Tilapia Head Glycolipid Alleviates Indomethacin-Induced Gastric Ulcer via Regulating Oxidative Stress and Inflammation Through COX/PGE2 Signaling Pathway in Adult Rats

The aim of this experiment was to investigate the ameliorative effect and molecular mechanism of tilapia head glycolipid(TH-GL)on indomethacin(IDM)-induced gastric ulcer in male Sprague Dawley(SD)rats.The gastric ulcer model was established by oral administration of 30mgkg^(-1) IDM after 7 days of TH-GL or omeprazole(OME)administration in rats.Then the macroscopic gastric injury symptoms,gastric mucosa protective factor cyclooxygenase 1(COX-1),cyclooxygenase 2(COX-2),prostaglandin E_(2)(PGE_(2)),the levels of oxidative stress,and inflammatory cytokine expression levels in the rats were analyzed.The experimental results showed that multiple ulcers appeared on the gastric surface of the rats in the model group.Compared to the model group,TH-GL significantly alleviated gastric ulcers and reduced the gastric damage index in rats.In addition,TH-GL significantly promoted the expression of constitutive enzyme COX-1 while inhibited the expression of inducible enzyme COX-2,and make PGE2 maintain at normal levels.TH-GL also inhibited oxidative stress and inflammatory responses,increased superoxide dismutase(SOD)activity and glutathione(GSH)content,decreased the level of malondialdehyde(MDA)and the content of pro-inflammatory factor.In conclusion,these results suggested that TH-GL could maintain the expression levels of COX-1 and PGE2 while inhibit the expression of COX-2 in the gastric of rat and then prevent IDM-induced gastric ulcer,which may be related to the regulation of oxidative stress and inflammatory response.Therefore,TH-GL might be a new option for the prevention of gastric diseases induced by IDM.

Effect of cooking processes on tilapia aroma and potential umami perception

Tilapia is a freshwater fish group with a sustainable prospect but suffers off-notes appearing during cooking processes.To promote pleasant odorants by thermal cooking processes,tilapia fillets were cooked in different ways(roasting,microwave-heating,boiling and steaming).Their aroma profiles were analysed with special focus on off-notes and umami-enhancing odorants by principal component analysis,and correlated with the heating time,colour,moisture and water activity by partial least squares regression analysis.Results showed that the“green”and“earthy”off-notes were highly correlated with the boiling process(excess of water,short heating time),while most of the umami-enhancing odorants had a high association with the roasting process(low water content,long heating time,better Maillard reaction).This study indicated that roasting is the most adapted cooking process promoting Maillard-derived aromas,umami-enhancing aromas and meanwhile,reducing off-notes.This research helps in understanding the off-note generation in tilapia and promoting desirable umami-enhancing odorants.

广东罗非鱼无乳链球菌分离鉴定及生物学特性分析

本研究旨在明确当前罗非鱼无乳链球菌流行状况和生物学特性,为鱼源无乳链球菌病防控提供新的数据。本研究于2019—2020年,从广东罗非鱼主养区茂名、湛江地区采集病样进行无乳链球菌的分离鉴定。用PCR方法对分离株血清型及4个主要毒力基因进行检测,通过纸片法对分离菌株进行31种抗菌药物药敏试验和多重耐药研究,利用斑马鱼模型进行了致病性试验。结果显示:从261份病样中分离到35株无乳链球菌;血清型检测表明无乳链球菌分离株的血清型均为Ia型;主要毒力基因的检测证明,cylE、sodA、gapC基因在所有鱼源分离株及人源、牛源和鱼源参考株中检出率均为100%,而scpB只在人源参考菌株中检出;药敏试验结果发现,无乳链球菌分离株对19种抗菌药敏感率为90%以上,对6种抗菌药耐药率为50%以上,分离株的耐药基因型和表型部分一致,无乳链球菌分离株均多重耐药,其中5重及以上耐药的菌株为27株,占总菌株数的77.1%;斑马鱼致病试验表明,无乳链球菌分离株可导致斑马鱼不同程度的发病死亡。本研究明确了当前罗非鱼源无乳链球菌的血清型、毒力基因、耐药特性和致病性,为广东乃至全国罗非鱼无乳链球菌病的防控提供了参考。

