几株益生菌对鲤前肠黏液的体外黏附特性

采用体外固定鲤前肠黏液,结合细菌同位素示踪的方法,对来源于鲤肠道的2株肠球菌和1株柠檬酸杆菌以及养殖水体的2株芽孢杆菌的细菌表面凝集素和黏液蛋白受体的化学组成、对病原菌附着鲤前肠黏液的影响等进行了研究。结果表明,5株菌经高碘酸钠和蛋白水解酶修饰后,高碘酸钠能极显著降低芽孢杆菌、肠球菌和柠檬酸杆菌的黏附率(P<0.01),而蛋白水解酶对多数菌株的影响不显著(P>0.05),推测细菌表面的凝集素主要为具糖蛋白性质的物质。黏液蛋白经蛋白水解酶处理后,部分菌株的附着数量显著降低(P<0.05),但经高碘酸钠处理后,5株菌的黏附率均显著上升(P<0.05),提示黏液蛋白上存在的5株细菌的特异性受体物质可能为蛋白质。益生菌通过排斥、竞争和取代作用能够显著降低部分病原菌的黏附率。取代作用对病原菌黏附的抑制效果最好,但具有菌株特异性。     

鲤益生茵筛选及部分菌株对鲤前肠黏液的体外黏附作用

从养殖池、水族箱和健康鲤肠道等分离20株细菌,通过耐热和体外拮抗试验筛选出5株细菌,经生化试验和细菌:16SrDNA测序鉴定为2株芽孢杆菌(Bacillus)、2株肠球菌(Enterococci)和1株柠檬酸杆菌(Citrobacter).进一步采用体外固定鲤前肠黏液蛋白,结合同位素32P标记细菌并示踪的方法,研究来源于鲤肠道的肠球菌和柠檬酸杆菌以及鲤养殖水体中的芽孢杆菌对鲤前肠黏液的体外黏附活性,建立筛选鲤(Cyprinus carpio)益生菌的方法.结果表明,5株细菌均能黏附到黏液体外模型,肠球菌的相对黏附率极显著高于芽孢杆菌与柠檬酸杆菌(P<0.01),柠檬酸杆菌与芽孢杆菌的相对黏附率差异不显著(P>0.05),而肠球菌L2的相对黏附率极显著高于肠球菌E2(P<0.01).研究证明,鱼源的肠球菌对鲤前肠黏液的黏附率高于异源的芽孢杆菌,而且不同种属的肠球菌在黏附能力上也存在差异.     

鲤益生菌筛选及部分菌株对鲤前肠黏液的体外黏附作用

从养殖池、水族箱和健康鲤肠道等分离20株细菌,通过耐热和体外拮抗试验筛选出5株细菌,经生化试验和细菌16S rDNA测序鉴定为2株芽孢杆菌（Bacillus）、2株肠球菌（Enterococci）和1株柠檬酸杆菌（Citrobacter）。进一步采用体外固定鲤前肠黏液蛋白,结合同位素^32P标记细菌并示踪的方法,研究来源于鲤肠道的肠球菌和柠檬酸杆菌以及鲤养殖水体中的芽孢杆菌对鲤前肠黏液的体外黏附活性,建立筛选鲤（Cyprinus carpio）益生菌的方法。结果表明,5株细菌均能黏附到黏液体外模型,肠球菌的相对黏附率极显著高于芽孢杆菌与柠檬酸杆菌（P〈0.01）,柠檬酸杆菌与芽孢杆菌的相对黏附率差异不显著（P〉0.05）,而肠球菌L2的相对黏附率极显著高于肠球菌E2（P〈0.01）。研究证明,鱼源的肠球菌对鲤前肠黏液的黏附率高于异源的芽孢杆菌,而且不同种属的肠球菌在黏附能力上也存在差异。     

