不同干露条件单环刺螠的耐受性及对体表黏液特性的影响
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摘要
为了研究单环刺螠Urechis unicinctus在不同温度(15、20、25、30℃)的湿沙无水和湿沙有水干露条件下的耐受性、体表黏液分泌量及成分含量的变化,试验采用离心管提取平均体质量为2.75 g的单环刺螠体表黏液,采用苯酚—硫酸法和总蛋白测定试剂盒分别检测黏液中多糖和蛋白质含量,同时研究其黏液对金黄色葡萄球菌Staphylococcus aureus、灿烂弧菌Vibrio splendidus、奇艺变形杆菌Bacillus mirabilis、产气肠杆菌Enterobacter aerogenes、阴沟肠杆菌Aerobacter cloacae、哈维氏弧菌Vibrio harveyi、副溶血弧菌Vibrio parahaemolyticus、鳗弧菌Vibrio anguillarum、藤黄微球菌Micrococcus luteus等常见水产动物病原菌的抗菌性,采用抑菌圈法通过比较抑菌圈直径以判断抗菌效果。结果表明:单环刺螠体表黏液分泌量随温度升高而降低,湿沙无水干露条件下的单环刺螠耐受性要强于湿沙有水;15℃条件下,10只单环刺螠湿沙无水和湿沙有水型分别在1 min(3.33 mL±0.05 mL)和10 min(2.85 mL±0.04 mL)时黏液分泌量较高;湿沙无水条件下多糖(1.04μg/mL±0.10μg/mL)及蛋白质(65.81 mg/mL±2.22 mg/mL)含量均在30 min时最高,且随干露时间的延长均呈先升高后降低的趋势;体表黏液对所测9种病原菌的抑菌圈直径小于10 mm,均无抗菌性。研究表明,2.75 g规格的单环刺螠,在15℃湿沙无水干露条件下耐受性可达48 h以上。
The polysaccharide and protein contents and secretion were detected in surface mucus of echiuran worm Urechis unicinctus with body weight of 2.75 g in a wet sand plastic box with and without water at temperature of 15, 20, 25, and 30 ℃ by phenol-sulfuric acid method and total protein assay kit, and the antibacterial activity of mucus to routine pathogens Staphylococcus aureus, Vibrio splendidus, Bacillus mirabilis, Enterobacter aerogenes, Aerobacter cloacae, V.harveyi, V.parahaemolyticus, V.anguillarum, and Micrococcus luteus in aquatic animals was tested by comparing the diameter of the bacteriostatic zone to evaluate air exposure tolerance and characteristics of surface mucus in echiuran worm in wet sand with and without water. It was found that the surface mucus secretion was decreased with the increase in temperature, and that there was stronger tolerance to air exposure in the echiuran worm in wet sand without water than that in the echiuran worm in wet sand with water, the maximal surface mucus secretion in 10 individuals in 10 min(2.85 mL±0.04 mL) in echiuran worm in wet sand with water and in 1 min(3.33 mL±0.05 mL) in echiuran worm in wet sand without water at 15 ℃. The maximal contents of polysaccharide(1.04 μg/mL±0.10 μg/mL) and protein(65.81 mg/mL±2.22 mg/mL) were observed in 30 min in echiuran worm in wet sand without water, with a tendency of first increase and then decrease. No antimicrobial activity of the mucus to the tested 9 pathogens was found. The findings indicate that the echiuran worm with body weight of 2.75 g can tolerate air exposure for more than 48 hours under the condition of wet sand without water at 15 ℃.
The polysaccharide and protein contents and secretion were detected in surface mucus of echiuran worm Urechis unicinctus with body weight of 2.75 g in a wet sand plastic box with and without water at temperature of 15, 20, 25, and 30 ℃ by phenol-sulfuric acid method and total protein assay kit, and the antibacterial activity of mucus to routine pathogens Staphylococcus aureus, Vibrio splendidus, Bacillus mirabilis, Enterobacter aerogenes, Aerobacter cloacae, V.harveyi, V.parahaemolyticus, V.anguillarum, and Micrococcus luteus in aquatic animals was tested by comparing the diameter of the bacteriostatic zone to evaluate air exposure tolerance and characteristics of surface mucus in echiuran worm in wet sand with and without water. It was found that the surface mucus secretion was decreased with the increase in temperature, and that there was stronger tolerance to air exposure in the echiuran worm in wet sand without water than that in the echiuran worm in wet sand with water, the maximal surface mucus secretion in 10 individuals in 10 min(2.85 mL±0.04 mL) in echiuran worm in wet sand with water and in 1 min(3.33 mL±0.05 mL) in echiuran worm in wet sand without water at 15 ℃. The maximal contents of polysaccharide(1.04 μg/mL±0.10 μg/mL) and protein(65.81 mg/mL±2.22 mg/mL) were observed in 30 min in echiuran worm in wet sand without water, with a tendency of first increase and then decrease. No antimicrobial activity of the mucus to the tested 9 pathogens was found. The findings indicate that the echiuran worm with body weight of 2.75 g can tolerate air exposure for more than 48 hours under the condition of wet sand without water at 15 ℃.
