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用于鱼类口服免疫的藻酸盐微囊化:释放特性、体外肠道摄取及在大西洋鲑(Salmo salar L.)体内的给药

Alginate Microencapsulation for Oral Immunisation of Finfish: Release Characteristics, Ex Vivo Intestinal Uptake and In Vivo Administration in Atlantic Salmon, Salmo salar L.

作者信息

Ghosh Bikramjit, Nowak Barbara F, Bridle Andrew R

机构信息

Institute for Marine and Antarctic Studies, University of Tasmania, Locked Bag 1370, Launceston, Tasmania, 7250, Australia.

出版信息

Mar Biotechnol (NY). 2015 Dec;17(6):841-53. doi: 10.1007/s10126-015-9663-7. Epub 2015 Sep 26.

Abstract

This study examined the feasibility of alginate microcapsules manufactured using a low-impact technology and reagents to protect orally delivered immunogens for use as immunoprophylactics for fish. Physical characteristics and protein release kinetics of the microcapsules were examined at different pH and temperature levels using a microencapsulated model protein, bovine serum albumin (BSA). Impact of the microencapsulation process on contents was determined by analysing change in bioactivity of microencapsulated lysozyme. Feasibility of the method for oral immunoprophylaxis of finfish was assessed using FITC-labelled microcapsules. These were applied to distal intestinal explants of Atlantic salmon (Salmo salar) to investigate uptake ex vivo. Systemic distribution of microcapsules was investigated by oral administration of FITC-labelled microcapsules to Atlantic salmon fry by incorporating into feed. The microcapsules produced were structurally robust and retained surface integrity, with a modal size distribution of 250-750 nm and a tendency to aggregate. Entrapment efficiency of microencapsulation was 51.2 % for BSA and 43.2 % in the case of lysozyme. Microcapsules demonstrated controlled release of protein, which increased with increasing pH or temperature, and the process had no significant negative effect on bioactivity of lysozyme. Uptake of fluorescent-labelled microcapsules was clearly demonstrated by intestinal explants over a 24-h period. Evidence of microcapsules was found in the intestine, spleen, kidney and liver of fry following oral administration. Amenability of the microcapsules to intestinal uptake and distribution reinforced the strong potential for use of this microencapsulation method in oral immunoprophylaxis of finfish using sensitive immunogenic substances.

摘要

本研究考察了使用低影响技术和试剂制造藻酸盐微胶囊以保护口服免疫原用作鱼类免疫预防剂的可行性。使用微囊化模型蛋白牛血清白蛋白(BSA),在不同pH和温度水平下检测微胶囊的物理特性和蛋白质释放动力学。通过分析微囊化溶菌酶生物活性的变化来确定微囊化过程对其内容物的影响。使用异硫氰酸荧光素(FITC)标记的微胶囊评估该方法用于硬骨鱼口服免疫预防的可行性。将这些微胶囊应用于大西洋鲑(Salmo salar)的远端肠外植体以研究离体摄取情况。通过将FITC标记的微胶囊掺入饲料中口服给予大西洋鲑鱼苗,研究微胶囊的全身分布。所制备的微胶囊结构坚固且保持表面完整性,模态尺寸分布为250 - 750 nm且有聚集趋势。BSA的微囊化包封效率为51.2%,溶菌酶的包封效率为43.2%。微胶囊表现出蛋白质的控释,其随pH或温度升高而增加,且该过程对溶菌酶的生物活性无显著负面影响。肠外植体在24小时内清楚地证明了对荧光标记微胶囊的摄取。口服给药后,在鱼苗的肠道、脾脏、肾脏和肝脏中发现了微胶囊的踪迹。微胶囊易于被肠道摄取和分布,这增强了使用这种微囊化方法利用敏感免疫原性物质对硬骨鱼进行口服免疫预防的巨大潜力。

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