Distribution of Bacteria Injected in Body of Giant Black Shrimp,Penaeus Monodon

Distribution of injected Vibrio anguillarum in body of P,,naeus monodon was studied with immunohistochemical method. Bacteria could be detected throughout the experiment in some individuals; however in lymphoid tissue, gill, heart and haemolymph of all vibrio injected shrimp, the bacteria could be observed only 5 min after injection. The bacteria density in haemolymph, haemolymph of the hepatopancreas and gills decreased with time. In the lymphoid organ and heart, the bacteria density was the highest 48 h after injection, then decreased. Nodules could be formed in the heart, lymphoid organ and injection site.     

注入丁醇调节重非水液相密度的微空隙试验模拟

密度大于水的重非水液相(DNAPLs)有机污染物在重力作用下向地下介质深部迁移从而增加污染范围。前人通过一维砂柱和二维砂箱试验发现利用密度调节技术可降低DNAPLs向下迁移的风险,但目前缺乏微观尺度上密度调节影响DNAPLs迁移的定量观测。本研究试验模拟丁醇注入微空隙调节四氯乙烯(PCE)的密度,通过建立非水相中染色PCE浓度、密度与灰度的定量关系,监测注入丁醇后空隙介质中非水相密度的动态变化,基于空隙中代表性非水相PCE受力情况分析其运移状态,揭示空隙尺度介质性质和密度调节程度对DNAPLs迁移的影响。试验结果表明：丁醇注入后,PCE浓度和密度迅速下降,离散状PCE与丁醇有效接触面积大且起效快;当非水相密度降至略大于水相密度时,非水相受毛细力和摩擦力的影响停止向下迁移;当非水相密度小于水相密度时,非水相才在注入压力与浮力的作用下克服毛细力、重力和摩擦力向上迁移;注入压力、摩擦力、毛细力、浮力与重力影响着空隙中非水相的迁移行为,空隙半径越大,毛细力对调节PCE向上迁移的影响越小;密度比水小的丁醇注入介质后向上迁移,因此丁醇从DNAPLs下端注入可提高修复效率。试验证实了向空隙介质中注入...     

哈维氏弧菌GST核酸疫苗制备及其对斜带石斑鱼的免疫保护作用

通过同源引物从致病性哈维氏弧菌(Vibrio harveyi)ZJ0603基因组中克隆GST的开放阅读框(ORF),构建真核表达质粒pcDNA-GST。大量抽提重组质粒后,于背鳍基部肌肉注射重组质粒免疫斜带石斑鱼(Epinephelus coioides),分析重组质粒的免疫效果。通过核酸水平检测重组质粒在鱼体肝、肌肉、头肾和脾脏组织的分布;用ELISA法检测鱼体血清的抗体水平,用Western-blot检测目的蛋白的表达情况。结果表明:该序列全长615 bp;免疫7 d后,鱼体中均有质粒分布;斜带石斑鱼血清中产生抗GST的高效抗体(1∶4 096);相应的目的蛋白也在鱼体中成功表达。攻毒后,疫苗免疫保护率达80%,表明GST可作为防治哈维氏弧菌病有效候选抗原。     

Hematological Changes m White Spot Syndrome Virus-Infected Shrimp, Fenneropenaeus chinensis (Osbeck)

The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifesta- tions of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and dis- integrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular bemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.     

Effects of Hypoxia on Dopamine Concentration and the Immune Response of White Shrimp （Litopenaeus vannamei）

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolvmph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5mgL-1 DO groups at 12h and 6h, and then returned to normal after 24h and 12h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0mgL-1 DO group became stable after 12h,while granular cells did so after 24h, The THC and different haemocyte count (DHC) in the 1.5mgL-1 DO group became stable alter 24h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5mgL-1 DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 It, respec-tively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activ-ity were obviously related to the exposure time under hypoxic conditions.     

Effects of hypoxia on dopamine concentration and the immune response of White Shrimp (Litopenaeus vannamei)

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolymph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5 mg L⁻¹ DO groups at 12 h and 6 h, and then returned to normal after 24 h and 12 h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0 mg L⁻¹ DO group became stable after 12 h, while granular cells did so after 24 h. The THC and different haemocyte count (DHC) in the 1.5 mg L⁻¹ DO group became stable after 24 h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5 mg L⁻¹ DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 h, respectively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activity were obviously related to the exposure time under hypoxic conditions.     

Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.     

DYNAMIC STUDIES ON FLAGELLAR REGENERATION IN DUNALIELLA SALINA

INTRODUCTIONMicroalgalbiotechnologyhasattractedmoreandmoretheinterestofscientistsandpeopleex ploitingitscommercialpossibilitiesasitisthebiotechnologythatcanmatchbacterialbiotechnologyinthecomingcenturyintheaspectofeconomicreturnoninvestment.Dunaliellasalinaisoneofthemostdetailedlystudiedunicellulargreenalgae,hassomeuniquescientificvalues,andgreatestpotentialformasscultureasafoodsource ,andisrichestalgalsourceofglycerolandβ carotene(Borowitaka,1 988;Ginzburg,1 987) .Optimizingperformanceo…     

Dynamic studies on flagellar regeneration inDunaliella salina

Flagella, the basic locomotive organ in algae, as well as bacteria and some cells of animals or high plants, would be damaged in the well stirred mass culture due to the strong forces caused by the fast mixing impellers. The dynamic regeneration of the flagella in deflagellatedDunaliella salina was studied microscopically by using a bench-top flat-bottom photobioreactor. The results showed that 90 minutes was necessary for the repair of flagella, after which half of the cells became motile as their flagella generated within 120 minutes and nearly all of the cells could swim freely within 180 minutes. Contribution No. 3503 from the Institute of Oceanology, Chinese Academy of Sciences.     

注射黄芪多糖对吉富罗非鱼c型溶菌酶基因表达量的影响

将黄芪多糖(APS)用无菌生理盐水配制成2 mg/mL和20 mg/mL针剂,腹腔注射吉富罗非鱼,以注射无菌生理盐水为对照。24 h后分别提取吉富罗非鱼鳃、头肾、肝脏、脾脏等组织中的总RNA并反转录成cDNA,利用Real-time PCR方法对不同组织中基因表达进行定量分析。结果表明:吉富罗非鱼腹腔注射20 mg/mL高剂量APS后,其鳃、头肾、肝脏等三个组织中的Lysozyme-c基因表达量显著高于对照组(P<0.05);注射2 mg/mL低剂量APS后,Lysozyme-c基因表达量仅在脾脏中出现显著上调(P<0.05)。APS可通过诱导Lysozyme-c基因在鳃、头肾、肝脏和脾脏等组织在的表达量,来提高吉富罗非鱼的机体免疫力。     

溶藻弧菌对凡纳滨对虾血清中一氧化氮及氧自由基的影响

对凡纳滨对虾注射浓度为1×105、1×106、1×107mL-1的溶藻弧菌悬液,在注射后4、12、24、48、72和120 h于腹血窦取血,测定其血清中一氧化氮(NO)、丙二醛(MDA)含量,以及一氧化氮合酶(NOS)、超氧化物岐化酶(SOD)活力。结果表明,注射溶藻弧菌后,凡纳滨对虾血清中NO含量及NOS活力显著高于对照组,且NOS活力最大值出现时间较NO早;而SOD活力先升高后下降,MDA含量在SOD活力开始下降后逐渐升高,说明一氧化氮系统对凡纳滨对虾感染的溶藻弧菌有清除作用,并能在一定程度上对损伤过程中生成的氧自由基产生作用。     

10种CpG-ODNs对罗非鱼抗海豚链球菌感染的作用

采用人工合成的10种未甲基化CpG-ODNs序列分别注射健康的吉富罗非鱼Oreochromis niloticus,48 h后注射海豚链球菌(Streptococcus iniae)进行人工感染,分别于感染24 h和48 h后检测鱼体血清溶菌酶(Lysozyme,LSZ)、超氧化物歧化酶(Superoxide dismutase,SOD)和碱性磷酸酶(Alkaline phosphatase,AKP)的活性,并记录死亡率。结果显示:注射ODN-205的罗非鱼LSZ、AKP和SOD活性变化均不显著(P>0.05),推测注射ODN-205后罗非鱼能较好的维持机体的平衡,增强机体的抗菌能力;注射ODN-1681、1670、2133和2006的罗非鱼LSZ活性逐渐降低,而注射ODN-1826、2143、1668、2102、1669的罗非鱼LSZ活性呈现先升高后降低的趋势。AKP活性的变化趋势同LSZ活性变化大体相似,而SOD活性前后变化不显著(p值>0.05)。注射CpG-ODNs的实验组抑制海豚链球菌感染的作用差异显著(p值<0.05),其中以注射ODN-205组的死亡率最低(18%),其次为注射ODN-1670、1681、2006和2133组,死亡率均低于30%,而ODN-1669组和生理盐水组的死亡率均超过90%。实验表明,注射一定量的ODN-205、1670、1681、2006和2133能明显提高吉富罗非鱼抗海豚链球菌感染能力。     

