香辣蟹中特征性腐败菌的分离鉴定以及香料的抑制作用研究
|
摘要
目的分析香辣蟹产品的特征性腐败菌,用于指导筛选特定的香辛料配方,以延长产品保质期。方法从自然腐败的市售香辣蟹产品中分离纯化出特征性腐败菌,通过生理生化实验和16SrRNA序列比对法对腐败细菌进行鉴定,选取不同属的腐败菌进行常用香辛料提取物的抑菌效果评价。通过进一步测定香辣蟹在储藏过程中菌落总数和挥发性盐基氮值,评估不同香辛料提取物对香辣蟹的保鲜效果。结果分离所得的15株腐败菌中10株为革兰氏阳性细菌,其余均为革兰氏阴性细菌, 3株属于葡萄球菌属(Staphylococcus. sp), 6株属于白色杆菌属(Leucobacter.sp),3株属于变形杆菌属(Proteus.sp),2株属于普罗威登斯菌(Providencia vermicola),1株属于节杆菌属(Arthrobacter.sp)。不同品种的香辛料对腐败菌生长的抑制作用差异较大,公丁香、八角、香砂、千里香、栀子、草寇对5种指示腐败菌均有体外抑菌作用,研究采用的香辛料配方降低香辣蟹储藏过程中菌落总数和挥发性盐基氮值的增加速度。结论合适的香辛料能延长香辣蟹储藏时间,可以作为香辣蟹天然保鲜剂。
Objective To isolate and purify specific spoilage bacteria in spicy crabs, and screen a formula of spices to prolong the shelf life of spicy crabs. Methods Specific spoilage bacteria were isolated and purified from natural spoiled commercial spicy crabs, and identified by physiological and biochemical characteristics and 16 S rRNA sequence analysis. The bacteriostatic effects of extracts of common spices were evaluated by selecting spoilage bacteria from different genera. By further determining the total colony-forming units and volatile base nitrogen values during the storage of the spicy crab, the preservative effects of the spices were evaluated. Results Totally 15 strains of spoilage bacteria were isolated, including 10 strains of gram-positive bacteria, and the others of gram-negative bacteria, which were 3 Staphylococcus sp. strains, 6 Leucobacter sp. strains, 3 Proteus sp. strains, 2 Providencia vermicola strains, and 1 Arthrobacter sp. strain. The inhibitory effects of different varieties of spices on the growth of spoilage bacteria were different, Eugenia caryophyllata Thunb, Illicium verum Hook, Amomi Semen, Murraya paniculata, Gardenia jasminoides Ellis, and Alpinia hainanensis K. Schum had antibacterial activities against the 5 tested bacteria. The formula used in this study could reduce the total number of colonies and the volatile base nitrogen contents during the storage of spicy crab. Conclusion Suitable spices can prolong the storage time of spicy crab and can be used as a natural preservative for spicy crab.
Objective To isolate and purify specific spoilage bacteria in spicy crabs, and screen a formula of spices to prolong the shelf life of spicy crabs. Methods Specific spoilage bacteria were isolated and purified from natural spoiled commercial spicy crabs, and identified by physiological and biochemical characteristics and 16 S rRNA sequence analysis. The bacteriostatic effects of extracts of common spices were evaluated by selecting spoilage bacteria from different genera. By further determining the total colony-forming units and volatile base nitrogen values during the storage of the spicy crab, the preservative effects of the spices were evaluated. Results Totally 15 strains of spoilage bacteria were isolated, including 10 strains of gram-positive bacteria, and the others of gram-negative bacteria, which were 3 Staphylococcus sp. strains, 6 Leucobacter sp. strains, 3 Proteus sp. strains, 2 Providencia vermicola strains, and 1 Arthrobacter sp. strain. The inhibitory effects of different varieties of spices on the growth of spoilage bacteria were different, Eugenia caryophyllata Thunb, Illicium verum Hook, Amomi Semen, Murraya paniculata, Gardenia jasminoides Ellis, and Alpinia hainanensis K. Schum had antibacterial activities against the 5 tested bacteria. The formula used in this study could reduce the total number of colonies and the volatile base nitrogen contents during the storage of spicy crab. Conclusion Suitable spices can prolong the storage time of spicy crab and can be used as a natural preservative for spicy crab.
