3种海参的主要活性成分和免疫调节作用的比较研究

采用化学分析和免疫学评价方法，分析比较了美国肉参（Isostichopus badionotus）、墨西哥刺参（Isostichopus fuscus）和冰岛刺参（Cucumaria frondosa）的主要活性成分及其对小鼠免疫功能的调节作用。结果表明，3种海参体壁中以冰岛刺参的皂苷含量最高，墨西哥刺参的黏多糖含量最高。冰岛刺参体壁总氨基酸、药效氨基酸和鲜味氨基酸总量均为最高。对于正常小鼠，3种海参均能提高小鼠胸腺／体质量比值（美国肉参，P〈0．05）、增加足跖厚度（P〈0．05或P〈0．01）和促进巨噬细胞对血液中碳粒的吞噬指数（P〈0．05或P〈0．01）。对于免疫机能低下模型小鼠，3种海参均能显著提高小鼠胸腺／体质量比值（P〈0．05或P〈0．01）、促进小鼠脾淋巴细胞的增殖和转化（P〈0．05或P〈0．01）、提高小鼠血清溶血素含量（P〈0．05或P〈0．01）和巨噬细胞对血液中碳粒的吞噬指数（P〈0．05或P〈0．01）。提示3种海参均具有显著的免疫调节作用。对于正常小鼠，主要以促进细胞免疫和非特异性免疫途径为主。对于免疫机能低下小鼠，3种海参均能提高机体的体液免疫、细胞免疫和非特异性免疫功能。其中，冰岛刺参在促进体液免疫和非特异性免疫方面的作用更突出，可能与其含有较高含量的皂苷和免疫活性氨基酸有关。     

鲟鱼硫酸软骨素对小鼠免疫器官肥大细胞的影响

为研究鲟鱼硫酸软骨素对小鼠免疫器官的影响,连续对小鼠灌胃不同剂量鲟鱼硫酸软骨素2周后,外科手术取出胸腺和脾脏,并依次进行称重、卡洛氏固定、石蜡包埋、组织切片、甲苯胺蓝染色,40倍物镜下观察鲟鱼硫酸软骨素对小鼠免疫器官肥大细胞数量的影响。结果表明：鲟鱼硫酸软骨素能促进小鼠免疫器官重量指数增大,并能显著促进免疫器官中肥大细胞数量的增长。其中,600 mg/kg剂量下小鼠胸腺指数显著( P<0．05)增大,胸腺中肥大细胞数量极显著( P<0．01)增加,60 mg/kg剂量下小鼠脾脏重量的增加最为明显( P<0．01),6 mg/kg剂量下小鼠脾脏中肥大细胞数量的增加最为明显( P<0．01)。     

维生素A缺乏对建鲤生长性能及免疫功能的影响

幼建鲤体重(11 37±0 55)g,投喂不添加或添加维生素A的3组试验饲料,进行饲养实验70d,攻毒实验15d和免疫接种实验24d,研究维生素A缺乏对幼建鲤生长性能和免疫功能的影响,结果表明,维生素A缺乏组(0IU/kg)建鲤的成活率、摄食量、增重、净蛋白质沉积率、肥满度、肝胰脏指数、肝胰脏维生素A含量、肠道皱襞高度,头肾重量、脾脏重量和指数、后肾重量和指数、血液红细胞、白细胞数量和血清溶菌酶活力显著降低(P<0 05),且饲料系数显著增加(P<0 05),而对头肾体指数、用灭活嗜水气单胞菌免疫后第10和17天血清抗体水平影响不显著(P>0 05)。基于本实验研究结果,可知维生素A是维持幼建鲤正常生长和免疫机能必不可少的营养成分。当饲料中维生素A含量达到3969IU/kg时,一定程度上可满足幼建鲤正常生长和免疫机能的需要,进一步提高饲料维生素A含量(达23816IU/kg),对幼建鲤生长和饲料利用没有显著影响,但在一定程度可提高其免疫机能。     