引文
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[2] 刘峰,孙涛,纪元,等.单环刺螠生物学及生态学研究进展[J].海洋科学,2017,41(10):125-131.
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[8] 张婷,史晋绒,宋柯,等.鱼类体表黏液分泌机制·组分与功能研究进展[J].安徽农业科学,2014,42(22):7445-7448,7458.
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[11] Zamzow J,Losey G S.Ultraviolet radiation absorbance by coral reef fish mucus:photo-protection and visual communication[J].Environmental Biology of Fishes,2002,63(1):41-47.
[12] 潘高,刘念,许云,等.泥鳅黏液防生物污损作用研究[J].淡水渔业,2014,44(1):98-101.
[13] 许星鸿,霍伟,孟宵,等.单环刺螠人工育苗及养殖技术[J].科学养鱼,2016(2):53-55.
[14] Hidu H,Chapman S,Mook W.Overwintering American oyster seed by cold humid air storage[J].Journal of Shellfish Research,1988,7(1):47-50.
[15] Seaman M N L.Survival and aspects of metabolism in oysters,Crassostrea gigas,during and after prolonged air storage[J].Aquaculture,1991,93(4):389-395.
[16] Hemmerich S.Glycomics:coming of age across the globe[J].Drug Discovery Today,2005,10(5):307-309.
[17] 孙虎山.长竹蛏苗的潜沙及耐干露能力研究[J].烟台师范学院学报:自然科学版,1992,8(1-2):67-69,73.
[18] 林景超,刘旭佳,沈夏霜,等.方格星虫中培苗对温度、盐度和干露的耐受性试验[J].水产科学,2016,35(3):267-271.
[19] 刘旭绪,张秀梅,覃乐政,等.干露时长及温度对魁蚶幼贝潜沙行为及呼吸代谢的影响[J].中国海洋大学学报:自然科学版,2017,47(3):19-26.
[20] 于瑞海,辛荣,赵强,等.海湾扇贝不同发育阶段耐干露的研究[J].海洋科学,2007,31(6):6-9.
[21] 孙煜阳,童巧琼,王文,等.厚壳贻贝在不同温度下的干露耐受性研究[J].生物学杂志,2017,34(4):42-46.
[22] 李忠泓,王国栋.硬壳蛤稚贝对淡水浸泡、干露和低温的耐受能力[J].水产科学,2004,23(6):14-16.
[23] 段亚飞,董宏标,王芸,等.干露胁迫对日本囊对虾抗氧化酶活性的影响[J].南方水产科学,2015,11(4):102-108.
[24] 田相利,何瑞鹏,钱圆,等.干露胁迫对刺参体壁非特异性免疫的影响[J].河北渔业,2014(7):21-26,35.
[25] 平洪领,李玉全.干露对天津厚蟹水分代谢和死亡率的影响[J].湖北农业科学,2014,53(18):4377-4380.
[26] 白艳龙,谭昭仪,邸向乾,等.黄颡鱼无水保活技术研究[J].食品工业科技,2013,34(1):334-337.
[27] 肖友翔,霍忠明,张学开,等.干露对日本海神蛤稚贝生长和存活的影响[J].大连海洋大学学报,2016,31(5):482-487.
[28] Zhang Yongjun,Zhang Xiaoshuan,Nga M T T,et al.Development and evaluation of key ambient factors online monitoring system in live Urechis unicinctus transportation strategies[J].Computers and Electronics in Agriculture,2018,145:43-52.
[29] Winn H E.Formation of a mucous envelope at night by parrot fishes[J].Zoologica,1955(40):145-148.
[30] Wiley J W.Observations on the use of mucus envelopes by the California sheephead,Pimelometopon pulchrum,on Southern California rock reefs[J].Copeia,1974,1974(3):789-790.
[31] 尹绍武,周工健,刘筠.黄鳝血清和体表粘液蛋白的比较研究[J].激光生物学报,2003,12(2):128-131.
[32] 邱进,潘连德,张帅.松江鲈鱼体表黏液抑菌效果研究[J].生物技术通报,2014(9):195-200.
[33] 李联泰,安贤惠,胡江,等.黄鳝皮肤黏液抗菌肽的分离纯化及其部分特性研究[J].渔业科学进展,2011,32(2):27-33.
[34] 郑风荣,徐宗军,张永强,等.星斑川鲽免疫相关组织抗菌活性的研究[J].中国渔业质量与标准,2015,5(3):63-69.
[2] 刘峰,孙涛,纪元,等.单环刺螠生物学及生态学研究进展[J].海洋科学,2017,41(10):125-131.
[3] 杨桂文,安利国,孙忠军.单环刺螠营养成分分析[J].海洋科学,1999(6):13-14.
[4] 李诺,宋淑莲,唐永政,等.单环刺螠增养殖生物学的研究[J].齐鲁渔业,1998,15(1):11-14.
[5] 王力勇,胡丽萍,赵强,等.单环刺螠亲体运输及暂养技术研究[J].中国水产,2017(4):93-94.