引文
[1]周道志,曾凤仙.关于生物保鲜剂在水产品保鲜中的应用分析[J].江西水产科技,2018,(3):47-49.Zhou DZ,Zeng FX.Analysis on the application of biological preservatives in the preservation of aquatic products[J].Jiangxi Fisher Sci Technol,2018,(3):47-49.
[2]刘晨.生物防腐剂及其在食品防腐中的应用[J].食品安全导刊,2017,(15):130.Liu C.Biological preservatives and their application in food preservation[J].China Food Saf Magaz,2017,(15):130.
[3]单珂,郭全友,姜朝军,等.生物保鲜剂在水产品保鲜中的应用[J].食品与发酵科技,2018,54(3):4-8.Shan K,Guo QY,Jiang CJ,et al.Application of biological preservatives in the preservation of aquatic products[J].Food Ferment Technol,2018,54(3):4-8.
[4]刘蒙佳,周强,林海虹.三种天然香辛料液对冷却肉保鲜效果的研究[J].肉类工业,2013,(11):43-48.Liu MJ,Zhou Q,Lin HH.Study on the fresh-keeping effect of three natural spice liquids on cooling meat[J].Meat Ind,2013,(11):43-48.
[5]陈文学,李婷,侯晓东,等.香辛料提取物抑菌作用的研究[J].中国酿造,2007,(9):12-14.Chen WX,Li T,Hou XD,et al.Study on the antibacterial activity of spice extracts[J].China Brew,2007,(9):12-14.
[6]Aydin BD.Investigation of antibacterial effects of some medicinal plants and spices on food pathogens[J].Kafkas Univ Veter Fakult Derg,2008,14(1):83-87.
[7]Beceril R,Gomez-Lus R,Goni P,et al.Combination of analytical and microbiological techniques to study the antimicrobial activity of a new active food packaging containing cinnamon or oregano against E coli and S aureus[J].Anal Bioanalyt Chem,2007,388(5-6):1003-1011.
[8]艾有伟,王丽梅,闫虎山.常用香辛料提取物抑菌活性研究[J].食品工业,2018,39(9):167-170.Ai YW,Wang LM,Yan HS.Study on antibacterial activity of common spice extracts[J].Food Ind,2018,39(9):167-170.
[9]刘小莉,贾洋洋,夏秀东,等.调味料和保鲜剂协同对淡水鱼特征性腐败菌的抑制作用[J].江苏农业科学,2016,44(8):389-391.Liu XL,Jia YY,Xia XD,et al.Inhibition of seasonings and preservatives on the characteristic spoilage bacteria of freshwater fish[J].Jiangsu Agric Sci,2016,44(8):389-391.
[10]GB 4789.2-2016食品安全国家标准食品微生物学检验菌落总数测定[S].GB 4789.2-2016 National food safety standards-Food microbiology testing-Total number of colonies[S].
[11]GB 5009.228-2016食品安全国家标准食品中挥发性盐基氮的测定[S].GB 5009.228-2016 National food safety standard-Determination of volatile base nitrogen in food[S].
[12]于晓慧,葛孟甜,林琳,等.一株常温保藏即食小龙虾优势腐败菌的分离及鉴定[J].肉类工业,2018,(5):33-38.Yu XH,Ge MT,Lin L,et al.Isolation and identification of a dominant spoilage bacteria from ready-to-eat crayfish at room temperature[J].Meat Ind,2018,(5):33-38.
[13]夏秀东,刘小莉,王英,等.常温条件下白鱼中特定腐败菌的鉴定[J].食品科学,2016,37(19):183-189.Xia XD,Liu XL,Wang Y,et al.Identification of specific spoilage bacteria in whitefish under normal temperature conditions[J].Food Sci,2016,37(19):183-189.
[14]高磊,谢晶,叶藻,等.冷鲜鸡腿肉中优势腐败菌的分离鉴定及腐败能力研究[J].食品与发酵工业,2015,41(8):48-53.Gao L,Xie J,Ye Z,et al.Isolation,ss identification and spoilage ability of dominant spoilage bacteria in cold fresh chicken legs[J].Food Ferment Ind,2015,41(8):48-53.
