Pseudomonas putida infection in rainbow trout

A disease outbreak occurred during the Spring of 2006 among rainbow trout, Oncorhynchus mykiss, kept in growout tanks. During the outbreak 35% of the rainbow trout died. Typical clinical signs were ulceration on the dorsal surface of the fish. Based on biochemical characteristics and sequence of 16S rRNA, the causative bacteria isolated from diseased fish were identified as Pseudomonas putida. Histologically, skin ulcer initially was characterized by sloughed off epithelia and epithelial necrosis. In the advanced form of the disease, almost all layers of the skin down to the epidermis were lost. Outside of Japan, this study reports the first P. putida infection in fish.     

Disease of rainbow trout caused by Pseudomonas luteola

Bacteria isolated from rainbow trout, Oncorhynchus mykiss, kept in a farm, in Turkey. During the outbreak, 40% of the rainbow trout (10-40 g) died. Typical clinical signs were exophthalmia, dark pigmentation, hemorrhage at the base of the pectoral, pelvic, anal fins and around the vent. Internal signs were enlarged spleen, pale liver and intestine filled with yellowish fluid. Liver, kidney and spleen of diseased fish, were aseptically streaked on Tryptic Soy Agar. After incubation, pure cultured colonies were observed and biochemically characterized with API 20 NE and other biochemical tests. Cultured bacterial 16 S rDNA gene was sequenced. Based on biochemical characteristics and sequence of 16 S rRNA, the causative bacteria were identified as Pseudomonas luteola. This study reports the first P. luteola infection in fish.     

Exploring the microbial diversity of the distal intestinal lumen and mucosa of farmed rainbow trout Oncorhynchus mykiss (Walbaum) using next generation sequencing (NGS)

In this study, next generation sequencing (NGS) was used to survey the 16S rRNA ribotypes of the distal intestinal lumen and mucosal epithelium of farmed rainbow trout. This approach yielded a library consisting of 2 979 715 quality filtered paired sequences, assigned to genus level of taxonomy using the Ribosomal Database Project (RDP). A high level of diversity was observed in both regions. A total of 90 bacterial genera were identified in the lumen of all fish sampled, compared with 159 in the mucosa. The allochthonous microflora was dominated by sequences belonging to the γ Proteobacteria (mean sequence abundance 54.3%), in particular the Enterobacteriaceae, with Yersinia, Serratia, Hafnia and Obesumbacterium the most abundant genera. Fewer γ Proteobacteria (mean sequence abundance 37%) were present in the mucosa, and autochthonous communities consisted of a more even split among the bacterial classes, with increases in sequences assigned to members of the β Proteobacteria (mean sequence abundance 18.4%) and Bacilli (mean sequence abundance 16.8%). The principal bacterial genera recorded in the mucosa were Cetobacterium, Yersinia, Ralstonia, Hafnia and Carnobacterium. The results of this study demonstrate that the luminal and mucosal bacterial communities may be different in their respective structures, and that the mucosal microflora of rainbow trout may be more diverse than previous research has suggested. This research also demonstrated a degree of conservation of bacterial genera between individual fish sampled, and is to the author's knowledge the first time the MiSeq^® NGS platform has been used to explore the rainbow trout intestinal microflora.     

摄食不同饵料的大口黑鲈肠道菌群分析

为了探究投喂冰鲜杂鱼和人工配合饲料对大口黑鲈肠道菌群的影响,本研究采用变性梯度凝胶电泳技术(PCR-DGGE)对摄食不同饵料的2组鲈鱼肠道内容物的细菌群落结构进行了比较分析.结果显示,冰鲜组与饲料组的大口黑鲈肠道菌群组成相似性较低(相似性24％),且冰鲜组肠道菌群多样性高于饲料组.对2组大口黑鲈中共有条带和特异性条带进一步测序分析显示,2组大口黑鲈肠道共有菌有红螺菌、梭菌、保科爱德华菌、嗜麦芽窄食单胞菌和蓝细菌.冰鲜组大口黑鲈肠道中的特异性菌归属为拟杆菌、肉食杆菌、魏斯氏菌和邻单胞菌;饲料组大口黑鲈肠道中的特异性菌归属为黄杆菌、鲸杆菌、柠檬明串珠菌、缺陷短波单胞菌.研究表明,与投喂冰鲜杂鱼的大口黑鲈相比,人工配合饲料降低了大口黑鲈肠道菌群多样性,也抑制了拟杆菌等有益菌在大口黑鲈肠道内的分布.推测饲料组大口黑鲈生长较慢可能与其肠道菌群中拟杆菌与厚壁菌的比值下降有关.研究结果提示,在今后大口黑鲈养殖或者人工配合饲料研制过程中,添加拟杆菌等益生菌也许可促进大口黑鲈的生长和营养吸收.     