Water Quality and Growth Performance of Nile Tilapia Fries Fed on Insect-Based Feeds

Fish feed is one of the main constraints on the growth of aquaculture in Kenya. The lack of high-quality and competitively priced feeds means that the fish farmers are unable to expand their production. To ensure that aquaculture remains viable and sustainable, it is necessary to optimize production through the utilization of locally available feeding materials. Insect-based feeds are alternative least-cost feeding resource in fish production. Using alternative protein sources such as black soldier fly larvae (BSF) and daphnia meal in place of fishmeal (Omena and Dagaa) is an ingenious strategy for sustainable fish aquaculture. This study evaluated the effect of four treatments: BSF, daphnia, Omena from Lake Victoria and Dagaa from Lake Tanganyika on the fish pond environmental parameters and fish fries’ performance (body weight and length). The experiment was carried out in a greenhouse with 30 fingerlings in each experimental unit. The feeding trials took 23 days. Results indicated that the four treatments have a varied effect on the BW and L of fish fries. A high growth rate was observed in Daphia, followed by BSF, Omena and Dagaa in that order. On the other hand, the treatments have varied effects on pond parameters with BSF promoting too much growth of algae. BSF have higher amounts of ammonia, nitrates and phosphorus, which stimulate the high growth of algae.

外源物质对低盐罗非鱼糜蛋白构象及体外消化特性的影响

以微波超声波辅助制备的低盐罗非鱼糜凝胶为研究对象,分析谷氨酰胺转氨酶(TGase)、羟丙基二淀粉磷酸酯(HDP)、结冷胶和三者的复配物(THG)对其水分分布及蛋白质构象的影响;并通过体外模拟消化实验,探究不同外源物质对微波超声波辅助制备的低盐罗非鱼鱼糕体外消化特性的影响。结果表明:添加THG后,鱼糜结合水和不易流动水较空白组分别增加98.71%和14.75%,自由水含量显著减少(P<0.05);THG促进了α-螺旋向β-折叠、β-转角和无规则卷曲结构转化。与空白组相比,添加0.4%TGase和THG对低盐罗非鱼糕的胃排空速度、蛋白体外消化率及蛋白水解度均有正面作用;其中THG可显著促进蛋白质分解成粒径较小的聚集体(P<0.05),经胃-十二指肠消化结束后的消化产物颜色更加透明清晰,从激光共聚焦显微镜可观察到THG组的红色荧光亮点显著减少,反映其蛋白被消化得较为完全。由此可见,在THG的作用下,鱼糜中水分子与蛋白质结合更紧密,相对于单一的外源物质,THG更加明显地改变了蛋白质的构象,且更有利于疏水蛋白基团的暴露和蛋白间的相互作用,且对鱼糕的消化有促进作用。本研究的结果为罗非鱼糜凝胶特性研究及罗非鱼糕类食品的开发与应用提供理论参考。

尼罗罗非鱼无乳链球菌微滴式数字PCR检测方法的建立及临床应用

通过设计筛选引物和探针、优化反应浓度和退火温度,构建一种尼罗罗非鱼(Oreochromis niloticus)无乳链球菌(Streptococcus agalactiae)的微滴式数字PCR(dd PCR)检测方法,分析该方法的敏感性、特异性及重复性,并应用于临床样品检测。结果显示:当引物、探针浓度分别为0.9μmol·L^(-1)、0.3μmol·L^(-1)且退火温度为56.9℃时,建立的罗非鱼无乳链球菌dd PCR方法阴、阳性微滴分布界限明显,平均拷贝数高,有较高扩增反应效率;线性关系线良好(R^(2)=0.997 3),最低检测限为2.56 copies·μL^(-1);与猪链球菌2型、鱼类海豚链球菌和其他5种常见的水生动物疫病病原体无交叉反应;重复变异系数为3.15%;临床样品检测结果与实时荧光PCR方法结果的符合率100%,与细菌分离鉴定方法结果符合率为94.12%。结果表明,建立的罗非鱼无乳链球菌dd PCR检测方法灵敏度高、特异性强、重复性好,可对罗非鱼无乳链球菌感染的临床样品进行定量检测,为尼罗罗非鱼无乳链球菌的研究提供有益参考。