[15]黄佳奇,葛雨珺,向迎春,等.冷藏小黄鱼优势腐败菌致腐能力评价及其对挥发性成分的影响[J].食品工业科技,2018,,39(16):230-235,242.Huang JQ,Ge YJ,Xiang YC,et al.Evaluation of the rotogenic ability of the dominant spoilage bacteria of small yellow croaker and its effect on volatile components[J].Food Ind Sci Technol,2018,39(16):230-235,242.
[16]赵文红,林慧珍,陈海光,等.冰鲜鸭优势腐败菌的鉴定[J].现代食品科技,2012,28(7):728-732.Zhao WH,Lin HZ,Chen HG,et al.Identification of dominant spoilage bacteria in chilled ducks[J].Mod Food Sci Technol,2012,28(7):728-732.
[17]黄现恩.几株微囊藻毒素降解菌和溶藻菌的分离鉴定及作用效果[D].苏州:苏州大学,2015.Huang XE.Isolation,identification and effect of several microcystins-degrading bacteria and algae-lysing bacteria[D].Suzhou:Suzhou University,2015.
[18]高乾坤,杜贺超,赵云飞,等.不同生物保鲜剂对带鱼冷藏保鲜效果的比较[J].食品工业科技,2018,39(22):270-275.Gao QK,Du HC,Zhao YF,et al.Comparison of different biological preservatives on the cold preservation effect of squid[J].Food Ind Sci Technol,2018,39(22):270-275.
[19]刘晓雪,葛菁萍,宋刚,等.Nisin的防腐特性及其在食品工业中的应用[J].黑龙江医药,2016,29(2):195-199.Liu XX,Ge JP,Song G,et al.Antiseptic properties of Nisin and its application in food industry[J].Heilongjiang Med,2016,29(2):195-199.
[20]蔡路昀,马帅,曹爱玲,等.香辛料在水产品保鲜应用中的研究进展[J].食品安全质量检测学报,2015,6(10):3935-3940.Cai LZ,Ma S,Cao AL,et al.Research progress of spices in the application of aquatic products preservation[J].J Food Saf Qual,2015,6(10):3935-3940.
[21]董文霞,王欣.冷藏秋刀鱼的菌相分析及其优势腐败菌的分离鉴定[J].现代食品科技,2016,32(11):139-145,221.Dong WX,Wang X.Analysis of the bacterial phase of refrigerated saury and its isolation and identification of dominant spoilage bacteria[J].Mod Food Sci Technol,2016,32(11):139-145,221.
[2]刘晨.生物防腐剂及其在食品防腐中的应用[J].食品安全导刊,2017,(15):130.Liu C.Biological preservatives and their application in food preservation[J].China Food Saf Magaz,2017,(15):130.
[3]单珂,郭全友,姜朝军,等.生物保鲜剂在水产品保鲜中的应用[J].食品与发酵科技,2018,54(3):4-8.Shan K,Guo QY,Jiang CJ,et al.Application of biological preservatives in the preservation of aquatic products[J].Food Ferment Technol,2018,54(3):4-8.
[4]刘蒙佳,周强,林海虹.三种天然香辛料液对冷却肉保鲜效果的研究[J].肉类工业,2013,(11):43-48.Liu MJ,Zhou Q,Lin HH.Study on the fresh-keeping effect of three natural spice liquids on cooling meat[J].Meat Ind,2013,(11):43-48.
[5]陈文学,李婷,侯晓东,等.香辛料提取物抑菌作用的研究[J].中国酿造,2007,(9):12-14.Chen WX,Li T,Hou XD,et al.Study on the antibacterial activity of spice extracts[J].China Brew,2007,(9):12-14.
[6]Aydin BD.Investigation of antibacterial effects of some medicinal plants and spices on food pathogens[J].Kafkas Univ Veter Fakult Derg,2008,14(1):83-87.
[7]Beceril R,Gomez-Lus R,Goni P,et al.Combination of analytical and microbiological techniques to study the antimicrobial activity of a new active food packaging containing cinnamon or oregano against E coli and S aureus[J].Anal Bioanalyt Chem,2007,388(5-6):1003-1011.
[8]艾有伟,王丽梅,闫虎山.常用香辛料提取物抑菌活性研究[J].食品工业,2018,39(9):167-170.Ai YW,Wang LM,Yan HS.Study on antibacterial activity of common spice extracts[J].Food Ind,2018,39(9):167-170.