脂多糖、苯酚、硫酸铜对红笛鲷非特异性细胞毒性细胞受体基因表达的影响

应用Real-time PCR技术,研究脂多糖(lipopolysaccharide,LPS)、苯酚、硫酸铜刺激红笛鲷(Lutjanussanguineus)后非特异性细胞毒性细胞受体(NCCRP-1)基因在不同组织里的表达差异。结果发现,LPS刺激红笛鲷24 h后NCCRP-1在红笛鲷头肾、脾脏、胸腺、肝脏、心脏、脑、肌肉和肠组织中均有表达,其中头肾表达量最高,脾脏次之,然后依次是肝脏、脑、肌肉、胸腺和肠,心脏表达量最少。LPS、苯酚和CuSO4刺激红笛鲷后,随着刺激时间的增长,NCCRP-1表达量在各组织达到峰值的时间不同。以头肾为模式组织,RT-PCR的结果显示,红笛鲷NCCRP-1在LPS、苯酚和CuSO4的刺激下的表达模式相似,随着时间的增加NCCRP-1表达量逐渐增加,分别在24、9、12 h处达到最高,达到对照组的52、30、24倍左右,之后表达量开始下降。免疫组织化学表明,NCCRP-1只在头肾、脾脏和胸腺的特定细胞中表达。     

硝化细菌对罗非鱼苗培育环境及抗病力的影响

于罗非鱼鱼苗培育水体中引入不同浓度的硝化细菌,检测主要水质因子,测定罗非鱼与抗病力有关的酶的活力,研究微生态调控对水质和对罗非鱼抗病力的影响。结果显示,硝化细菌浓度为100/L时,氨氮含量相对于对照组降低25.05%,亚硝酸氮含量降低45.16%,COD值降低12.33%,差异显著(P<0.05);鱼苗培育成活率相对于对照组高7.58%,体长增加22.18%,体重增加46.15%;幼鱼的抗菌酶、碱性磷酸酶(AKP)和超氧化物歧化酶(SOD)活力相对于对照组分别提高18.60%、59.19%和49.89%,差异显著(P<0.05),组织过氧化物酶(POD)活力差异不显著。表明不同浓度的硝化细菌可显著改善罗非鱼苗培育环境的水质,增强罗非鱼苗的抗病力。     

LHRH-A和hCG对南方鲶血清促性腺激素水平的影响

运用放射免疫分析法测定了促黄体释放激素类似物 (LHRH A)和人绒毛膜促性腺激素(hCG)对南方鲶血清促性腺激素 (GTH)水平的影响 ,结果表明 :LHRH A和hCG都能提高南方鲶一龄幼鱼血清GTH水平 ,hCG与LHRH A结合注射后血清GTH水平显著高于单独注射hCG或LHRH A后的血清GTH水平。LHRH A和hCG结合注射可诱导性成熟南方鲶排卵 ,同时排卵过程中伴随着血清GTH水平的急剧上升 ,在排卵后迅速下降 ;注射同样剂量的药物而未排卵的血清GTH水平虽有升高 ,但升高的幅度不及排卵鱼。这种血清GTH水平的急剧增高对诱导南方鲶卵母细胞最后成熟和排卵具有重要作用。     

Application of Lactic Acid Bacteria (LAB) in Freshness Keeping of Tilapia Fillets as Sashimi

Aquatic products are extremely perishable food commodities. Developing methods to keep the freshness of fish represents a major task of the fishery processing industry. Application of Lactic Acid Bacteria(LAB) as food preservative is a novel approach. In the present study, the possibility of using lactic acid bacteria in freshness keeping of tilapia fillets as sashimi was examined. Fish fillets were dipped in Lactobacillus plantarum 1.19(obtained from China General Microbiological Culture Collection Center) suspension as LAB-treated group. Changes in K-value, APC, sensory properties and microbial flora were analyzed. Results showed that LAB treatment slowed the increase of K-value and APC in the earlier storage, and caused a smooth decrease in sensory score. Gram-negative bacteria dominated during refrigerated storage, with Pseudomonas and Aeromonas being relatively abundant. Lactobacillus plantarum 1.19 had no obvious inhibitory effect against these Gram-negatives. However, Lactobacillus plantarum 1.19 changed the composition of Gram-positive bacteria. No Micrococcus were detected and the proportion of Staphylococcus decreased in the spoiled LAB-treated samples. The period that tilapia fillets could be used as sashimi material extended from 24 h to 48 h after LAB treatment. The potential of using LAB in sashimi processing was confirmed.     

