漂白粉对家蚕中肠产消化酶细菌的影响
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摘要
【目的】本研究以漂白粉液浸渍消毒(以下简称浸消)桑叶饲喂家蚕为处理组,分析了不同组之间家蚕中肠细菌数量的差异;探讨了各组家蚕中肠产消化酶(蛋白酶、淀粉酶、脂肪酶、纤维素酶)细菌的菌株数量、产酶种类及相对产酶能力的差异。【方法】用16S rDNA测序技术对产酶菌株进行了鉴定,比较各组产酶菌种类的不同。【结果】饲喂不同有效氯浓度漂白粉液浸消叶对家蚕中肠细菌的数量、产酶菌数量均有抑制作用,且随有效氯浓度升高抑制作用增强;各处理组产蛋白酶菌株平均相对产酶能力均低于清水组,且清水组肠液中产淀粉酶菌株数明显多于各处理组;取食漂白粉液浸消桑叶后,家蚕中肠产消化酶细菌类群构成有一定变化,清水组和全程0.3%组产酶菌属相对较丰富,有Staphylococcus sp.、Lelliottiasp.和Buttiauxella sp.,其他处理组的产酶菌属相对单一,只有Staphylococcus sp.。【结论】以上结果说明漂白粉对家蚕中肠内细菌及产酶菌的增殖有抑制作用,中肠内产蛋白酶菌的产酶能力和产淀粉酶菌株的数量也受到抑制,对产酶菌类群构成有一定影响。
[Objectives] To investigate the effect of bleach powder on silkworm intestinal bacteria and digestive enzymes,[Methods] Silkworms were either fed mulberry leaves that had been disinfected with bleach powder or untreated mulberry leaves, and their intestinal bacteria were isolated and identified by 16 S rDNA sequencing. [Results] The overall amount of bacteria, and of enzyme-producing bacteria,in the midgut was generally inhibited by bleach powder, regardless of the effective chlorine concentration although there was a positive correlation between the strength of this inhibitory effect and chlorine concentration, Compared to the Control, the abundance of amylase-producing bacteria, as well as the average production of protease, were significantly lower in larvae that had been fed leaves treated with bleach powder, With the exception of the the lowest effective chlorine concentration, 0.3%, exposure to all other concentrations of bleach powder reduced the diversity of mid-gut bacteria to a single genus; Staphylococcus sp., In contrast, the control group and the 0.3% treatment group had relatively diverse enzyme-producing bacteria, including Staphylococcus sp., Lelliottia sp., and Buttiauxella sp. [Conclusion]Both intestinal bacterial community diversity and bacterial digestive enzymes werre significantly inhibited by bleach powder,particularly bacterial protease production and the abundance of amylase-producing bacteria.
[Objectives] To investigate the effect of bleach powder on silkworm intestinal bacteria and digestive enzymes,[Methods] Silkworms were either fed mulberry leaves that had been disinfected with bleach powder or untreated mulberry leaves, and their intestinal bacteria were isolated and identified by 16 S rDNA sequencing. [Results] The overall amount of bacteria, and of enzyme-producing bacteria,in the midgut was generally inhibited by bleach powder, regardless of the effective chlorine concentration although there was a positive correlation between the strength of this inhibitory effect and chlorine concentration, Compared to the Control, the abundance of amylase-producing bacteria, as well as the average production of protease, were significantly lower in larvae that had been fed leaves treated with bleach powder, With the exception of the the lowest effective chlorine concentration, 0.3%, exposure to all other concentrations of bleach powder reduced the diversity of mid-gut bacteria to a single genus; Staphylococcus sp., In contrast, the control group and the 0.3% treatment group had relatively diverse enzyme-producing bacteria, including Staphylococcus sp., Lelliottia sp., and Buttiauxella sp. [Conclusion]Both intestinal bacterial community diversity and bacterial digestive enzymes werre significantly inhibited by bleach powder,particularly bacterial protease production and the abundance of amylase-producing bacteria.
引文
Du YQ,2006.Analysis on the advantages and disadvantages of mulberry leaf disinfection on silkworm production.Proceedings of the Fourth Western Conference on Primary Species.85-86.[杜强远,2006.桑叶消毒对蚕种生产的利弊分析.第四次西部选原种学术研讨会论文集.85-86.]
