Evaluating the use of garlic (Allium sativum) for the remedy of Cryptocaryon irritans in guppies (Poecilia reticulata)

Garlic (Allium sativum) has been shown to possess antimicrobial properties against a range of disease‐causing agents, including fish parasites. Our study aimed to investigate the potential use of garlic as a treatment against Cryptocaryon irritans infection, using guppies (Poecilia reticulata) as the fish model. Garlic was freeze‐dried, powdered, and used as it is or as an aqueous extract. The content of allicin, its main active ingredient, was 1.25 mg/g in garlic powder and 0.82 mg/ml in the aqueous extract, as determined by HPLC analysis. Aqueous garlic extract fully immobilized C. irritans theronts and protomonts within 20 and 40 min, respectively, following exposure to 5 µl/ml. Treatment trials were performed, in which infected fish were fed with garlic powder‐supplemented feeds (5%, 10%, or 20% supplementation), bathed in garlic aqueous extract (250 or 500 µl/L) and subjected to a combined treatment which included both feeding and bathing. Results revealed that the tested treatments failed to cure the infection, though reduction in infection intensity on the caudal fin, but not on the gills, was evident following dietary supplementation with 20% of powdered garlic in one of the trials.     

奶牛皱胃左方变位不同方法治疗前后瘤胃内纤毛虫数量及pH的变化

采用保守疗法、左侧腹壁切口不切开瘤胃和左侧腹壁切口切开瘤胃治疗皱胃左方变位,治疗前后不同时期对患牛进行瘤胃内纤毛虫数量及内容物pH测定.结果表明,手术治疗采用左侧腹壁切口并切开瘤胃,冲洗、取出网胃内金属异物,手术后1d瘤胃内纤毛虫数量及内容物pH较手术前1d显著下降(P＜0.05),恢复缓慢；采用左侧腹壁切口不切开瘤胃,1d后瘤胃内纤毛虫及内容物pH较手术前1d显著上升(P＜0.05),恢复较快；保守疗法瘤胃内纤毛虫数量及内容物pH变化不显著(P＞0.05)；治疗过程中纤毛虫数量受pH影响较大.     

刺激隐核虫在褐牙鲆上的传代及其生活史观察

为保存从福建省长乐市褐牙鲆上分离的刺激隐核虫、并了解其生物学特性、以褐牙鲆为宿主鱼、在水温 22~24益时、对刺激隐核虫进行传代、详细观察其生活史及各阶段的形态结构、比较不同温度处理的包囊孵化率和孵 出幼虫的感染能力、分别采集感染后2 周和4 周的褐牙鲆血清做阻动试验。目前已成功进行了40 个周期的刺激隐 核虫传代保种、每个周期8~10 d、表明褐牙鲆可作为刺激隐核虫的模式宿主鱼。观察包囊的形态结构、发现少数包囊 中存在疑似滋养体并做旋转运动的圆形原生质体。包囊5益冷藏2 个月后刺激隐核虫的孵化率下降至7.6 %、用 1 000 个5益冷藏后孵化的幼虫感染褐牙鲆、未检出滋养体。免疫血清阻动试验表明、感染后2 周和4 周的褐牙鲆血 清对幼虫的阻动效价分别为1颐320 和1颐1 280、说明体液免疫在抵抗刺激隐核虫的感染中发挥了重要作用。     

Immunization of grouper, Epinephelus coioides, confers protection against a protozoan parasite, Cryptocaryon irritans

The present study aimed to determine whether protection is conferred by immunization of grouper, Epinephelus coioides, against a protozoan parasite, Cryptocaryon irritans. The immunization of E. coioides was carried out by a low level exposure of fish to live C. irritans theronts from predetermined number of tomonts and by an intraperitoneal injection of a vaccine consisting of formalin-killed C. irritans theronts.

Mucus titers detected by ELISA were significantly higher in fingerling and adult grouper subjected to the low level of exposure to C. irritans theronts at 3-week post-exposure compared to fish that had no previous exposure. In addition, significantly smaller tomonts were produced from adult grouper after three successive exposures than the tomonts produced after a single exposure to the parasite.

In the vaccine-immunization experiment, no mortality was monitored in fish that received high dose vaccine (100 μg/fish), while 40% cumulative mortality and 100% cumulative mortality were recorded in low dose group (10 μg/fish) and control group (PBS-injected), respectively. In the succeeding replicate, the vaccine-immunized group (high dose) had 37.5% cumulative mortality and 100% cumulative mortality for the control. In addition, a total of 1830 tomonts were collected at 5-day post-challenge from the control group while none from the vaccine-immunized group. Significantly fewer trophonts and tomonts were enumerated at 5-day and 7-day post-challenge, respectively, in the vaccine-immunized group than the control.