Immune response of Pseudosciaena crocea to the injection of Vibrio alginolyticus

For the investigation of anti-infection immune response of Pseudosciaena crocea, 160 healthy fish samples were categorized into infected and control groups. Each individual fish in the infected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension of Vibrio alginolyticus in density of 2×107 CFU/ml, while each individual in the control group was injected i.p. with 0.2 ml sterile saline solution (0.85%). It was observed that the artificial injection of V. alginolyticus significantly inc...     

Ontogeny of the immune system in rock bream Oplegnathus fasciatus

Histogenesis of the immune system and specific activity of superoxide dismutase(SOD) were studied in rock bream Oplegnathus fasciatus from fertilization to 50 days after hatching(DAH).The pronephric tubule primordium developed in the embryo,14 h 30 min post fertilization.The spleen anlage was observed between the swim bladder and the intestine at 5 DAH,and the thymus was formed as a paired structure under the pharyngeal epithelium above the gill arch at 10 DAH.The order of the immune organs becoming lymphoid was the pronephric kidney(10 DAH),thymus(15 DAH) and spleen(21 DAH).As the embryo developed,the specific activity of SOD gradually increased until hatching,but subsequently SOD activity continuously decreased to a minimum at 14 DAH.After the spleen became lymphoid,the specific activity of SOD was relatively stable.It is suggested that the immaturity of the lymphoid organs and low specific activity of SOD was the cause of the high mortality of fingerlings 12 to 16 DAH.     

Drying of Micro-Encapsulated Lactic Acid Bacteria—Effects of Trehalose and Immobilization on Cell Survival and Release Properties

Lactic acid bacteria (LAB) were encapsulated with alginate, gelatin and trehalose additives by the extrusion method and dried at 4 ℃. The microcapsules were generally spherical and had a wrinkled surface with a size of 1.7 mm ± 0.2 mm. Trehalose as a carbohydrate source in the culture medium could reduce acid production and performed no function in the positive proliferation of LAB. Using trehalose as a carbohydrate source and protective medium simultaneously had a benefit in the protection of LAB cells dur...     

过热蒸汽对熟制小龙虾优势腐败菌的杀菌动力学及其机理

【目的】研究过热蒸汽杀菌对熟制小龙虾优势腐败菌的杀菌效果。【方法】从腐败熟制小龙虾中分离腐败菌,根据16S rDNA基因序列信息鉴定其种属,研究过热蒸汽温度、流量和杀菌时间等的处理参数对小龙虾优势腐败菌的杀菌效果,并利用Weibull模型拟合其杀菌动力学过程。通过研究过热蒸汽处理对优势腐败菌菌悬液的电导率和紫外吸收的影响,以及通过扫描电子显微镜对菌体的微观结构进行观察,初步探讨其杀菌机理。【结果】熟制小龙虾优势腐败菌为短小芽孢杆菌(Bacillus pumilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、蜡样芽孢杆菌(Bacillus cereus)和赖氨酸芽孢杆菌(Lysinibacillus macroides)。在过热蒸汽杀菌初期阶段,优势腐败菌的残活率迅速下降,随着杀菌时间延长,残活率下降速率变得缓慢,最后趋于稳定,说明过热蒸汽能完全杀灭小龙虾优势腐败菌。Weibull模型能较好拟合过热蒸汽对4种优势腐败菌的杀菌动力学,其R2≥0.935。Weibull模型得到的预测值与实测值接近程度较高。通过对模型的分析,提高过热蒸汽温度和增大流量,更容易到达灭活腐败菌的效果。过热蒸汽杀菌后,菌悬液的电导率值和吸光度迅速上升,之后趋于平缓;通过对微观结构的观察,发现杀菌后腐败菌的细胞膜完整性遭到严重破坏。【结论】过热蒸汽杀菌技术应用于小龙虾产品加工具有一定的可行性。