Effects of soybean meal based diet on growth performance, gut histopathology and intestinal microbiota of juvenile rainbow trout (Oncorhynchus mykiss)

Effects of soybean meal (SBM) based diet on growth performance, histology of the intestinal epithelium and on the gut microbiota of juvenile rainbow trout (Oncorhynchus mykiss) were investigated on two trials lasting for 8 weeks (Trial I) and 18 weeks (Trial II). The microbiological characterization was done both with conventional plating techniques, biochemical profiling and length heterogeneity analysis of PCR amplified 16S rDNA (LH-PCR).Typical histological changes were found after 18 weeks of SBM feeding (Trial II). Mean height of simple foldings were significantly higher within the group fed with fish meal (FM) based diet. These changes were not, however, reflected in the appearance, behavior and weight gain of the fish, although the feed conversion rate (FCR) was higher, compared to fish fed with the traditional FM feed.In both SBM and FM groups the counts of cultivable intestinal bacteria dropped within the first 4 weeks of feeding (Trial I). Subsequently, the bacterial numbers increased in the FM group, but not in the SBM group by the 8th week of the trial. The LH-PCR data suggested also that diet-related qualitative changes in the microbiota had occurred (Trial II). The main identified bacteria were mainly representatives of the genera Aeromonas, Sphingomonas and Chryseomonas, and among the lactic acid bacteria, Lactococcus and Lactobacillus.Future research will be directed to the identification and eventual elimination the SBM-associated factors involved in the inflammatory responses and unfavorable FCR, in particular by modifying the intestinal microbiota.     

Effect of Thymus vulgaris essential oil on intestinal bacterial microbiota of rainbow trout,Oncorhynchus mykiss (Walbaum) and bacterial isolates

The application of natural and innocuous compounds has potential in aquaculture as an alternative to antibiotics. We evaluated the effect of diet supplementation with Thymus vulgaris essential oil (TVEO) on the allochthonous microbial composition of rainbow trout. DNA was extracted directly from the intestinal contents, and the V3‐V4 regions of the 16S rRNA genes were amplified by PCR. The bacterial composition was analysed using temporal temperature gradient electrophoresis (TTGE). No significant changes (P>0.05) were detected in the TTGE profiles of TVEO‐treated trout compared with the controls. The Dice similarity index revealed a high stability (C_s >70%) of the intestinal microbiota in both groups during the 5‐week period. Sequence analyses of the TTGE bands revealed the same bacterial composition in both groups, with most bacteria belonging to the Proteobacteria and Firmicutes phyla. The in vitro antibacterial activity of TVEO was assessed using a range of normal intestinal isolates and fish pathogens. The inhibitory concentrations for all the tested bacteria were higher than the TVEO levels used in trout, which may explain the in vivo results.     

Recovery of Bacillus mycoides,B. pseudomycoides and Aeromonas hydrophila from common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) with gill disease

During a 3‐month period from June to the end of August 2016, ~5% mortalities were observed in a farm with rainbow trout (Oncorhynchus mykiss Walbaum) and one farm of common carp (Cyprinus carpio L.) in Bulgaria. The disease was manifested by gill ulcers/rot, asphyxiation and bloody ascites. Aeromonas hydrophila was isolated from the internal organs of all the diseased fish. Bacillus mycoides or B. pseudomycoides were recovered from the gill lesions on diseased carp and rainbow trout, respectively, with identification achieved by conventional phenotyping and by sequencing of the 16S rRNA gene. In vivo experiments confirmed that all three organisms were pathogenic to rainbow trout.     