Gao HJ,Lu GB,Zha CY,2007.Isolation and screening of enzyme producing bacteria from Bombyx mori.Sericulture Science,33(2):228-233.[高绘菊,路国兵,查传勇,等,2007.家蚕肠道产酶菌的分离与筛选.蚕业科学,33(2):228-233.]
Gui ZZ,Chen J,Chen WH,2001.Study on the effect and mechanism of whole silkworm powder(SP)in reducing blood sugar.Sericulture Science,27(2):114-118.[桂仲争,陈杰,陈伟华,2001.全蚕粉(SP)降血糖的作用效果及其机理的研究.蚕业科学,27(2):114-118.]
Li GN,2015.Effects of fluoride stress on intestinal micro-ecological environment of different resistant silkworms.Doctor dissertation.Chongqing:Southwest University.[李冠楠,2015.氟胁迫对不同抗性家蚕肠道微生态环境的影响.博士学位论文.重庆:西南大学.]
Li JH,Fang J,Wu XZ,2000.Plate screening model for alkaline lipase-producing bacteria.Food and Fermentation Industry,26(6):11-14.[李江华,房峻,邬显章,2000.碱性脂肪酶高产菌的平板筛选模型.食品与发酵工业,26(6):11-14.]
Lin RX,Ma YH,Tan TW,2006.Screening and identification of low temperature lipase producing bacteria.Journal of Beijing University of Chemical Technology(Natural Science Edition),33(1):31-35.[林容霞,马延和,谭天伟,2006.低温脂肪酶产生菌的筛选及鉴定.北京化工大学学报(自然科学版),33(1):31-35.]
Reeson AF,Jankovic T,Kasper ML,Rogers S,Austin AD,2003.Application of 16S r DNA-DGGE to examine the microbial ecology associated with a social wasp,Vespula germanica.Insect Molecular Biology,12(1):85.
Rio RVM,Hu Y,Aksoy S,2004.Strategies of the home-team:symbioses exploited for vector-borne disease control.Trends in Microbiology,12(7):425-436.
Schabereitergurtner C,Lubitz W,R?lleke S,2003.Application of broad-range 16S r RNA PCR amplification and DGGEfingerprinting for detection of tick-infecting bacteria.Journal of Microbiological Methods,52(2):251.
Wang JJ,Zhao DY,Liu YG,Ao X,Fan R,Duan ZQ,Liu YP,Chen QX,Jin ZX,Wan YJ,2014.Antagonistic effect of lipopeptide metabolites of Bacillus amyloliquefaciens SWB16strain on Beauveria bassiana.Journal of Microbiology,54(7):778-785.[汪静杰,赵东洋,刘永贵,敖翔,范蕊,段正巧,刘艳萍,陈倩茜,金志雄,万永继,2014.解淀粉芽孢杆菌SWB16菌株脂肽类代谢产物对球孢白僵菌的拮抗作用.微生物学报,54(7):778-785.]
Wei ZJ,Jiang ST,2005.Research progress on silkworm feeding and development.Food Science,26(9):592-596.[魏兆军,姜绍通,2005.蚕饲喂化开发研究进展.食品科学,26(9):592-596.]
Xiang H,Huang YP,2008.The symbiotic relationship between intestinal microbes and insects.Chinese Bulletin of Entomology,45(5):687-693.[相辉,黄勇平,2008.肠道微生物与昆虫的共生关系.昆虫知识,45(5):687-693.]
Yang WJ,Wang ZG,Liu CL,Wei GQ,Zhu BJ,Zou CR,2011.Isolation and identification of amylase producing bacteria from silkworm intestine and cloning and expression of its enzyme gene.Journal of Laser Biology,20(2):219-229.[杨文静,王在贵,刘朝良,魏国清,朱保健,邹昌瑞,2011.家蚕肠道产淀粉酶细菌的分离鉴定及其酶基因的克隆与表达.激光生物学报,20(2):219-229.]
Zou CR,2011.Analysis of intestinal flora of tussah and screening and identification of enzyme-producing bacteria.Doctor dissertation.Anhui Agricultural University.[邹昌瑞,2011.柞蚕肠道菌群分析及产酶菌的筛选与鉴定.博士学位论文.合肥:安徽农业大学.]