Impacts of Climate Change on Economic Performance of Tilapia Pond Operations in Minalin, Pampanga: A Case of Intensive Large-Scale Pond Culture

Pampanga is the top tilapia-producing province in Philippines. One of its municipalities, Minalin, is considered as one of its major centers of production. This study aims to determine the impacts of climate change hazards on the economic performance of tilapia-intensive pond culture in Minalin, Pampanga considering the threats to the industry, livelihoods, and food security. Results revealed that the economic performance of tilapia pond operations using semi-intensive large-scale culture was affected by climate change, as reflected in the reduction of yield among tilapia pond operators. This makes intensive tilapia pond farming in Minalin still a viable venture but will now require more than a year to pay for investment (variable and fixed) costs if under climate change risks. The study recommends strengthening capacity building related to climate-smart tilapia farming technologies for pond operators and to implement appropriate technologies to address climate change risks.

Effects of Garcinia kola “Bitter Kola” Powder on Sex Reversal, Survival and Growth Performance of Tilapia (Oreochromis niloticus L.)

This study was carried out in 2022 at the Itato agriculture community estate of fish hatchery in eastern Senegal with the aim of finding an alternative method to 17-α-methyl testosterone (MT) by using Garcinia kola “Bitter kola” powder to obtain Oreochromis niloticus male sex populations. 1800 fry of average size between 0.01 and 0.02 g were distributed in 12 aboveground tanks of 1 m3 each at a rate of 150 individuals/tank, following 6 duplicate treatments. The negative (B1 and B2) and positive (B3 and B4) control diets, to which between 0 and 60 mg/kg of 17alpha-methyl testosterone had been added respectively, were mixed with industrial feed containing 40% protein;the tank diets (B5 and B6), (B7 and B8), (B9 and B10) and (B11 and B12) contained 10 g, 20 g, 30 g and 40 g of Garcinia kola powder respectively for 1 kg of industrial feed con-taining 40% protein. These different treatments resulted in high proportions of males in the order of 97.03%;98.09% for the hormone and proportions of 94.03%;93.26% for 20 g of Garcinia kola extracts. In terms of weight growth, the best average weights for the hormone and Garcinia kola were 0.896 g and 0.980 g respectively. With regard to fish survival, the following results were recorded: 95.53% and 94.56% respectively for additions of 20 g and 30 g of Garcinia kola extracts to the feed supplied to the fish. On the other hand, a significant difference in the mortality rate of 35% and 5% respectively for the conventional hormone and 20 g of Garcinia kola extracts was observed. To conclude the study, it is recommended that further studies be carried out to determine the quality of the flesh, their life stages and the profile of the amino acids present in their proteins.

罗非鱼养殖尾水絮凝去除效果研究

罗非鱼养殖尾水中含有大量的残饲和粪便等颗粒悬浮物,这些物质是氮、磷营养元素的主要载体。将养殖尾水外排会导致周边水体富营养化。当前常见的尾水处理方式具有占地面积大、投资高、效率低等缺点,影响其推广使用。为解决这一问题,探索采用絮凝工艺对罗非鱼养殖尾水进行处理,选取聚合硫酸铁、三氯化铁、壳聚糖和碳酸镁等4种絮凝剂,通过比较浊度、总悬浮物和化学需氧量的去除效果,筛选出适合用于养殖尾水处理的絮凝剂。结果显示:聚合硫酸铁的效果优于其他3种絮凝剂。在投加量0.4 g/L、pH=8、絮凝时间15 min时,聚合硫酸铁对养殖尾水的浊度、总悬浮固体颗粒物(TSS)和化学需氧量(COD)的去除率分别达到了96.7%、92.6%和95.1%。对絮凝后絮体评价发现,絮体的沉降高度与浊度去除率具有一定相关性,絮体的成长受成核作用的影响。尾水处理前后的zeta电位说明聚合硫酸铁通过破坏尾水中的悬浮物的稳定结构而净化尾水。研究表明,在罗非鱼养殖尾水处理时,可选择聚合硫酸铁作为絮凝剂。