[9]刘小莉,贾洋洋,夏秀东,等.调味料和保鲜剂协同对淡水鱼特征性腐败菌的抑制作用[J].江苏农业科学,2016,44(8):389-391.Liu XL,Jia YY,Xia XD,et al.Inhibition of seasonings and preservatives on the characteristic spoilage bacteria of freshwater fish[J].Jiangsu Agric Sci,2016,44(8):389-391.
[10]GB 4789.2-2016食品安全国家标准食品微生物学检验菌落总数测定[S].GB 4789.2-2016 National food safety standards-Food microbiology testing-Total number of colonies[S].
[11]GB 5009.228-2016食品安全国家标准食品中挥发性盐基氮的测定[S].GB 5009.228-2016 National food safety standard-Determination of volatile base nitrogen in food[S].
[12]于晓慧,葛孟甜,林琳,等.一株常温保藏即食小龙虾优势腐败菌的分离及鉴定[J].肉类工业,2018,(5):33-38.Yu XH,Ge MT,Lin L,et al.Isolation and identification of a dominant spoilage bacteria from ready-to-eat crayfish at room temperature[J].Meat Ind,2018,(5):33-38.
[13]夏秀东,刘小莉,王英,等.常温条件下白鱼中特定腐败菌的鉴定[J].食品科学,2016,37(19):183-189.Xia XD,Liu XL,Wang Y,et al.Identification of specific spoilage bacteria in whitefish under normal temperature conditions[J].Food Sci,2016,37(19):183-189.
[14]高磊,谢晶,叶藻,等.冷鲜鸡腿肉中优势腐败菌的分离鉴定及腐败能力研究[J].食品与发酵工业,2015,41(8):48-53.Gao L,Xie J,Ye Z,et al.Isolation,ss identification and spoilage ability of dominant spoilage bacteria in cold fresh chicken legs[J].Food Ferment Ind,2015,41(8):48-53.
[15]黄佳奇,葛雨珺,向迎春,等.冷藏小黄鱼优势腐败菌致腐能力评价及其对挥发性成分的影响[J].食品工业科技,2018,,39(16):230-235,242.Huang JQ,Ge YJ,Xiang YC,et al.Evaluation of the rotogenic ability of the dominant spoilage bacteria of small yellow croaker and its effect on volatile components[J].Food Ind Sci Technol,2018,39(16):230-235,242.
[16]赵文红,林慧珍,陈海光,等.冰鲜鸭优势腐败菌的鉴定[J].现代食品科技,2012,28(7):728-732.Zhao WH,Lin HZ,Chen HG,et al.Identification of dominant spoilage bacteria in chilled ducks[J].Mod Food Sci Technol,2012,28(7):728-732.
[17]黄现恩.几株微囊藻毒素降解菌和溶藻菌的分离鉴定及作用效果[D].苏州:苏州大学,2015.Huang XE.Isolation,identification and effect of several microcystins-degrading bacteria and algae-lysing bacteria[D].Suzhou:Suzhou University,2015.
[18]高乾坤,杜贺超,赵云飞,等.不同生物保鲜剂对带鱼冷藏保鲜效果的比较[J].食品工业科技,2018,39(22):270-275.Gao QK,Du HC,Zhao YF,et al.Comparison of different biological preservatives on the cold preservation effect of squid[J].Food Ind Sci Technol,2018,39(22):270-275.
[19]刘晓雪,葛菁萍,宋刚,等.Nisin的防腐特性及其在食品工业中的应用[J].黑龙江医药,2016,29(2):195-199.Liu XX,Ge JP,Song G,et al.Antiseptic properties of Nisin and its application in food industry[J].Heilongjiang Med,2016,29(2):195-199.
[20]蔡路昀,马帅,曹爱玲,等.香辛料在水产品保鲜应用中的研究进展[J].食品安全质量检测学报,2015,6(10):3935-3940.Cai LZ,Ma S,Cao AL,et al.Research progress of spices in the application of aquatic products preservation[J].J Food Saf Qual,2015,6(10):3935-3940.
[21]董文霞,王欣.冷藏秋刀鱼的菌相分析及其优势腐败菌的分离鉴定[J].现代食品科技,2016,32(11):139-145,221.Dong WX,Wang X.Analysis of the bacterial phase of refrigerated saury and its isolation and identification of dominant spoilage bacteria[J].Mod Food Sci Technol,2016,32(11):139-145,221.