脂肪酶对南方鲇生长性能、消化酶活性及血液生化指标的影响

在基础饲料中分别添加0、0.1、0.3 g/kg的脂肪酶,饲喂平均体重(57.6±0.02)g的南方鲇(Silurus meridionalis Chen)60 d,测定南方鲇的生长性能、饲料表观消化率、消化酶活性及血液生化指标。结果显示:添加0.1 g/kg和0.3 g/kg的脂肪酶,增重率比对照组分别提高了4.0%和5.8%,但差异不显著(P>0.05),饲料系数分别下降了5.0%和6.6%,差异显著(P<0.05),肥满度和粗脂肪消化率显著提高(P<0.05),对内脏指数、肝脏指数、肠脂指数、胰蛋白酶活力、肠蛋白酶活力、肠脂肪酶活力及血液生化指标均没有显著影响(P>0.05)。添加0.3 g/kg的脂肪酶显著提高胰脂肪酶活力(P<0.05),添加0.1 g/kg的脂肪酶对胰脂肪酶活力没有显著影响(P>0.05)。结果表明,在脂肪水平为10%的饲料中添加0.3 g/kg的脂肪酶可以提高胰脂肪酶活力,提高脂肪消化率,改善饲料利用率。     

pH对栉孔扇贝体内几种免疫因子的影响

测定不同pH（7．0，7．3，7．6，8．5，9．0）环境胁迫下栉孔扇贝（Chlamys farreri）体内酸性磷酸酶（ACP）、碱性磷酸酶（ALP）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）及胞内活性氧含量（ROIs）等5种免疫指标的变化。结果表明，在pH高于正常组（pH8．0）时，上述5种免疫指标均随pH的增加而明显升高；在pH低于正常组时情况稍复杂，4种酶活性均呈先上升后下降的趋势。ACP在pH7．6时酶活性上升至峰值，随后酶活性降低；ALP、SOD和CAT活性均在pH7．3时达峰值，随后酶活性降低；而胞内活性氧含量则随pH降低持续增加。实验结果表明，pH对栉孔扇贝免疫活性有明显的刺激作用，且在一定范围内随刺激强度的增加呈现免疫活性的正调节；但在高强度pH刺激下，扇贝免疫活性呈现出负调节，这种负调节可能与高强度刺激引起的免疫机能疲劳或损伤有关。[中国水产科学，2006，13（4）：650—654]     

Cu↑（2+）对栉孔扇贝体内几种免疫因子的影响

测定了栉孔扇贝(Chlamys atreri)在不同浓度Cu^2 胁迫下酸性磷酸酶(ACP)、碱性磷酸酶(AKP)及过氧化氢酶(CAT)等3种酶活性的变化,并同时用组织化学的方法检测了其对栉孔扇贝活性氧(R0S)含量的影响。结果表明,当Cu^2 在0．02mg／L、0．05mg／L、0．1mg／L3种质量浓度条件下,栉孔扇贝体内ACP和AKP活性均高于对照组,但随着Cu^2 浓度增加,酶活性有降低的趋势,当Cu^2 质量浓度达到0．2mg／L时这2种酶活性均低于对照组。而CAT活性均高于对照组,且随着Cu^2 质量浓度增加而升高;活性氧含量的变化趋势与CAT活性变化趋势相似。这些数据说明Cu^2 对栉孔扇贝的免疫活性有明显影响。     

裙带菜褐藻酸钠的分离纯化及降血脂作用的研究

为探讨裙带菜褐藻酸钠对饮食性高血脂的影响,从裙带菜中提取褐藻酸钠,经DEAE-Sepharose FF、Sepharose 4B纯化；以高脂乳剂对小鼠灌胃,建立高脂血症模型,再给小鼠灌胃褐藻酸钠溶液,研究裙带菜褐藻酸钠对小鼠血脂水平的影响。结果表明,与正常对照组相比,模型对照组小鼠血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和动脉硬化指数(LDL-C/HDL-C)水平显著升高,高密度脂蛋白胆固醇(HDL-C)和抗动脉粥样硬化因子(HDL-C/TC)水平显著降低,成功建立起高血脂模型；与模型对照组相比,试验组小鼠血清的TC、TG、LDL-C和LDL-C/HDL-C水平显著降低,HDL-C和HDL-C/TC水平显著升高。结果表明,裙带菜褐藻酸钠具有较强的降血脂作用。     