A comparative molecular study of the presence of “Candidatus arthromitus” in the digestive system of rainbow trout,Oncorhynchus mykiss (Walbaum), healthy and affected with rainbow trout gastroenteritis

Observations were made using histopathological techniques in conjunction with a nested polymerase chain reaction (PCR) protocol for the specific detection of “Candidatus arthromitus” on DNA extracted from wax‐embedded tissues and fresh digestive contents of rainbow trout. Samples positive for “Candidatus arthromitus” DNA included fish with rainbow trout gastroenteritis (RTGE), clinically normal cohabiting fish, and apparently healthy controls from RTGE positive and RTGE negative sites. The results obtained from the PCR were confirmed by nucleotide sequencing. “Candidatus arthromitus” DNA was found in distal intestine as well as in sections of pyloric caeca, suggesting that both these locations are appropriate for molecular detection of “Candidatus arthromitus” DNA in trout. Furthermore, rainbow trout fry distal intestinal samples from two different hatcheries where RTGE had not been reported were also positive. Differences in “Candidatus arthromitus” DNA detection between paraffin wax‐embedded and fresh digestive content samples from the same fish suggested that it may be predominantly epithelium‐associated in healthy trout. Parallel histopathological observations indicated that pyloric caeca are the preferred site for visualizing segmented filamentous bacteria (SFB) in trout with RTGE. The results of this study showed that the presence of SFB was not invariably associated with clinical disease and that more information is required to understand the role of these organisms.     

The growth of rainbow trout (Salmo gairdneri) given diets containing chitin and its relationship to chitinolytic enzymes and chitin digestibility

The growth of rainbow trout fed diets containing 4, 10 and 25% chitin, over a 12-week period, was significantly depressed (P < 0.001) when compared with controls fed diets containing 25% starch. There was no difference in growth rate between control fish and those fed diets containing 10% N-acetyl glucosamine (GlcNAc). Isotopically labelled amino sugars were shown to be oxidized when injected intraperitoneally into rainbow trout. Relatively high levels were found of chitinase activity in the stomachs and of chitobiase in the intestines. These enzyme activities were similar in all the trout, irrespective of the amount of chitin in their diets, except that chitobiase in the intestines of fish fed diets containing GlcNAc showed higher levels of activity than the controls (P < 0.05). Chitin was not significantly digested when fed at 10 and 30% of the diet but the apparent digestibility of pre-cooked starch was 50% when fed at either 15 or 25%, on the basis of the inert indicator (Cr₂O₃) method. Chitinase and chitobiase activities were not reduced in fish fed diets containing 10% chitin and an antibiotic (Tribrissen^®) although the alimentary microflora were completely eliminated. Enzyme activities were not enhanced when live chitinolytic bacteria (Vibrio alginolyticus) were incorporated into diets with 10% chitin; these bacteria were only recovered from the intestine. The evidence indicates an endogenous origin of chitinolytic enzymes in the trout gastro-intestinal tract. The presence of either antibiotic or bacteria in the diet had no effect on the digestibility of chitin.     

Effects of low‐level dietary microalgae supplementation on the distal intestinal microbiome of farmed rainbow trout Oncorhynchus mykiss (Walbaum)

In this study, high throughput 16S rRNA sequencing was used to investigate the effect of a novel whole‐cell dietary microalgae meal (Schizochytrium limacinum), on the distal intestinal microbiome of farmed rainbow trout, Oncorhynchus mykiss. Heterotrophic microalgae are rich in omega 3 polyunsaturated fatty acids, can be produced sustainably and have been shown to have beneficial effects on host health. After a 15‐week trial period, microbial community profiles were compared between the distal intestinal contents of fish fed either a control diet or a treatment diet that partially replaced fish oil with microalgae meal, at a substitution level of 5%. The results of this research showed that the microbial communities of both fish populations were composed of similar microbial taxa, however, the treatment group fed the microalgae supplement possessed a greater level of microbial diversity than those in the control group. A limited number of bacterial taxa were discriminatory between diets and were significantly elevated in the treatment group, notably operational taxonomic units (OTUs) assigned to the genera Streptococcus, Leuconostoc, Lactobacillus, Lactococcus and Weissella. However, the overall structure of the intestinal microbiome between control and treatment groups was not found to be significantly different. The treatment group displayed a heavier mean weight and condition factor at the end of the trial period. The results of this study suggest that the tested microalgae meal can be used as a replacement for a proportion of fish oil in aquafeeds, with minor changes to the intestinal microbiome of farmed rainbow trout, and positive effects on growth.     

