三角帆蚌瘟病组织病理学动态变化

在人工感染三角帆蚌瘟病病料组织后第3、5、7、9、11天,运用光学显微镜和电子显微镜分别观察了三角帆蚌(Hyriopsis cumingii)外套膜、鳃和斧足的病理变化特征.结果表明,三角帆蚌瘟病病毒对3种器官组织有不同程度的破坏作用,外套膜和鳃的病变比较严重,斧足稍轻.光镜下,前5 d内各组织结构变化不大;从第7天开始,出现上皮组织细胞肿胀、排列紊乱、部分脱落甚至完全坏死等显著的病理变化.电镜下,前5 d细胞结构无明显病理变化,只出现了少量的病毒粒子;从第7天开始,病毒粒子数量增加,线粒体、内质网、核糖体等细胞器解体,细胞空泡化,但斧足的超微结构变化不明显.     

大口黑鲈致死性结节病病原的分离、鉴定及组织病理学观察

2018年4月四川邛崃地区某养殖场大口黑鲈感染致死性结节病,死亡率高达80%。为明确病因,通过常规微生物分离、鉴定,从患病鱼皮肤溃疡灶、鱼鳔腔积液和肝脏组织中分离到同一株优势菌,命名为HSY-NS02。经菌体形态观察、革兰氏染色和抗酸染色镜检、生理生化实验、16S rDNA序列扩增及系统发育分析和特异性PCR扩增,确定该菌为诺卡氏菌。进一步对健康大口黑鲈进行分离菌的人工感染实验以确定其致病性,结果显示,人工感染鱼出现与自然发病相似症状,且从人工感染鱼体中再次分离到相同菌。组织病理学观察显示,皮肤溃疡灶、心脏、肝脏、脾脏、肾脏和鳃发生不同程度的慢性炎性肉芽肿病变,其中脾脏病变最为严重。对菌株HSY-NS02进行药物敏感性实验,结果显示,该菌对庆大霉素、新霉素和制霉菌素3种药物敏感,对其他18种药物耐受,呈多重耐药性。     

Effect of using Spirulina and Chlorella as feed additives for elevating immunity status of Nile tilapia experimentally infected with Aeromonas hydrophila

The present trial was performed to evaluate the effect of dietary incorporation of dried Spirulina, Chlorella and their mixture on the immune status of Nile tilapia (Oreochromis niloticus) before and after artificial infection with pathogenic bacterium Aeromonas hydrophila (A. hydrophila). Two hundred and forty fish were divided into four groups: (a) a control group fed on a basal diet only, (b) a second group fed on a diet containing Spirulina 15%, (c) a third group fed on a diet containing Chlorella 15%, and (d) a fourth group fed on a diet containing a mixture of both Spirulina 15% and Chlorella 15%. At the end of the experiment (9 weeks), the four groups were experimentally infected with A. hydrophila for 7 days. Antioxidant enzymes, lysozyme and bactericidal activities and histopathological changes were determined just before the challenge test and 7 days post‐challenge. Significant (p ≤ 0.05) increases in fish body protein% before the challenge test and increases in serum antioxidant enzymes, lysozyme and bactericidal activity in the Chlorella and algal mixture groups before and after the challenge test were observed. Spirulina, Chlorella and their mixture groups significantly decreased serum malondialdehyde compared to the control group before and after the challenge test. Using Spirulina, Chlorella and their mixture mitigated the necrotic and degenerative changes induced by A. hydrophila and revealed well‐developed and multiple melanomacrophage centres. Thus, dietary Spirulina, Chlorella and their mixture inclusion in Nile tilapia fish proved to have a protective effect against A. hydrophila infection.     

