL基因表达量于72 h虽有降低,但仍显著高于对照组(P<0.05);血细胞和肝胰腺中SOD基因表达量分别于6 h和3 h达到最大值,随后逐渐降低,并于72 h达到正常水平;血细胞和肝胰腺中Cat D基因表达量变化趋势基本一致,呈现先降低后升高、然后显著降低再升高的趋势;血细胞和肝胰腺中Cat L基因表达量分别于6 h和3 h显著降低,随后逐渐升高至最大值,并于72 h达到对照组水平。研究表明,长时间的粪肠球菌感染对脊尾白虾免疫相关基因表达影响显著,本研究所用免疫相关基因均可作为粪肠球菌感染疾病的监测指标。

Cloning of catalase and expression patterns of catalase and selenium-dependent glutathione peroxidase from Exopalaemon carinicauda in response to low salinity stress

Catalase （CAT） and selenium-dependent glutathione peroxidase （Se-GPx） play a vital role in protecting organisms against various oxidative stresses by eliminating H202, The objective of this paper is to evaluate the roles of these antioxidant molecules in the ridgetail white prawn Exopalaemon carinicauda in response to low salinity stress. A complementary DNA （cDNA） containing the complete coding sequence of CAT was cloned from the hepatopancreas using reverse-transcription polymerase chain reaction （RT-PCR） and rapid amplification of cDNA ends. The full-length cDNA of CAT （2 649 bp） contains a 5＇-untranslated region （UTR） of 78 bp, a 3＇- UTR of 1 017 bp, with a poly （A） tail, and an open reading frame of 1 554 bp encoding a 517-amino-acid polypeptide with predicted molecular mass of 58.46 kDa and estimated isoelectric point of 6.64. This CAT sequence contained the proximal active site signature （60FDRERIPERWHAKGAG76）, proximal heme-ligand signature sequence （350RLFSYPDTH358） and three catalytic amino acid residues （His71, Asn144 and Tyr354）. Sequence comparison showed that the CAT deduced amino acid sequence of E. carinicauda shared 68%-92% of identities with those of other species. Quantitative real-time PCR analysis revealed that CAT mRNA was widely expressed in the hepatopancreas （highest）, hemocyte, eyestalk, heart, gill, muscle, ovary and stomach. Under low salinity stress, CAT and GPx mRNA expression levels both in the gill and hepatopancreas increased significantly at the first 48 h and 6 h respectively, indicating a tissue- and time-dependent antioxidant response in E. carinicauda. All these results indicate that E. carinicauda CAT is a member of the CAT family and might be involved in the acute response against low salinity stress.

Effect of salinity on growth and first sexual maturity of Exopalaemon carinicauda(Holthuis, 1950)

To determine the optimal salinity for growth and first sexual maturity of ExopaIaemon carinicauda, the effects of salinity on growth and reproductive performance of early juvenile prawns of E. carinicauda were evaluated under laboratory conditions. Postlarvae from the same female broodstock were reared at six salinity levels （5, 10, 15, 20, 25, and 30） for 12 weeks. The specific growth rate （SGR） and survival rate （SR） under different salinity levels in the first 6 weeks were calculated and compared. SGR was significantly influenced by salinity. Prawns reared in salinity of 10 grew significantly faster （P〈0.05） than those reared in salinities of 5, 20, 25, and 30. However, ANOVA confirmed that there was no significant effect among the six salinity levels on SR. For the next 6 weeks, the body length （BL）, body weight （BW） at the first sexual maturity, and the age at median sexual maturity （As0） of females were measured and compared. Female prawns reared at salinity of 10 presented significantly shorter A50, but no significant differences among the six salinity levels for BL and BW at the first sexual maturity were observed. Based on the above information, the optimal salinity for growth and first sexual maturity of juvenile E. carinicauda is approximately 10.