罗非鱼混养对广西三江传统稻渔综合种养系统环境及其菌群多样性的影响

研究罗非鱼混养对广西三江传统稻渔综合种养系统环境的影响,可为科学推广稻田金边鲤-罗非鱼混养模式提供基础。2021年5-10月设置3个试验组和1个对照组研究不同密度混养对种养系统环境及菌群多样性的影响,其中试验组内进行罗非鱼和金边鲤的混养,对照组为鲤单养组。结果表明,金边鲤的生长未受到罗非鱼混养的影响;高通量测序结果显示水体中的主要菌群为变形菌门(Proteobacteria)和放线菌门(Actinobacteria),其次为拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和蓝菌门(Cyanobacteria),主要功能为化能异养和有氧化能异养;底泥中的主要菌群为变形菌门和绿弯菌门(Chloroflexi),其次为酸杆菌门(Acidobacteria)、广古菌门(Euryarchaeota)和蓝菌门,主要功能类群为甲烷生成和化能异养。罗非鱼高密度混养组稻田底泥中的总氮和硝酸盐氮含量有所下降,水体中Polynucleobacter等水质良好的指示物种相对丰度增加,罗非鱼混养对稻渔综合种养系统环境及其菌群产生了有益的影响。

即食调味罗非鱼块制备工艺研究

[目的]以新鲜罗非鱼为原料制备即食罗非鱼块产品。[方法]通过单因素及正交试验以成品感官品质为评价指标对即食调味罗非鱼块的腌制去腥、调味处理、高温烘制等加工工艺进行研究。[结果]最佳工艺为:食盐4%、白酒3%、姜12%、蒜12%、葱12%腌制30分钟,食盐3%、花椒粉3%、辣椒粉6%调味30分钟,在65℃条件下烘制2h取出冷却后放冰箱冷藏24小时,65℃下复烘1h,最后刷油在160℃条件下烘烤10min。[结论]按此工艺生产的即食调味罗非鱼鱼块具有咸淡适中、肉质软硬恰当、味道鲜香麻辣的口感。

罗非鱼AT2-R抑制罗非鱼湖病毒复制的初步研究

为研究罗非鱼血管紧张素Ⅱ2型受体(AT2-R)在罗非鱼湖病毒(TiLV)感染过程中的功能,通过RT-PCR技术克隆得到体质量(20±5)g罗非鱼AT2-R(OnAT2-R)基因完整的开放阅读框序列,并运用qRT-PCR分析OnAT2-R基因的组织分布及时相表达,同时构建重组真核表达载体pEYFPOnAT2-R,在罗非鱼脑细胞中进行亚细胞定位分析,最后通过罗非鱼脑细胞过表达OnAT2-R基因后感染罗非鱼湖病毒,利用qRT-PCR方法检测TiLV-S8片段的相对表达量。试验结果显示,OnAT2-R基因开放阅读框序列全长1338bp,编码445个氨基酸,具有1个7次跨膜区域。在罗非鱼被检测的各个组织中,OnAT2-R基因均有分布,肝脏的表达量最高,其次是肌肉和皮肤,脾脏和胃的表达量最低。健康罗非鱼感染罗非鱼湖病毒后,OnAT2-R基因在肝脏中的表达量先增后减,在肌肉中的表达量先增后减再增。成功构建了pEYFP-OnAT2-R,亚细胞定位结果显示OnAT2-R主要定位于细胞膜上。在罗非鱼脑细胞中过表达OnAT2-R基因后感染罗非鱼湖病毒,发现TiLV-S8节段的mRNA水平显著降低。试验结果表明,OnAT2-R是一个潜在的定位于细胞膜且具有抑制罗非鱼湖病毒复制作用的细胞表面分子受体,这为后续深入研究罗非鱼湖病毒感染机理及罗非鱼抗病毒机制提供理论参考。