Gao HJ,Lu GB,Zha CY,2007.Isolation and screening of enzyme producing bacteria from Bombyx mori.Sericulture Science,33(2):228-233.[高绘菊,路国兵,查传勇,等,2007.家蚕肠道产酶菌的分离与筛选.蚕业科学,33(2):228-233.]
Gui ZZ,Chen J,Chen WH,2001.Study on the effect and mechanism of whole silkworm powder(SP)in reducing blood sugar.Sericulture Science,27(2):114-118.[桂仲争,陈杰,陈伟华,2001.全蚕粉(SP)降血糖的作用效果及其机理的研究.蚕业科学,27(2):114-118.]
Li GN,2015.Effects of fluoride stress on intestinal micro-ecological environment of different resistant silkworms.Doctor dissertation.Chongqing:Southwest University.[李冠楠,2015.氟胁迫对不同抗性家蚕肠道微生态环境的影响.博士学位论文.重庆:西南大学.]
Li JH,Fang J,Wu XZ,2000.Plate screening model for alkaline lipase-producing bacteria.Food and Fermentation Industry,26(6):11-14.[李江华,房峻,邬显章,2000.碱性脂肪酶高产菌的平板筛选模型.食品与发酵工业,26(6):11-14.]
Lin RX,Ma YH,Tan TW,2006.Screening and identification of low temperature lipase producing bacteria.Journal of Beijing University of Chemical Technology(Natural Science Edition),33(1):31-35.[林容霞,马延和,谭天伟,2006.低温脂肪酶产生菌的筛选及鉴定.北京化工大学学报(自然科学版),33(1):31-35.]
Reeson AF,Jankovic T,Kasper ML,Rogers S,Austin AD,2003.Application of 16S r DNA-DGGE to examine the microbial ecology associated with a social wasp,Vespula germanica.Insect Molecular Biology,12(1):85.
Rio RVM,Hu Y,Aksoy S,2004.Strategies of the home-team:symbioses exploited for vector-borne disease control.Trends in Microbiology,12(7):425-436.
Schabereitergurtner C,Lubitz W,R?lleke S,2003.Application of broad-range 16S r RNA PCR amplification and DGGEfingerprinting for detection of tick-infecting bacteria.Journal of Microbiological Methods,52(2):251.
Wang JJ,Zhao DY,Liu YG,Ao X,Fan R,Duan ZQ,Liu YP,Chen QX,Jin ZX,Wan YJ,2014.Antagonistic effect of lipopeptide metabolites of Bacillus amyloliquefaciens SWB16strain on Beauveria bassiana.Journal of Microbiology,54(7):778-785.[汪静杰,赵东洋,刘永贵,敖翔,范蕊,段正巧,刘艳萍,陈倩茜,金志雄,万永继,2014.解淀粉芽孢杆菌SWB16菌株脂肽类代谢产物对球孢白僵菌的拮抗作用.微生物学报,54(7):778-785.]
Wei ZJ,Jiang ST,2005.Research progress on silkworm feeding and development.Food Science,26(9):592-596.[魏兆军,姜绍通,2005.蚕饲喂化开发研究进展.食品科学,26(9):592-596.]
Xiang H,Huang YP,2008.The symbiotic relationship between intestinal microbes and insects.Chinese Bulletin of Entomology,45(5):687-693.[相辉,黄勇平,2008.肠道微生物与昆虫的共生关系.昆虫知识,45(5):687-693.]
Yang WJ,Wang ZG,Liu CL,Wei GQ,Zhu BJ,Zou CR,2011.Isolation and identification of amylase producing bacteria from silkworm intestine and cloning and expression of its enzyme gene.Journal of Laser Biology,20(2):219-229.[杨文静,王在贵,刘朝良,魏国清,朱保健,邹昌瑞,2011.家蚕肠道产淀粉酶细菌的分离鉴定及其酶基因的克隆与表达.激光生物学报,20(2):219-229.]
Zou CR,2011.Analysis of intestinal flora of tussah and screening and identification of enzyme-producing bacteria.Doctor dissertation.Anhui Agricultural University.[邹昌瑞,2011.柞蚕肠道菌群分析及产酶菌的筛选与鉴定.博士学位论文.合肥:安徽农业大学.]