Dietary probiotic Pediococcus acidilactici MA18/5M modulates the intestinal microbiota and stimulates intestinal immunity in rainbow trout (Oncorhynchus mykiss)

A study was conducted to evaluate the probiotic effect of Pediococcus acidilactici MA18/5M on rainbow trout, Oncorhynchus mykiss. Fish (310 ± 9 g) were fed a control diet or a P. acidilactici‐supplemented diet (at 2.4 × 10⁶ CFU/g) for 4 weeks. The probiotic was observed to populate the intestine with levels ranging from log 3.7 to 5.4 CFU/g. Furthermore, these populations were able to persist for at least 24 hr after the cessation of probiotic feeding. High‐throughput sequencing analysis of bacterial 16S rRNA libraries demonstrated that P. acidilactici was able to modulate the gut microbiome of rainbow trout and that the probiotic was detected as a common taxon on the mucosa and in the digesta of the probiotic fish (p < .05). Real‐time polymerase chain reaction demonstrated that feeding the probiotic upregulated pro‐inflammatory cytokines, interleukin‐1β, and interleukin‐8 and downregulated anti‐inflammatory interleukin‐10 compared to the control‐fed fish. Furthermore, the mRNA levels for the mucosal antibody immunoglobulin T was also elevated in probiotic‐fed fish. These findings help to explain some of the mechanisms behind the previously reported observed benefits of using this probiotic in the intestinal morphology and immunity of rainbow trout.     

Clinical and aetiological aspects of a summer enteritic syndrome associated with the sporulating segmented filamentous bacterium 'Candidatus Arthromitus' in farmed rainbow trout, Oncorhynchus mykiss (Walbaum)

An enteritic syndrome affecting farmed rainbow trout and occurring predominantly in summer has recently been described and related to the massive accumulation of a segmented filamentous bacterium (SFB), ‘Candidatus Arthromitus’, in the digestive tract of fish. Details of the clinical manifestations, the characteristics of the bacterium and the possible aetiology of the disease are presented in this paper. No other bacterial pathogen than ‘Candidatus Arthromitus’ was characterized in trout undergoing the enteritic syndrome. From clinical and histopathological observations, a toxin or toxic product accumulation was suspected in affected trout, and could account for the massive destruction of the intestinal mucosa. However, the origin and aetiological role of the SFB could not be clearly elucidated, although most of the filaments appeared to beviable and to produce and release strings of endospores in the lumen of the gut. The organism could not be cultured in the conditions tested, shared several morphological and structural characteristics with related bacteria described in the intestinal microflora of different vertebrate species, but no attachment structure was definitively recognized. All attempts to detect similar SFB in the fish diet orin the environment of affected fish farms were unsuccessful. It is unclear whether this organism represents a normal and not previously detected inhabitant of the intestine of trout stressed after environmental perturbation, or whether it results directly from external sources of contamination.     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

Effects of biotechnologically processed soybean meals in a nonfishmeal diet on growth performance, bile acid status, and morphological condition of the distal intestine and liver of rainbow trout Oncorhynchus mykiss

This study evaluated four commercially available biotechnologically processed soybean meals (SBMs) that were included as the primary protein sources in a nonfishmeal diet for juvenile rainbow trout Oncorhynchus mykiss. The processed SBMs were Afcep SBM fermented with mixed bacteria predominantly Bacillus spp., DaBomb SBM fermented with Lactobacillus acidophilus, PepSoyGen SBM fermented with Aspergillus oryzae and Bacillus subtilis, and HP300 SBM treated enzymatically. Fishmeal (FM) and common heat-treated and hulled SBM were used in the reference diets. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) analysis revealed that the protein components of Afcep SBM appeared to have relatively small molecular sizes compared with the other SBM products. After a 10-week feeding trial, growth of fish fed the common SBM diet was depressed, and abnormal liver and distal intestinal morphologies and bile acid characteristics were observed. The Afcep SBM diet showed approximated growth performance and comparable physiological conditions relative to the FM diet, while only limited improvements were observed in fish fed the other processed SBMs. These findings suggest that Afcep SBM, with its smaller molecular mass proteins, is the most promising processed SBM in nonfishmeal diets for rainbow trout.     