挂浆对罗非鱼片预制菜品质提升及蛋白质稳定性的影响

以罗非鱼(Oreochromis mossambicus)鱼片为原料的酸菜鱼产品是水产类预制菜的主要品种之一。为提升鱼片在复热时的质构稳定性,探讨了挂浆对鱼片品质和鱼肉蛋白稳定性的影响。通过添加淀粉、蛋清粉、花生油降低鱼片在复热过程中的蒸煮损失率,提升鱼片的质构特性,优化3类物质的添加量和工艺,以解决鱼片类预制菜在复热过程中易破碎的问题。结果表明,添加淀粉、蛋清粉和花生油均能抑制罗非鱼片的水分流失,降低蒸煮损失,提升质构特性,同时对汤汁浑浊度的影响较小。提高罗非鱼片煮制品质稳定性的最佳挂浆工艺为:淀粉添加量4.4%、蛋清粉添加量3.6%、花生油添加量1.4%、腌制时间13 min。鱼片在此条件下的综合得分(16.16±0.53)接近于预测值(16.70),说明工艺稳定可靠。与未挂浆鱼片相比,优化条件下鱼片煮制的质构硬度提高了74.2%,汤汁浑浊度降低了42.7%,蛋白质的α-螺旋和β-转角结构相对含量更多,结构更稳定,肌纤维间连接的紧密性更好,食用品质显著提升。

5种氧化型减菌剂对罗非鱼片品质及蛋白质的影响

本研究以罗非鱼片为原料,考察5种常见氧化型减菌剂(H_(2)O_(2)溶液、ClO_(2)溶液、NaClO溶液、臭氧水和微酸性电解水)的减菌效果及其对鱼片质构特性、色泽及肌原纤维蛋白(myofibrillar protein,MP)氧化情况的影响。结果表明,减菌率达80%时各氧化型减菌剂的质量浓度和作用时间分别为:1500 mg/L H_(2)O_(2)溶液浸泡8 min;200 mg/L ClO_(2)溶液浸泡10 min;200 mg/L NaClO溶液浸泡10 min;9 mg/L臭氧水浸泡10 min;30 mg/L微酸性电解水浸泡20 min。在上述5种减菌条件下对鱼肉色泽、质构特性、MP氧化情况与对照组进行比较,发现减菌处理后的罗非鱼片硬度、L*值增大,a*值减小,MP质量浓度、总巯基含量下降,羰基含量升高,表面疏水性基团数量增加,其中微酸性电解水和NaClO溶液处理组蛋白质的变性程度最明显;蛋白荧光强度降低,其中微酸性电解水处理组降低幅度最大;MP的α-螺旋和β-转角相对含量降低,β-折叠和无规卷曲相对含量增加,其中微酸性电解水处理组蛋白质二级结构变化最显著(P<0.05);MP发生不同程度的降解,微酸性电解水处理组降解程度高于其他处理组。综上所述,氧化型减菌处理后鱼肉MP发生不同程度的氧化,其中微酸性电解水处理对蛋白质的氧化作用最强。

罗非鱼皮胶原酶抑制肽的制备、体外活性及其理化特性研究

罗非鱼皮富含胶原蛋白,是一种制备生物活性肽的理想原料。以罗非鱼皮为原料,采用酶解法制备罗非鱼皮胶原酶抑制肽(Tilapia skin collagenase inhibitory peptide,TSCIP),并对其胶原酶抑制活性与金属离子结合活性的相关性进行研究,以期为罗非鱼加工副产物的高值化利用与食物蛋白源胶原酶抑制剂的研发提供参考。结果表明,罗非鱼皮经碱性蛋白酶酶解4 h后的产物具有最高的胶原酶抑制活性和锌离子(Zn^(2+))、镁离子(Mg^(2+))结合活性,并探明其主要由小分子肽(<1000 D占85.65%)组成。紫外、傅里叶变换红外光谱以及圆二色谱分析显示,TSCIP与Zn^(2+)、Mg^(2+)以及胶原酶结合后其二级结构发生了变化,β-折叠含量明显增加;Zn^(2+)、Mg^(2+)主要通过羧基氧、氨基氮原子以及羰基与TS-CIP结合,胶原酶主要通过氨基氮原子以及羰基与TSCIP结合。