Antioxidant Response of Ridgetail White Prawn Exopalaemon carinicauda to Harmful Dinoflagellate Prorocentrum minimum Exposure and Its Histological Change

The dinoflagellate Prorocentrum minimum,one of the most widespread red tide causing species,affects marine aquaculture and ecosystems worldwide.In this study,ridgetail white prawn Exopalaemon carinicauda were exposed to P.minimum cells(5 × 104 cells mL-1) to investigate its harmful effects on the shrimp.Antioxidant activities and histological changes were used as indicators of health status of the shrimp.In 72 hours,the mortality of E.carinicauda was not affected,but its antioxidant response and histology were statistically different from those of control.Elevated superoxide dismutase(SOD) and glutathione peroxidase(GPX) activities and depressed catalase(CAT) activity were observed in gill;while increased SOD,glutathione S-transferase(GST),CAT activities and modulated GPX activity were observed in hepatopancreas.Thus,antioxidant activities in gill and hepatopancreas seem to respond differentially to harmful alga exposure.Increased malondialdehyde(MDA) content in early a few hours indicates the damage of the antioxidant defense system.Although MDA content recovered to a low level thereafter,a series of histological abnormalities including accumulation or infiltration of hemocytes,tissue lesions and necrosis were discovered in gill and hepatopancreas.Exposure to P.minimum induced sublethal effects on E.carinicauda,including temporary oxidative damage and histological injury.

Exposure to Prorocentrum minimum Induces Oxidative Stress and Apoptosis in the Ridgetail White Prawn, Exopalaemon carinicauda

Prorocentrum minimum is a bloom-forming, planktonic mixotrophic dinoflagellate, and can cause stress in shrimp ponds. In this study, healthy Exopalaemon carinicauda were exposed to 5 × 10^4 cells mL^-1 P. minimum for 72 hours to investigate the adverse effect of P. minimum on shrimps. Elevated superoxide dismutase(SOD) activity and malondialdehyde(MDA) content, reduced total antioxidant capacity(T-AOC) and catalase(CAT) activity, and regulatory glutathione peroxidase(GPX) activity were found in the hemolymph of E. carinicauda after exposure to P. minimum. In this study, P. minimum exposure induced oxidative stress and caused significant oxidative damage to E. carinicauda. P. minimum exposure increased the expression of HSP70 gene in the hemocyte, gills and hepatopancreas. Compared with the enhanced level of caspase-3 gene mR NA in the hemocyte and gills, the up-regulation of caspase-3 gene in the hepatopancreas was only observed from 3 to 6 h, and then the mRNA level of glutathione-S-transferase(GST) gene increased. These results indicated that GST might be involved in the shrimp hepatopancreas’ defense against P. minimum exposure. The present study demonstrates that exposure to P. minimum could induce oxidative stress and apoptosis in E. carinicauda. The SOD activity, HSP70 and GST(in the hepatopancreas) were evoked to protect cells from oxidative stress and apoptosis. This study will provide new insights into the toxic mechanism of P. minimum on shrimps.

Inhibition of Breast Cancer Resistance Protein(BCRP) by Ko143 Can Affect Pharmacokinetics of Enrofloxacin in Exopalaemon carinicauda

Adenosine triphosphate-binding cassette transporter breast cancer resistance protein(BCRP) exists highly in the apical membranes of epithelia, and is involved in drug availability. Ko143 is a typical inhibitor of BCRP in rodents. The synthetic antibacterial agent enrofloxacin(ENRO) is a fluoroquinolone employed as veterinary and aquatic medicine, and also a substrate for BCRP. BCRP gene highly expressed in the hepatopancreas and intestine of Exopalaemon carinicauda as was determined with real-time quantitative reverse transcription-polymerase chain reaction(RT-q PCR) method. The effects of Ko143 on the abundance of BCRP m RNA and ENRO pharmacokinetics in E. carinicauda were studied. The m RNA abundance of BCRP decreased significantly in hepatopancreas and intestine(P < 0.05) after Ko143 treatment. Co-administration of Ko143 significantly changed the pharmacokinetics of orally administered enrofloxacin, which was supported by higher distribution half-life(t_(1/2α)), elimination half-life(t_(1/2β)), area under the curve up to the last measurable concentration(AUC_(0-t)), peak concentration(C_(max)) and lower clearance(CL/F). These findings revealed that Ko143 downregulates BCRP expression in hepatopancreas and intestine, thus affects the pharmacokinetics of orally administered enrofloxacin in E. carinicauda. The drug-drug interaction can be caused by the change in BCRP activity if ENRO is used in combination with other drugs in shrimp.