鳜皮肤黏液IgM样蛋白的纯化

对经嗜水气单胞菌全菌疫苗浸泡免疫后的鳜皮肤黏液中的免疫球蛋白采用盐析法、重组蛋白A(HiTrap rProteinA Sepharose)亲和层析法及鼠抗鳜血清IgM单克隆抗体偶联的Sepharose 4B亲和层析法分离纯化,并通过SDS-PAGE及Western-blot技术对纯化蛋白的部分特性进行分析比较。结果表明:50%硫酸铵溶液可以沉淀黏液中大部分蛋白,条带仍较多,约十几条,其中含有72 ku和29 ku的条带,因此仅可作为免疫球蛋白粗提的方法;Sepharose 4B亲和层析法所提取鳜黏液Ig经SDS-PAGE检测,只含有72 ku和29 ku 2个条带(初步认为鳜黏液Ig的重链和轻链),与鳜血清IgM的重、轻链分子量相同;rProteinA亲和层析法所提蛋白除具有上述重链(72 ku)和轻链(29 ku)外,还含有43 ku的蛋白带(可能为鳜黏液Ig另外一种形式的重链)。Western-blot显示,兔抗鳜Ig多克隆抗体可与72 ku及43 ku条带发生发应。两种亲和法所提蛋白纯度较高,但含量较低,条带较淡,仅可作为实验室小量提纯鳜黏液Ig的有效方法。     

一株广温性大菱鲆肠道益生菌的筛选与鉴定

从健康的大菱鲆肠道内分离细菌,以大菱鲆致病菌鲨鱼弧菌Vibrio archariae和大菱鲆弧菌Vibrio scophthalmi为指示菌,根据拮抗作用原则,分离得到一株菌TYTG-1.根据菌株的形态、生理生化特征和16S rDNA序列分析,菌株被鉴定为枯草芽孢杆菌Bacillus subtilis.从养殖大菱鲆肠道中分离获得枯草芽孢杆茵在国内属首次报道.该株茵在4～42℃均能生长,属广温性.经过28d的投喂添加浓度为107和109CFU/g枯草芽孢杆菌的饲料,大菱鲆生长正常,未产生不良反应.该株枯草芽孢杆茵对引起大菱鲆腹水病和肠炎病的两株致病弧茵具有良好的拮抗作用,其具有较大潜力作为肠道益生菌应用于大菱鲆的养殖.     

红鱼粉饲料中添加海洋红酵母对大菱鲆生长性能的影响

为研究用红鱼粉替代白鱼粉,并在替代饲料中添加两个不同梯度海洋红酵母对大菱鲆生长性能、体成分以及形体指标的影响。试验用白鱼粉、红鱼粉、海洋红酵母1 g／kg＋红鱼粉、海洋红酵母2 g／kg＋红鱼粉4种等氮等能的试验饲料分别饲喂初始体质量（213．48±0．23） g的大菱鲆,480尾试验鱼随机分为4组,每组设3个重复,每个重复40尾,试验期为6周。试验结果表明,与红鱼粉组相比,海洋红酵母1 g／kg＋红鱼粉组可显著提高大菱鲆的摄食率、相对质量增加率、特定生长率及蛋白质效率（P＜0．05）,效果与白鱼粉组相当,高于海洋红酵母2 g／kg＋红鱼粉组,但差异不显著（P＞0．05）。各试验组大菱鲆的体成分和形体指标差异不显著（ P＞0．05）。据此,添加海洋红酵母1 g／kg即能显著提高大菱鲆对红鱼粉替代白鱼粉饲料的摄食率、蛋白效率和生长性能,使得红鱼粉替代组的饲料性能达到了白鱼粉组的生长水平。     

大菱鲆致病性溶藻弧菌SR1的外膜蛋白及其抗原性分析

用十二烷基肌氨酸钠(Sarkosyl)抽提结合超速离心的方法提取了一株大菱鲆致病性溶藻弧菌(Vibrio alginolyticus)SR1和其他7株弧菌的外膜蛋白。通过SDS-PAGE图谱分析比较了这8株弧菌外膜蛋白的组成,结果表明,8株弧菌的外膜蛋白电泳一般可得到6-12条条带,其分子量多集中在65-106 kD和28-48 kD,其中36 kD的蛋白带为8株弧菌所共有。用兔抗SR1全菌血清进行Western-blot印迹显示,菌株SR1的外膜蛋白条带中有6条发生了阳性反应,其分子量分别为73 kD、48 kD4、5 kD3、9 kD、36 kD和32 kD。而其他7株弧菌的外膜蛋白与兔抗SR1血清也发生程度不等的阳性反应,这些阳性反应条带的分子量集中在65-73 kD、45-48 kD和36-41 kD之间,其中36 kD的外膜蛋白在8株弧菌中均出现明显的阳性反应,说明36 kD的外膜蛋白是这8株弧菌共有的特异性抗原。     