Pancreas disease (PD) in sea‐reared Atlantic salmon,Salmo salar L., in Norway; a prospective,longitudinal study of disease development and agreement between diagnostic test results

A prospective longitudinal study was performed on three cages at each of three Norwegian Atlantic salmon seawater sites that experienced outbreaks of pancreas disease (PD). Once salmonid alphavirus (SAV) ribonucleic acid (RNA) was detected by real‐time RT‐PCR (Rt RT‐PCR) at a site, it became detected in all studied cages and was persistently found until the end of the study period up to 19 months after first detection. SAV‐specific antibodies were detected at all sites until the end of the study period and were also found at a high prevalence in broodfish at the time of stripping. No evidence of increased viral activity was detected in these broodfish. One site tested negative over several months prior to the first detection of SAV by Rt RT‐PCR and SAV‐specific antibody, which occurred 1 month prior to clinical manifestations of PD. Moribund fish or thin fish/runts that were sampled after the first PD diagnosis had almost twice the risk of testing positive by one or more diagnostic tests compared to that of randomly selected apparently healthy individuals. This paper describes the first detailed investigation of the disease development of PD at site and cage level in Norway, as well as an assessment of the performance and agreement of the commonly used diagnostic tests.     

大黄鱼感染哈维氏弧菌后血液生化指标的变化及组织病理学观察

从患病大黄鱼病灶中分离出病原哈维氏弧菌,并人工回接感染大黄鱼,病鱼出现典型的皮肤溃疡症。测定大黄鱼血液各项指标发现,病鱼单核细胞、嗜中性粒细胞、血栓细胞百分率明显下降,而淋巴细胞百分率增加,均呈极显著差异(P<0.01)。患病大黄鱼总蛋白为(12.8±3.95)g/L,对照组鱼为(19.4±4.33)g/L,病鱼球蛋白、甘油三脂、血糖含量也明显低于对照组(P<0.01)。然而病鱼谷丙转氨酶为(37±5.11)IU/L,对照组为(28±5.54)IU/L;病鱼谷草转氨酶为(164±47.19)IU/L,对照组为(117±21.84)IU/L,均呈极显著差异(P<0.01)。研究结果说明,病鱼肝脏和肾脏组织受损。对患病大黄鱼进行病理组织学观察,结果发现病鱼病变组织头肾、肝出现不同程度的变性以及坏死。细胞超微病变观察发现,头肾肾小管上皮细胞变性、坏死。肝细胞胞浆内脂滴积累多。血清指标的测定可以作为大黄鱼哈维氏弧菌病细菌感染的指标。     

胆碱对高脂胁迫的团头鲂肝脏抗氧化、组织结构和免疫力的影响

以初始体质量(42.20±0.34)g团头鲂为研究对象,研究饲料中添加胆碱对高脂胁迫团头鲂肝脏抗氧化、组织结构和免疫力的影响。实验采用2×2因子设计方法,共配置2个脂肪水平(5%和15%)和2个胆碱水平(1200和2200 mg/kg)的等氮饲料4组。养殖实验在室内循环水系统中进行,养殖周期为8周。养殖实验结束后进行血液和肝脏生化指标、肝脏抗氧化指标和非特异性免疫指标等的检测分析,以及观察肝细胞的超微结构变化。结果显示,当饲料中添加1200 mg/kg胆碱时,随着脂肪水平的升高,团头鲂肝脏脂肪含量和甘油三酯浓度显著增加,而血浆中谷草转氨酶(AST)和谷丙转氨酶(ALT)活性则显著上升;肝脏中丙二醛(MDA)含量、总超氧化物歧化酶(t-SOD)活性、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX)活性等显著升高;血液中白细胞数、溶菌酶活性、总蛋白和球蛋白水平以及还原型谷胱甘肽(GSH)活性显著降低;超微结构观察到肝细胞中存在大量脂滴,伴随着细胞核变性、核膜残缺、线粒体水肿等异常现象。这些结果表明高脂日粮不仅导致肝细胞的结构和功能的损伤,而且造成肝脏氧化应激和机体免疫力的下降。在高脂饲料(15%脂肪)中添加2200 mg/kg胆碱,可显著降低肝脏脂肪含量、血浆谷草转氨酶(AST)和谷丙转氨酶(ALT)活性,显著降低肝脏丙二醛(MDA)含量、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,并显著提高白细胞数和球蛋白水平以及还原型谷胱甘肽(GSH)活性,同时肝细胞形态及细胞器结构也趋于正常。研究表明,添加适量胆碱能够减少肝脏脂肪沉积,维持肝脏结构和功能的正常,并增强团头鲂抗氧化能力和机体免疫力,继而保持鱼体的健康。     