Phylogenetic analysis of intestinal bacteria of freshwater salmon Salmo salar and sea trout Salmo trutta trutta and diet

The microbial diversity of culturable intestinal microflora of wild freshwater salmonid fishes salmon Salmo salar and sea trout Salmo trutta trutta juveniles (0+ years old) from the same environmental conditions were investigated by means of molecular identification techniques and analysis of diet. Significant differences in the intestinal microbial diversity were observed in different fish species. The predominant group in the intestinal tract of the salmon comprised representatives of the Enterobacteriaceae family (23%), Plesiomonas (19.2%) and Carnobacterium (15.3%). Predominant microbiota in sea trout intestinal tract were Enterobacteriaceae (52%), Aeromonas (22%) and Pseudomonas (14%). The results show that Enterobacteriaceae were predominant in the intestinal tract of the salmon and the sea trout juveniles raised on diets of different compositions. However, molecular identification of the intestinal microbiota at the species or genus level revealed differences in these fish species. Bacteria in the hindgut of salmon included Pragia and Serratia. However, bacteria in the gut contents of sea trout from the Enterobacteriaceae family were Buttiauxella, Enterobacter, Moellerella, Pantoea, Rahnella and Tiedjeia arctica. A novel phylotype of T. arctica is harbored in the intestinal tract of wild salmon, and may correspond to a previously undescribed species.     

Influence of dietary soy protein and peptide products on bile acid status and distal intestinal morphology of rainbow trout Oncorhynchus mykiss

To investigate the influence of dietary soy protein size on the bile acid status and the distal intestinal morphology in rainbow trout Oncorhynchus mykiss, diets based on soybean meal (SBM), soy protein isolate (SPI) and 4 soy peptide products having different molecular sizes were fed to trout (initial BW, 19?g) for 10?weeks. A diet based on fish meal (Cont) and a diet including the smallest-peptide and low-saponin product (Hi-Nute AM) and supplemented with soya saponin (AMS) were also fed. In fish fed diets SBM and SPI, the gallbladder was atrophied, the proportion of biliary cholyltaurine decreased, and highly vacuolated epithelial cells of mucosal folds and proliferation of connective tissues in the submucosa of the distal intestine were observed. These parameters tended to improve in fish fed soy peptide diets and especially those of fish fed diet AM were similar to fish fed diet Cont. Although the gallbladder of fish fed diet AMS was smaller than fish fed the unsupplemented diet AM, no morphological abnormalities were observed in the distal intestine. These results suggest that soy proteins/peptides with relatively large sizes and with soya saponin affect the bile acid status and distal intestinal morphology of rainbow trout.     

两种疾病发生对养殖半滑舌鳎肠道菌群结构的影响分析

为研究疾病发生情况下鱼类肠道菌群的结构变化,运用高通量测序方法分别对感染腹水病和皮肤溃疡病的工厂化养殖半滑舌鳎病鱼及同一养殖系统中健康鱼的肠道菌群结构进行了对比分析。结果发现,养殖半滑舌鳎肠道中的细菌种类要明显少于环境中的细菌种类,仅为养殖池进水和颗粒饲料中细菌种类总数的14.3%~44.7%。在疾病发生后,病鱼肠道中的细菌种类数量仅是同一养殖系统中健康鱼肠道细菌种类数量的45.8%和64.5%,病鱼肠道中细菌多样性下降非常明显。进一步分析肠道菌群结构的组成发现,虽然发病鱼和健康鱼肠道中超过60%的细菌种类不相同,但优势菌群的结构组成差异不明显,丰度最高的前10种细菌种类基本相同,其比例超过肠道相对总菌量的80%。不同疾病的发生对半滑舌鳎肠道菌群结构的影响也不同,腹水病会造成肠道中优势菌株的种类和相对丰度都发生比较明显的改变,而皮肤溃疡病发生前后肠道菌群结构组成几乎没有变化。研究表明,养殖半滑舌鳎的肠道菌群结构相对比较保守,而且并不是所有的疾病种类都会引起肠道菌群结构发生改变。     