益生菌在水产动物肠道内黏附机制的研究进展

双歧杆菌、乳酸杆菌、芽孢杆菌、酵母是益生菌制剂中最常用、最具代表性的菌种。大多数商品化的益生菌制剂是将这些菌以单一的或复合的形式,混合在粉料载体中或添加些蛋白性原料,然后将益生菌制剂直接混合在饲料中添加或制成混合液直接加入水体。各种菌通过肠道并在肠道壁黏附发挥益生作用,能否黏附定植于肠道壁是益生菌作用的前提,也是菌种筛选的重要指标。已有的研究表明,各类微生物的黏附是非特异性的,即与菌体的结构有关,菌体能接近肠道上皮细胞,菌体的特殊配体进一步与宿主细胞相应的受体之间特异性结合,既黏附素一受体学说。     

鲢鱼骨骼肌过敏原小清蛋白的分离纯化及多克隆抗体制备与应用

小清蛋白是鱼类的主要过敏原,对该蛋白的研究不仅有利于过敏原检测方法的建立也可为低致敏性水产品的开发提供理论依据。通过组织捣碎、冷冻离心、热处理、Superdex75凝胶过滤等方法从鲢白色肉中纯化得到过敏原小清蛋白。Tricine-SDS-PAGE显示,在非还原条件下,该蛋白呈分子量分别为12ku、14ku、24ku的3个条带。而在还原条件下,仅有分子量为12ku的条带。Western-blotting分析表明,分子量为12ku、14ku和24ku的这3个条带都与小鼠抗蛙小清蛋白单克隆抗体(PARV-19)发生特异性反应,提示它们均为小清蛋白的不同形态。用纯化的小清蛋白制备多克隆抗体,经Protein A Sepharose亲和层析纯化得到高纯度的免疫球蛋白G(IgG)。Dot-blot检测发现,抗体稀释至1/51200时仍能与纯化的小清蛋白有显色反应。用制备的多克隆抗体进行Western-blotting分析,能特异地检测4种鱼(鲤、鲢、鲫、黄鳍鲷)中的小清蛋白。     

鳗鲡肠道菌株筛选及菌株特性研究

从鳗鲡肠道中分离获得4株优势菌株,通过16S r RNA测序鉴定,结果分别为植物乳杆菌L2、鼠李糖乳杆菌L4、地衣芽孢杆菌B1与枯草芽孢杆菌B2。对这4株菌株进行特性研究,结果表明,这4株益生菌对副溶血弧菌、铜绿假单胞菌、鳗弧菌等具备较好的抑菌能力,且能高效降解氨氮、亚硝酸盐;对青霉素、氯霉素、庆大霉素等8种药物表现敏感,在5%兔血琼脂平板培养无溶血环形成,对鳗鲡无致病性。指出,筛选出来的这4株鳗鲡肠道益生菌,可为后续益生菌制剂的研发提供理论依据,为鳗鲡安全、环保养殖新增一条可持续发展道路。     

Interactions of microorganisms isolated from gilthead sea bream, Sparus aurata L., on Vibrio harveyi, a pathogen of farmed Senegalese sole, Solea senegalensis (Kaup)

Four bacterial isolates from farmed gilthead sea bream, Sparus aurata, included in a previous study as members of the Vibrionaceae and Pseudomonodaceae and the genus Micrococcus, have been evaluated for their adhesive ability to skin and intestinal mucus of farmed Senegalese sole, Solea senegalensis, and their antagonistic effect on Vibrio harveyi, a pathogen of sole. These isolates showed higher adhesion to sole mucus than the pathogenic strains of V. harveyi assayed. Only two of the isolates showed antagonistic activity to V. harveyi. Interactions of the four isolates with V. harveyi in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions were studied. Three isolates were able to reduce the attachment to skin and intestinal sole mucus of a pathogenic strain of V. harveyi under displacement and exclusion conditions, but not under competition conditions. The in vivo probiotic potential of isolate Pdp11 was assessed by oral administration followed by challenge with the pathogenic V. harveyi strain Lg14/00. A group of 50 Senegalese sole received a commercial diet supplemented with 10(8) cfu g(-1) of lyophilized Lg14/00 for 15 days. A second group of fish received a non-supplemented commercial diet. After challenge the mortality of the fish receiving the diet supplemented with the potential probiotic isolate was significantly lower than that in the fish receiving the non-supplemented commercial diet. This study has shown that the ability to interfere with attachment of pathogens, as well as the adhesion to host surfaces, are suitable criteria for selection of candidate probiotics for use in the culture of Senegalese sole.     