Results suggest that a protective immunity has been conferred on the immunized grouper as indicated by high antibody titers in the mucus of C. irritans-exposed fish and higher survival and fewer parasites in vaccine-immunized fish than the control groups. The conferred immunity played a major role in preventing or limiting the adhesion, invasion, and development of C. irritans theronts on the skin of the immunized grouper.     

刺激隐核虫抑动抗原生物学信息及抗原特性分析

对刺激隐核虫抑动抗原基因进行生物学信息分析,利用抗原表位多肽制备了鼠源抗抑动抗原抗体,并研究其抗原特性.试验结果表明：利用表位串联多肽所制备的鼠源抗抑动抗原抗体包含有针对刺激隐核虫抑动抗原的特异性抗体,为刺激隐核虫单克隆抗体和疫苗的制备奠定了基础.     

人工接种反刍食团对犊牛瘤胃发育的影响

本试验采用人工方法给新生犊牛接种反刍食团,并对犊牛瘤胃内容物pH值、纤毛虫以及反刍规律等进行了研究。试验结果表明,犊牛各周龄pH值变化、接种组与未接种组之间无显著差异,但与常规组相比,在4周龄时差异则显著。接种组2周龄时发现在瘤胃中有纤毛虫,未接种组则在5周龄时发现纤毛虫,常规组在6周龄时发现纤毛虫。反刍开始时间接种组早于常规组,但各组牛的每次反刍延续时间均随着周龄的增加而延长。     

浸浴硝唑尼特对刺激隐核虫的驱虫效果

[目的]明确浸浴硝唑尼特(NTZ)对刺激隐核虫的驱虫效果,为其在水产养殖中的推广应用提供参考依据.[方法]将NTZ溶于二甲亚砜(DMSO)制成溶液,探讨NTZ对刺激隐核虫感染幼虫和滋养体的驱虫效果,并分析药物浓度、日换水量、硫酸铜(CuSO4)和渗透剂对其驱除刺激隐核虫滋养体的影响.[结果]NTZ对刺激隐核虫感染幼虫的杀灭作用随药物浓度的升高而逐渐提高,至20.0 μg/L时刺激隐核虫感染幼虫的死亡率达97.33%.经1.0 g/m3 NTZ浸浴72 h后,患病大黄鱼的刺激隐核虫滋养体完全脱落;NTZ与CuSO4配伍能有效提高药物对刺激隐核虫滋养体的驱除效果,以NTZ 1.2 g/m3+CuSO4 0.5 g/m3浸浴72 h后患病大黄鱼体表和鳃部均无刺激隐核虫滋养体寄生,病鱼存活率达100.00%;渗透剂氮酮和换水方式与NTZ驱除刺激隐核虫滋养体的效果无相关性.[结论]NTZ浸浴鱼体能有效驱杀刺激隐核虫滋养体,尤其与CuSO4配伍可有效提高药物对刺激隐核虫滋养体的驱除效果,生产中建议使用剂量为NTZ 1.2 g/m3+CuSO4 0.5 g/m3.     

鱼类特有的Ring型E3泛素连接酶MARCH5A基因在刺激隐核虫感染下的表达分析

为探究大黄鱼MARCH5A的免疫作用,实验采用反转录PCR确认了该基因的c DNA序列。该序列全长1045 bp,包含1个长度为867 bp的开放阅读框,编码288个氨基酸;序列分析显示该蛋白含1个RINGv和4个跨膜结构域。进化树分析表明大黄鱼有11个MARCH家族蛋白(与哺乳动物相同),但鱼类存在多个亚型,MARCH5A为鱼类特有蛋白,其蛋白理化性质和三维结构均与MARCH5B存在一定的差异。荧光定量PCR分析显示,MARCH5A在健康大黄鱼的多个组织中均有表达,在血液和心脏中表达量最高,其次为鳃和脑,而在肾脏和皮肤中表达量最少。刺激隐核虫感染大黄鱼后,MARCH5A在皮肤中早期表达量显著上调,第2天为对照组的9.65倍,后期下调。在鳃、脾脏和头肾中的前期表达量也有所增加,后期下降。结果表明大黄鱼MARCH5A在抗刺激隐核虫免疫应答过程中可能发挥重要作用。     

鱼类"白点病"免疫防治的研究进展

多子小瓜虫（Ichthyophthirius multifiliis）和刺激隐核虫（Cryptocaryon irritans）是两种对鱼类最具致命性的寄生性纤毛虫,分别引起淡水和海水鱼类的＂白点病＂。多年来,该病给淡水和海水养殖造成了极大的经济损失。免疫学方法是防治＂白点病＂流行极具潜力并且具有诸多优点的方法。目前尚无商业化疫苗出现。从＂白点病＂可能的疫苗开发方向出发对目前＂白点病＂的免疫防治研究现状进行综述,以期对未来的研究提供参考。