Immunogenicity and protective efficacy of Yersinia ruckeri lipopolysaccharide (LPS), encapsulated by alginate-chitosan micro/nanoparticles in rainbow trout (Oncorhyncus mykiss).

Considering the many advantages of oral vaccines in aquaculture, several studies have been conducted in this area recently. In this study, immunization and protective power of the oral vaccine of Yersinia ruckeri encapsulated with Alginate-Chitosan micro/nanoparticles were evaluated in rainbow trout. For this purpose, 720 juvenile rainbow trout (9 ± 1.8 g) were divided into 8 groups in three replications (30 fish each) as follows: Groups A, B and C, were immunized with Yersinia ruckeri lipopolysaccharide (LPS), LPS+Formalin Killed Cells (FKC) and FKC alone, groups D, E, and F were immunized with encapsulated LPS, LPS+FKC and FKC, respectively. The G and H groups considered as encapsulated and non-encapsulated control, respectively. Micro/nanoencapsulation with alginate-chitosan was performed by internal emulsification method and vaccination were conductrd in the first and third weeks via oral route. Sampling was performed on days 0, 30, and 60 of experiment. Anti Y. ruckeri antibody titer in serum, intestine and skin mucus were measured via ELISA method. Non-specific immune response including: serum lysozyme, complement, bactericidal and respiratory burst activity, serum protein and globulin level, as well as white blood cell count were compared among the groups. The expression of IgT gene in the intestine and TCR gene in the anterior kidney were also investigated. At the end of the study, the fish were challenged with Y. ruckeri through immerssion and intraperitoneal routs and the relative survival rate was evaluated. Result showed that the antibody level in serum, skin and intestine was significantly higher in group E and F than control groups (P < 0.05), meanwhile serum, skin and intestine antibody level in all vaccinated groups were significantly (P < 0.01) higher in day 30 and 60 compare to zero day. Non-specific immunity factors including: serum lysozyme, complement, and respiratory burst activity as well as WBC, protein and Globulin level were significantly higher in E and F groups not only in day 30 but also in day 60 of experiment (P < 0.05). Cumulative mortality following injection and bath challenge were significantly (P = 0.004) lower (35%-45%) in groups E and F compare to control group (80%). The IgT and TCR gene expression in groups D, E and F were significantly higher (P < 0.05) than control group. Highest upregulation of IgT and TCR gene expression in vaccinated groups were seen at day 30 and 60 respectively which were significantly (P < 0.001) higher than day zero. Generally, it can be concluded that nano/micronanoencapsulation of Y. ruckeri FKC+LPS with chitosan-alginate, not only increases protective efficacy of oral vaccine, but improves specific and non-specific immune responses in rainbow trout.

Effect of Encapsulated Lactobacillus bulgaricus on Innate Immune System and Hematological Parameters in Rainbow Trout (Oncorhynchus mykiss), Post-Administration of Pb.

The present study was conducted to investigate the effects of probiotic and encapsulated Lactobacillus bulgaricus on hematological and immunological factors after lead toxicity in rainbow trout (Oncorhynchus mykiss). Two hundred and forty fish weighing about 16 ± 3.8 g were divided randomly in to four groups including two groups which were fed by a diet containing ~ 108 CFU g-1Lactobacillus bulgaricus and encapsulated Lactobacillus bulgaricus bacteria and also the third group diet without Lactobacillus bulgaricus. After 45 days, in addition to probiotic (~ 108 CFU g-1), 500 µg kg of lead nitrate was added to the food of the three groups for 21 days. The fourth group (control) was first fed to the normal diet for 45 days then exposed to Pb. Blood samples were collected at days 45, 52, 59, and 66, and hematological and some immunological parameters were assessed. Results showed that hemoglobin, red blood cells, white blood cells, and lysozyme activity in the two probiotics groups were increased significantly up to 45 day (P < 0.05), but followed by a decreasing trend by adding Pb. Complement and bactericidal activity were enhanced significantly in the bulgaricus group (P < 0.05). Respiratory burst activity at day 45 in group bulgaricus had significant increase (P < 0.05) and decreased in all groups particularly after Pb exposure (P < 0.05). The achieved data shows that microencapsulation of probiotics with alginate-chitosan may be a suitable method to improve the fish condition against heavy metal.