Detection of the bacterium Flavobacterium psychrophilum from a natural infection in rainbow trout, Oncorhynchus mykiss (Walbaum), using formalin-fixed, wax-embedded fish tissues

The bacterial pathogen Flavobacterium psychrophilum was successfully identified from formalin-fixed, wax-embedded tissue blocks of infected rainbow trout heart and spleen tissues, using a polymerase chain reaction (PCR)-based assay. Filamentous bacteria were observed in haematoxylin and eosin and Giemsa-stained sections but no bacteria were recovered from the diseased fish using standard bacteriology isolation techniques. All infected fish had histopathological evidence of myocarditis or rainbow trout fry syndrome. Immunohistochemistry was attempted using three different anti-F. psychrophilum sera but the results were inconclusive, and an alternative molecular approach was therefore attempted. This paper describes the use of a PCR-based assay to help identify bacteria present in formalin-fixed, wax-embedded tissue samples. This is the first time that this technique has been used for the detection of fish bacteria from diagnostic samples.     

Identification and characterization of lactic acid bacteria isolated from rainbow trout (Oncorhynchus mykiss,Walbaum 1792), with inhibitory activity against Vagococcus salmoninarum and Lactococcus garvieae

In this study, a total of 98 lactic acid bacteria isolated from rainbow trout intestines were screened for their probiotic properties. The isolates were tested for their ability to inhibit growth of Vagococcus salmoninarum and Lactococcus garvieae. Based on in vitro antagonism, 10 isolates were selected and evaluated pathogenicity in rainbow trout. Isolates were further investigated for hydrophobicity, bile salts and acid tolerance. These isolates were able to survive low pH and high bile concentrations and showed good adherence characteristics. Isolates were characterized phenotypically, and then, 16S rRNA gene sequence analysis was used for confirmation. Selected strains were administered orally at 10⁸ cfu/g feed, and fish were challenged with V. salmoninarum and L. garvieae. The fish fed with lactic acid bacteria supplemented diets did not improve protection against V. salmoninarum. However, administration of Lactococcus lactis subsp. lactis M17 2‐2 and Lactobacillus sakei 2‐3 resulted in a significant reduction in mortality due to L. garvieae when compared to the control fish. RPS values were calculated as 80 and 53% in fish fed with L. sakei 2‐3 and L. lactis subsp. lactis M17 2‐2, respectively. Our results suggest that these strains could provide an alternative for lactococcosis control in aquaculture.     

Influence of soya saponin, soya lectin, and cholyltaurine supplemented to a casein-based semipurified diet on intestinal morphology and biliary bile status in fingerling rainbow trout Oncorhynchus mykiss

To investigate the cause of the changes in intestinal morphology and biliary bile status of rainbow trout Oncorhynchus mykiss fed defatted soybean meal (SBM)-based diets, casein-based semipurified diets supplemented with soya saponin, soya lectin, and cholyltaurine were fed to rainbow trout for 6 weeks. An unsupplemented control diet and a SBM-based diet were also tested as references. Poor development of microvilli and pinocytotic vacuoles, and accumulation of large vacuoles in the epithelial cells were observed in the distal intestine of fish fed diets containing saponin but not cholyltaurine. Hyperplastic connective tissue in the mucosal folds of the distal intestine was observed in fish fed a diet containing both saponin and lectin but not cholyltaurine. However, intestinal histological features in fish fed diet supplemented with cholyltaurine and lectin and/or saponin were similar to those in the control diet group. Liver morphology and biliary bile status were not affected by saponin and lectin. These results suggest that the abnormal features of the distal intestine of rainbow trout fed SBM-based diets are caused by the combination of soya saponin and soya lectin, and that supplemental cholyltaurine plays certain roles in normalizing the intestinal abnormalities caused by the saponin and lectin.