蟾蜍似矛球吻棘头虫的临床病例研究

【目的】 对1例中华蟾蜍多脏器寄生虫感染临床案例进行病理诊断并通过分子生物学鉴定其物种分类。【方法】 对实验室饲养的1只消瘦、拒食后发生死亡的雌性中华蟾蜍进行解剖观察，病理组织学研究，采用蠕虫剖检法、结节压片法对分离获得的虫体进行形态学观察研究，并进行分子生物学鉴定。【结果】 剖检发现蟾蜍心脏、肝脏等脏器及多系膜寄生棘头虫，未在蟾蜍体内发现线虫、吸虫等其他蠕虫寄生。HE染色结果显示，棘头虫的感染寄生造成了蟾蜍肺脏、肝脏等多脏器的机械性损伤、炎性反应，其中肠壁黏膜发生严重损坏脱落。通过PCR扩增其细胞色素c氧化酶亚基Ⅰ（cytochrome c oxidase subunit Ⅰ，COⅠ）基因片段，获得5株不同部位的目的虫株COⅠ基因目的片段，经对比发现，鉴定的棘头虫均为同一种棘头虫，且其GC含量在40.14%~40.83%之间。选取JS-01株进行NCBI-BLAST比对，与似矛球吻棘头虫（Sphaerirostris lanceoide，登录号：MG931943.1）相似性最高，为99.5%。借助分子生物学分析并结合形态学观察结果确定其为似矛球吻棘头虫。【结论】 似矛球吻棘头虫的大量感染寄生造成了蟾蜍的死亡，本试验结果可为药用蟾蜍棘头虫病的防控提供临床症状、病理、分子生物学等方面的参考，对两栖动物似矛球吻棘头虫的诊断与研究具有参考价值。     

脊尾白虾“僵尸病”的初探

为确定2018年冬季以来江苏省沿海地区养殖脊尾白虾患“僵尸病”的病原及流行病学特点，实验采用LB培养基和PDA培养基从病虾血淋巴中分离得到直径为1~3 mm、边缘整齐、米黄色隆起菌落；人工回感实验结果显示，回感后的脊尾白虾表现出与自然患病脊尾白虾相同的症状，并在回感脊尾白虾体内也分离出了相同的菌株，符合科赫氏法则。对该菌株进行形态观察结合18S rRNA序列对比及系统发育分析，发现分离菌株MQ2101具有酵母的典型形态，且与二尖梅奇酵母相似度达99.82%，结果表明菌株MQ2101为二尖梅奇酵母。致病性结果初步分析显示，MQ2101对脊尾白虾的半致死浓度 (LD₅₀)为1.39×10⁷ CFU/尾。病理学观察发现，患病脊尾白虾鳃、肌肉和肝胰腺均发生不同程度的病变，其中肝胰腺病变最为严重，肝小管呈现空泡化，管腔体积变大；在鳃和肝胰腺组织中均存在大量定殖的菌体。流行病学调查结果显示，每年2—5月发病迅速，发病率为5%~30%，死亡率为3%~10%。本研究确定了二尖梅奇酵母为江苏沿海地区脊尾白虾“僵尸病”的病原，其对脊尾白虾具有较强致病性，主要侵染组织为肝胰腺和鳃。以上研究结果为脊尾白虾“僵尸病”的防控提供了相关科学依据。