大菱鲆消化酶的活力

以1龄大菱鲆(Scophalmus maximus)为研究材料，研究养殖水温下大菱鲆消化酶的分布及不同温度和不同pH条件下胃、幽门盲囊、前肠、中肠、后肠中蛋白酶、淀粉酶、脂肪酶的活性。确定饵料中主要营养成分的消化部位和主要消化酶的性质。结果表明，各部位蛋白酶活性的最适温度和pH依次为40℃，2．0；40℃，8．0；60℃，8.0；40℃，8.5；60℃，8．0。18℃养殖水温下，在适宜的pH范围内，各部位蛋白酶活力从高到低依次为幽门盲囊、胃、前肠、中肠、后肠；淀粉酶活性的最适温度和pH依次为40℃，6．5；40℃，8．0；40℃，7．5；50℃，7．0；40℃，8．0；淀粉酶活力从高到低依次为幽门盲囊、前肠、中肠、后肠、胃；脂肪酶活性的最适温度和pH依次为40℃，7．5；40℃，6．5；40℃，7.0；40℃，7.5；40℃，7.5；脂肪酶活力从高到低依次为前肠、中肠、幽门盲囊、后肠、胃。     

"翻背症"对方斑东风螺主要消化酶及免疫相关酶的影响

为探索方斑东风螺消化及免疫系统对"翻背症"的应答机制,采集了健康(对照组)和"翻背症"早期、中期和晚期的方斑东风螺,采用TCBS平板计数法检测螺体内弧菌含量,试剂盒法检测螺体内组织超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、溶菌酶(LZM)等免疫相关酶和胃蛋白酶、脂肪酶、淀粉酶等消化酶活性。结果显示,方斑东风螺体内弧菌含量随病情的发展而显著增加,晚期达到最大值。发病早期,CAT、SOD、POD、ACP和胃蛋白酶等酶活性均显著上升。发病中期,POD、AKP、LZM活性均上升,且显著高于对照组和发病早期;与发病早期相比,CAT、SOD、ACP、胃蛋白酶、脂肪酶活性有所下降;与对照组相比,CAT、ACP活性仍显著升高,胃蛋白酶、脂肪酶活性显著降低,而SOD、淀粉酶活性无显著变化。发病晚期,各消化酶与免疫相关酶活性均较发病中期有所下降,但POD、ACP活性仍显著高于对照组,CAT、SOD、AKP、LZM活性与对照组无显著差异,而胃蛋白酶、脂肪酶、淀粉酶活性显著低于对照组。研究表明,"翻背症"方斑东风螺体内弧菌的暴发过程中,机体的免疫系统与消化系统均参与了机体病害免疫应答反应,且可以采用CAT、POD和ACP活性作为方斑东风螺"翻背症"的检测评价指标。     

复合益生菌对中华鳖生长及血液生化指标的影响

为了探究饲料中添加复合益生菌对中华鳖（Trionys sinensis）生长及血液生化指标的影响,在体重为（227．47±14．42）g 的中华鳖基础饲料中分别添加复合益生菌0 mg／kg（对照组Ⅰ）、500 mg／kg（试验组Ⅱ）、1000 mg／kg （试验组Ⅲ）和2000 mg／kg（试验组Ⅳ）;试验共分4组,每组3个重复,在（30±2）℃温室中进行,投喂140 d。结果表明,各试验组均能显著提高中华鳖增重率、特定生长率和存活率（P ＜0．05）,降低饵料系数（P ＜0．05）,且Ⅲ组增重率、特定生长率和存活率最高,饵料系数最低,增重率比Ⅰ组提高了29．25％;各试验组均能显著提高中华鳖脂肪酶、淀粉酶和胰蛋白酶活力（P ＜0．05）,当添加量为2000 mg／kg 时,淀粉酶活力有所下降（P ＜0．05）;与对照组相比,各试验组均能显著降低血清中总胆固醇、尿素氮水平和谷丙转氨酶活力（P ＜0．05）,提高甘油三酯含量（P ＜0．05）,而血清中总蛋白、球蛋白、白蛋白、血糖、丙二醛含量和谷草转氨酶活力差异不显著（P ＞0．05）;各试验组均能显著提高血清中溶菌酶、谷胱甘肽过氧化物酶和超氧化物歧化酶活力（P ＜0．05）,但过量添加（2000 mg／kg）会降低其活力（P ＞0．05）。在基础饲料中适量添加复合益生菌能促进中华鳖的生长、降低饵料系数、提高肠道消化酶活性,同时还具有提高非特异性免疫和抗氧化功能,最适添加量为 1000～2000 mg／kg。     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Protein and amino acid composition from the mantle of juvenile O ctopus vulgaris exposed to prolonged starvation

Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg^?1 DW, and it decreased with increasing fasting days. The non‐essential amino acids content was higher (486.18 ± 11.08 g kg^?1 protein) than essential amino acids (425.82 ± 9.15 g kg^?1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.