植物乳植杆菌通过抑制蛋白水解改善罗非鱼发酵鱼糜凝胶强度

利用微生物发酵能有效改善淡水鱼鱼糜的凝胶强度。罗非鱼鱼糜的凝胶强度低且在加工过程中易出现凝胶劣化。为开发高品质的罗非鱼发酵鱼糜制品,以植物乳植杆菌(Lactiplantibacillus plantarum)为发酵菌株,探究了加菌发酵对罗非鱼发酵鱼糜凝胶强度的影响。结果表明,加菌发酵能够显著改善罗非鱼发酵鱼糜的凝胶强度,降低大部分游离氨基酸以及总氨基酸的含量。采用16S rRNA高通量测序技术研究加菌发酵对罗非鱼发酵鱼糜中微生物菌群的影响,结果显示,加菌发酵鱼糜中乳植杆菌属一直为优势菌属,到发酵末期增至63.71%,而其他腐败微生物菌属的丰度显著下降,同时菌落总数明显低于自然发酵鱼糜。构建了不同发酵时期的相关性网络图,结果表明在整个发酵期凝胶强度与菌落总数和总氨基酸含量呈显著负相关,仅乳植杆菌属与菌落总数和总氨基酸含量呈显著负相关,而与凝胶强度呈显著正相关。研究表明,植物乳植杆菌可以通过抑制腐败微生物的生长以及对蛋白质的水解作用来改善罗非鱼发酵鱼糜的凝胶强度。植物乳植杆菌可以作为专用发酵剂开发高品质罗非鱼鱼糜发酵制品。

ATP含量与外源酶添加对罗非鱼肌原纤维蛋白磷酸化水平的影响

蛋白质磷酸化修饰可影响肌肉品质,为探究外源酶添加对罗非鱼肌原纤维蛋白蛋白质磷酸化水平的影响,在向罗非鱼肌原纤维蛋白溶液中分别加入蛋白激酶A(protein kinase A,PKA)和碱性磷酸酶(alkaline phosphatase,AP)后进行体外孵育,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和荧光染色测定不同时间段内的磷酸化水平变化。并通过将罗非鱼肌肉浸泡在不同浓度ATP溶液中测定肌原纤维蛋白的蛋白磷酸化水平以探究ATP对其的影响。结果显示,在0~72 h,PKA组磷酸化水平均显著高于对照组和AP组,PKA组整体磷酸化水平从0 h的0.35±0.01上升至12 h的0.37±0.01,而后下降至72 h的0.29±0.01,呈先上升后下降的趋势,其中,肌球蛋白重链磷酸化水平和肌动蛋白磷酸化水平分别从0 h的0.73±0.01、0.86±0.01下降至72 h的0.58±0.02和0.68±0.01。当孵育时间为0、4、24和48 h时,3组磷酸化水平均具有显著差异。在外源添加0.3 mol/L ATP后,结果显示肌原纤维蛋白整体磷酸化水平(0.46±0.00)显著高于对照组(0.42±0.01)。PKA可促进罗非鱼肌原纤维蛋白磷酸化修饰,AP则使其去磷酸化。研究表明,宰杀后罗非鱼肌肉中的ATP含量、PKA及AP活性水平是影响蛋白质磷酸化水平的关键因素。本研究可为探明罗非鱼品质变化机制与调控策略提供理论依据。