Interference of Listonella anguillarum with potential probiotic microorganisms isolated from farmed gilthead seabream (Sparus aurata, L.)

Four isolates obtained from gilthead seabream have been tested for their adhesion to the skin, gill and intestinal mucus of gilthead seabream, and for their ability to interfere with Listonella anguillarum, an important pathogen of farmed gilthead seabream. The ability to adhere to mucus was higher than 7% for all isolates. Three isolates showed an antagonistic effect against some of the pathogenic strains tested. They were assayed to interfere with the attachment of L. anguillarum to the mucus of gilthead seabream. Only two isolates significantly reduced the adhesion of L. anguillarum to all of the mucus assayed under exclusion, competition and displacement conditions. According to the criteria applied, the isolate Pdp11 was selected and its in vivo probiotic potential was assessed by oral administration followed by challenge with the pathogen L. anguillarum. For the feeding trial, a group of 50 gilthead seabreams received a commercial diet supplemented with lyophilized 10⁸ CFU g^?1 of this isolate for 15 days. An other group of similar characteristics received the non‐supplemented commercial diet. After the challenge, the mortality of the fish receiving the diet supplemented with the potential probiotic was significantly lower than that observed in the groups of fish receiving the non‐supplemented commercial diet.     

Is the turbot,Scophthalmus maximus (L.), intestine a portal of entry for the fish pathogen Vibrio anguillarum?

The role of the intestinal tract in Vibrio anguillarum infection of turbot, Scophthalmus maximus (L.), fingerlings was investigated in two in vivo models and the possible mechanisms involved were studied in vitro. Viable V. anguillarum cells were detected in spleens from more than 50% of the fish administered the pathogen orally or rectally, suggesting that the intestinal tract is a portal of entry for V anguillarum. In transmission electron micrographs, V. anguillarum- like cells were seen close to the rectal epithelium, suggesting penetration of the mucus layer, but no epithelial cell penetration or endocytosis was evident. Attachment to intact turbot intestines was investigated, and 80% or more of the bacterial cells still remained attached after serial washings. A significantly higher number of cells attached to rectal segments than to the other intestinal segments. In vitro, V. anguillarum cells did not adhere specifically to intestinal mucus, but rather accumulated close to intestinal mucus interfaces and subsequently penetrated them. It is proposed that the intestinal ttact of tutbot is a portal of entry for V. anguillarum and that the cells penetrate the intestinal mucus overlaying the epithelial cells.     

Evaluating the probiotic potential and adhesion characteristics of Bacillus spp. isolated from the intestine of Rhynchocypris lagowskii Dybowski

Probiotics have been widely used in aquaculture worldwide especially in China as antibiotics have been banned in aquaculture. In the present study, 27 potential strains were isolated from Rhynchocypris lagowskii Dybowski intestinal tract towards the assessment of their probiotic potential. Two potential probiotics were finally screened from the 27 candidate strains according to the results of enzyme-producing ability, in vitro against pathogens ability and antibiotic sensitivity test. The 2 potential probiotics were identified and confirmed on the basis of their colony morphology, biochemical characteristics, and 16S rRNA sequencing. The probiotic strains, LSG2-3–2 and LSG3-6, were identified to be Bacillus methylotrophicus and B. tequilensis, respectively. Further studies showed that LSG2-3–2 and LSG3-6 had excellent tolerance to high temperature (80℃), low pH (3–5), bile salts (0.3%), intestinal juice (10%), and gastric juice (0.5%). The adhesion rates of LSG2-3–2 and LSG3-6 in the in vitro intestinal mucosal adhesion model were 17.74% and 24.04%, respectively. Analysis of their bacteria surface adhesive proteins revealed that the lectins on the bacterial surface of LSG2-3–2 and LSG3-6 were mainly protein and glycoprotein properties, respectively. The adhesion receptor components in the mucus proteins of the two strains were all protein properties. The results of the inhibitory adhesion test indicated that LSG3-6 had a higher inhibitory effect on Aeromonas hydrophila and LSG2-3–2 had a better inhibitory effect on A. veronii. The biosafety assay confirmed that the isolates were not pathogenic to the host fish. Based on the presently observed characteristic features, it can be concluded that LSG2-3–2 (B. methylotrophicus) and LSG3-6 (B. tequilensis) retrieved from the digestive tract of R. lagowskii can be used for the healthy breeding and disease prevention of R. lagowskii.