[6] 杨玉品.单环刺螠(Urechis unicinctus)多糖的分离纯化和结构研究[D].青岛:中国海洋大学,2011:1-76.
[7] 李诺,宋淑莲,唐永政,等.单环刺螠体壁氨基酸组分与含量的分析[J].齐鲁渔业,2000,17(5):26-27.
[8] 张婷,史晋绒,宋柯,等.鱼类体表黏液分泌机制·组分与功能研究进展[J].安徽农业科学,2014,42(22):7445-7448,7458.
[9] Shephard K L.Functions for fish mucus[J].Reviews in Fish Biology and Fisheries,1994,4(4):401-429.
[10] Yuan Chunying,Liu Ping,Han Xu,et al.Hypoglycemic effects of glycosaminoglycan from Urechis unicinctus in diabetic mice[J].Journal of Medicinal Food,2015,18(2):190-194.
[11] Zamzow J,Losey G S.Ultraviolet radiation absorbance by coral reef fish mucus:photo-protection and visual communication[J].Environmental Biology of Fishes,2002,63(1):41-47.
[12] 潘高,刘念,许云,等.泥鳅黏液防生物污损作用研究[J].淡水渔业,2014,44(1):98-101.
[13] 许星鸿,霍伟,孟宵,等.单环刺螠人工育苗及养殖技术[J].科学养鱼,2016(2):53-55.
[14] Hidu H,Chapman S,Mook W.Overwintering American oyster seed by cold humid air storage[J].Journal of Shellfish Research,1988,7(1):47-50.
[15] Seaman M N L.Survival and aspects of metabolism in oysters,Crassostrea gigas,during and after prolonged air storage[J].Aquaculture,1991,93(4):389-395.
[16] Hemmerich S.Glycomics:coming of age across the globe[J].Drug Discovery Today,2005,10(5):307-309.
[17] 孙虎山.长竹蛏苗的潜沙及耐干露能力研究[J].烟台师范学院学报:自然科学版,1992,8(1-2):67-69,73.
[18] 林景超,刘旭佳,沈夏霜,等.方格星虫中培苗对温度、盐度和干露的耐受性试验[J].水产科学,2016,35(3):267-271.
[19] 刘旭绪,张秀梅,覃乐政,等.干露时长及温度对魁蚶幼贝潜沙行为及呼吸代谢的影响[J].中国海洋大学学报:自然科学版,2017,47(3):19-26.
[20] 于瑞海,辛荣,赵强,等.海湾扇贝不同发育阶段耐干露的研究[J].海洋科学,2007,31(6):6-9.
[21] 孙煜阳,童巧琼,王文,等.厚壳贻贝在不同温度下的干露耐受性研究[J].生物学杂志,2017,34(4):42-46.
[22] 李忠泓,王国栋.硬壳蛤稚贝对淡水浸泡、干露和低温的耐受能力[J].水产科学,2004,23(6):14-16.
[23] 段亚飞,董宏标,王芸,等.干露胁迫对日本囊对虾抗氧化酶活性的影响[J].南方水产科学,2015,11(4):102-108.
[24] 田相利,何瑞鹏,钱圆,等.干露胁迫对刺参体壁非特异性免疫的影响[J].河北渔业,2014(7):21-26,35.
[25] 平洪领,李玉全.干露对天津厚蟹水分代谢和死亡率的影响[J].湖北农业科学,2014,53(18):4377-4380.
[26] 白艳龙,谭昭仪,邸向乾,等.黄颡鱼无水保活技术研究[J].食品工业科技,2013,34(1):334-337.
[27] 肖友翔,霍忠明,张学开,等.干露对日本海神蛤稚贝生长和存活的影响[J].大连海洋大学学报,2016,31(5):482-487.
[28] Zhang Yongjun,Zhang Xiaoshuan,Nga M T T,et al.Development and evaluation of key ambient factors online monitoring system in live Urechis unicinctus transportation strategies[J].Computers and Electronics in Agriculture,2018,145:43-52.
[29] Winn H E.Formation of a mucous envelope at night by parrot fishes[J].Zoologica,1955(40):145-148.
[30] Wiley J W.Observations on the use of mucus envelopes by the California sheephead,Pimelometopon pulchrum,on Southern California rock reefs[J].Copeia,1974,1974(3):789-790.
[31] 尹绍武,周工健,刘筠.黄鳝血清和体表粘液蛋白的比较研究[J].激光生物学报,2003,12(2):128-131.
[32] 邱进,潘连德,张帅.松江鲈鱼体表黏液抑菌效果研究[J].生物技术通报,2014(9):195-200.
[33] 李联泰,安贤惠,胡江,等.黄鳝皮肤黏液抗菌肽的分离纯化及其部分特性研究[J].渔业科学进展,2011,32(2):27-33.
[34] 郑风荣,徐宗军,张永强,等.星斑川鲽免疫相关组织抗菌活性的研究[J].中国渔业质量与标准,2015,5(3):63-69.