惠州西湖外来鱼类入侵与鱼类群落状况研究

【目的】摸清惠州西湖鱼类群落组成和外来鱼类入侵现状。【方法】2023年2月,在惠州西湖主湖区开展鱼类资源调查,运用Jaccard相似性指数、相似性分析、非度量多维尺度排序法等方法分析外来鱼类物种组成、丰度差异、分布格局、群聚结构和群落相似性。【结果】共捕获鱼类21种,包括鲤、鲫和鲢等16种土著鱼类以及豹纹翼甲鲇、齐氏罗非鱼、尼罗罗非鱼、伽利略罗非鱼和露斯塔野鲮5种外来鱼类。外来鱼类质量和数量分别占总渔获物的45.36%和44.18%。豹纹翼甲鲇和齐氏罗非鱼在5种外来鱼类中质量和数量占比相对较高,表明外来种群资源量具有一定规模;相似性分析结果表明,西湖平均Jaccard相似性指数处于相似等级(数值为0.62),表明惠州西湖鱼类群落空间分布相似度高;聚类分析结果表明,在78.73%的相似性水平时,惠州西湖鱼类群落可分为4组,但它们之间的差异未达到生态学意义上的差异显著(P>0.05)。【结论】惠州西湖各站点鱼类群落较为相似,豹纹翼甲鲇和齐氏罗非鱼在各个站点均有出现,入侵较为严重,亟需采取相应的防控措施。

两个奥利亚罗非鱼品系对无乳链球菌的耐受性研究

为探讨两个奥利亚罗非鱼(Oreochromis aureus)品系对无乳链球菌(Streptococcus agalactiae)的耐受性,研究以奥利亚罗非鱼埃及品系(AE品系)和“夏奥1号”(AX品系)为研究对象,人工腹腔注射无乳链球菌,并在感染后0、7h、24h、48h、72h、120h和168h采集罗非鱼血液和脾脏,比较感染后7d存活率差异,研究血清生化指标和脾脏促炎性细胞因子表达变化。结果显示,人工感染后AE奥利亚罗非鱼存活率显著高于AX奥利亚罗非鱼。感染后两种奥利亚罗非鱼血清中的谷丙转氨酶(ALT)、谷草转氨酶(AST)、球蛋白(GLO)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)和溶菌酶(LZM)含量在感染后都显著升高;甘油三酯(TG)和白蛋白/球蛋白(A/G)显著下降,呼吸暴发受抑制;且AE品系的血清GLO和SOD水平在感染后期显著高于AX品系。定量PCR结果显示两种奥利亚罗非鱼脾脏TNF-α、IL-1β和IL-6表达在感染后7h都显著升高,但AE品系在感染后期3种促炎性细胞因子的表达都显著低于AX品系。研究表明,AE品系对无乳链球菌的抗病力强于AX品系,在感染后期的抗氧化能力更强,炎症程度更轻,从而保持对无乳链球菌更大的耐受性。研究结果为培育抗链球菌病的奥尼罗非鱼提供抗性的亲本种质资源。

RAPD-SCAR Markers for Genetically Improved NEW GIFT Nile Tilapia (Oreochromis niloticus niloticus L.) and Their Application in Strain Identification

The NEW GIFT Nile tilapia （Oreochromis niloticus niloticus L.） is a nationally certificated new strain selected over 14 years and 9 generations from the base strain of GIFT Nile tilapia, introduced in 1994. This new variety has been extended in most of areas of China. The management of genetically improved strains, including the genetic markers for identification is needed urgently. RAPD analysis was conducted and their conversion to SCAR markers was developed. From NEW GIFT Nile tilapia, two strain-specific RAPD bands, S304624 bp and S36568 bp were identified. The strain-specific RAPD bands were gel-purified, cloned, and sequenced. Locus-specific primers were then designed to amplify the strain-specific bands. PCR amplification was conducted to test the variations in allele frequencies of two converted SCAR markers among the NEW GIFT Nile tilapia and its base strains, as well as 7 additional farmed strains worldwide. The frequency of SCAR marker Ⅰ （553 bp） was 85.7% in NEW GIFT Nile tilapia, but 16.7% in the base strain. The frequency of SCAR marker Ⅱ （558 bp） was 91.4% in NEW GIFT Nile tilapia, but 0% 70% in the 7 other strains. In order to confirm the utility of these two markers, an examination was conducted for a wild population from Egypt, resulted the frequency of SCAR Ⅰ and Ⅱ was 10% and 70%, respectively, much lower than that of New GIFT strain. The increase in allele frequency of these two SCAR markers suggests that these markers might be genetically linked to the quantitative trait loci （QTL） underlining the performance traits by long term selection, and indicate the bright potential of SCAR marker technology for tracking generations during selection progress and for distinguishing among genetically improved strain and other strains.