     

In vitro selection and characterization of putative probiotics isolated from the gut of Acipenser baerii (Brandt, 1869)

To select and characterize potential probiotic bacteria from the gut microbiota of Siberian sturgeon (Acipenser baerii), 129 strains isolated from the hindgut were screened for antagonistic activity against five fish pathogens. Ten isolates showed antagonism towards three or more pathogens. Nine of these isolates were Gram‐positive, belonging to Lactococcus (seven) and Bacillus (two), and a single strain belonging to the Gram‐negative Citrobacter. These inhibitory isolates were identified using genetic, phentotypic and biochemical traits, and further characterized by in vitro tests assessing the adhesion and growth in mucus and resistance to gastric and intestinal fluids. The candidate probiotics were determined to be non‐pathogenic through an in vivo study. Based on these assays, Lactococcus lactis ssp. lactis STG45 and STG81 showed the broadest inhibitory potential, a high viability in simulated gastrointestinal juice and the highest adhesion capacity to mucus. They were therefore selected as the most promising candidate probiotics. This is the first study screening probiotics among the gut microflora of Siberian sturgeon.     

Chemotaxis towards, adhesion to, and growth in carp gut mucus of two Aeromonas hydrophila strains with different pathogenicity for common carp, Cyprinus carpio L

Characteristics that promote bacterial colonization of the intestinal mucosal surface were examined in two strains of the common fish pathogen Aeromonas hydrophila, with different pathogenicity. The characteristics examined were chemotactic activity towards mucus, bacterial adherence to mucus and growth in mucus. Intestinal gut mucus of healthy common carp was used. The results indicate that chemotaxis is not necessary for a bacterium to become pathogenic, but it may be a necessary parameter for a bacterium to be an obligate pathogen. Adhesion also seems to be a factor influencing pathogenicity. The results suggest that higher adhesion to mucus and subsequent growth is associated with differences in pathogenicity.     

Aqualase®, a yeast‐based in‐feed probiotic,modulates intestinal microbiota,immunity and growth of rainbow trout Oncorhynchus mykiss

Yeast probiotics have great promise, yet they received little attention in fish. This study investigated the influence of Aqualase^®, a yeast‐based commercial probiotic composed of Saccharomyces cerevisiae and Saccharomyces elipsoedas, on health and performance of rainbow trout (Oncorhynchus mykiss). Probiotics were incorporated in the diets at three different inclusion levels (1%, 1.5% and 2%) and administered to the fish for a period of 8 weeks. After the feeding trial, intestinal total viable aerobic bacterial count was significantly higher in fish group that received 2% in‐feed probiotics. In addition, a significant increase in at least 11% in intestinal lactic acid bacteria population was observed in all probiotic‐fed groups. Total protein level and lysozyme activity in skin mucus were significantly elevated following probiotic feeding. Inhibitory potential of skin mucus against fish pathogens was significantly enhanced by at least 50% in probiotic‐fed groups. Humoral and cellular immune parameters were influenced by probiotic feeding and the effects were dependent on inclusion level. Digestive physiology was affected by in‐feed probiotics through improvement of intestinal enzyme activities. All growth performance parameters were significantly improved following probiotic administration specifically at inclusion rate 1.5% and above. Taken together, the results revealed that Aqualase^® is a promising yeast‐based probiotic for rainbow trout with the capability of modulating the intestinal microbiota, immunity and growth.     

鲫肠道乳酸菌的分离及益生特性

为获得鱼源益生乳酸菌,本研究从健康鲫肠道内分离鉴定得到38株乳酸菌,并选择其中8株乳酸菌进行体外益生特性分析。结果显示,8株乳酸菌均能在p H=4.5和10%鲫胆汁环境中存活,对嗜水气单胞菌、豚鼠气单胞菌、无乳链球菌和副溶血弧菌均有较强的抑菌能力,但菌体表面疏水性和自凝集能力具有菌株特异性。选择5株疏水性较高(63%~89%)、自凝集能力强(37%~45%)的乳酸乳球菌进行体外黏附能力测定,结果显示,黏附能力在菌株间具有差异性(4.5%~7.9%),但均能显著降低嗜水气单胞菌NJ-35的体外黏附率,黏附抑制率达30%~35%;最后对筛选出的3株高黏附力的乳酸乳球菌进行安全性评价,发现3株菌对鲫均无致病力。鱼源乳酸菌的筛选,为鲫养殖生产中益生菌的实际应用提供了理论依据。     

解淀粉芽孢杆菌和胶红酵母复合菌对虹鳟生长性能及胃黏膜、肠黏膜菌群结构的影响

为研究饲料中添加不同水平的复合菌剂(解淀粉芽孢杆菌,Bacillus amyloliquefaciens V4和胶红酵母,Rhodotorula mucilaginosa)对虹鳟(Oncorhynchus mykiss)幼鱼生长及消化道黏膜微生物菌群结构的影响,选用体重为(205.1±4.82)g的虹鳟幼鱼360尾,随机分为4组(每组3个重复,每个重复30尾),分别投喂基础饲料(C0)和3种添加水平为5×10~6/5×10~7 CFU/g(T1),1.5×10~7/1.5×10~8 CFU/g(T3),2.5×10~7/2.5×10~8 CFU/g(T5)的复合菌剂(B.amyloliquefaciens V4/R.mucilaginosa),实验周期42 d。研究结果发现饲料中添加复合益生菌对虹鳟的生长及存活有一定的促进和提高,T1比例的复合益生菌能够显著提高虹鳟的增重率和特定生长率、显著降低饲料系数(P0.05),同时T1和T3比例添加显著降低虹鳟的死亡率(P0.05);对其消化道黏膜细菌群落16S rDNA进行聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)指纹分析,结果表明:虹鳟胃黏膜和肠黏膜上微生物菌群种类存在差异;胃黏膜菌群DGGE图谱中分别检测到35.7±17.0(C0)、37.0±3.5(T1)、36.7±13.6(T3)、26.0±13.2(T5)条谱带,各处理组间谱带数目无显著差异(F=0.500,P=0.692),肠黏膜菌群DGGE图谱显示分别检测到23.3±5.8(C0)、22.3±3.2(T1)、16.7±8.0(T3)、24.7±7.4(T5)条谱带,各处理组间谱带数目也无显著差异(F=0.916,P=0.475);胃黏膜菌群多样性随着益生菌添加量增加,菌群多样性有升高趋势,但是在最高浓度组(T5)多样性降低,肠黏膜菌群多样性随着复合菌剂的添加,多样性指数持续降低,中浓度添加组(T3)多样性最低,但是随着添加浓度升高,呈现恢复和升高趋势(T5);基于所得PCR-DGGE指纹图谱中谱带丰度值数据的UPGMA聚类和PCA排序分析均显示胃黏膜微生物群落与肠黏膜微生物群落结构差异明显,大致分为两个不同的分支,胃黏膜和肠黏膜微生物菌群并没有按照不同处理组而有显著分化。以上结果表明解淀粉芽孢杆菌和胶红酵母复合添加能显著促进虹鳟的生长,提高存活率,外源益生菌添加对虹鳟消化道黏膜上优势菌群能产生一定影响,但并未对胃黏膜及肠黏膜菌群多样性产生显著影响,同时也未显著改变虹鳟肠黏膜微生物菌群结构,高比例添加降低消化道黏膜细菌数量及多样性风险。     

Kinetics of adhesion of Listonella anguillarum to the mucus of gilt-head seabream, and the implication of surface components

The kinetics of adhesion of Listonella anguillarum cells to skin, intestinal and gill mucus suspensions of gilt‐head seabream was studied. A Langmuir adsorption isotherm was calculated, and the results obtained indicated that this microorganism showed a saturation kinetics. The pretreatment of the mucus suspensions with a surface extract obtained from L. anguillarum cells promoted a very important inhibition of the number of adhered bacteria to the different mucus suspensions. The potential role of surface structures on the adhesion of L. anguillarum to mucus of gilt‐head seabream is discussed.