天梯山石窟壁画保存环境中空气细菌的季节性变化
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摘要
【背景】微生物侵蚀是古代壁画常见生物病害,影响壁画的长久保存和安全陈展。空气微生物作为壁画病害菌的主要来源,近年在文物赋存环境监测和预防性保护中引起广泛重视。【目的】对天梯山石窟壁画的2处保存地,即天梯山原址和武威西夏博物馆壁画保存环境中的空气细菌浓度、群落结构及其季节变化规律进行分析。【方法】利用生物气溶胶采样器,在2016年春、夏、秋、冬4季分别采集各位点空气样品;基于传统培养方法获得空气中细菌浓度及纯培养菌株;通过提取细菌基因组DNA、扩增其16S rRNA基因、测序和系统发生关系分析等技术研究不同位点细菌群落时空动态变化规律;结合环境监测数据,分析影响文化遗产地空气细菌群落变化的主要因素。【结果】空气可培养细菌的总浓度在16.7-1 451.8 CFU/m3范围内变动。原址第18窟和第13窟,各季节细菌浓度无显著性差异,且呈明显季节性变动规律,总体特征为夏秋季低,冬春季高。西夏博物馆外空气细菌浓度在各季节均高于库房内,冬季最高。本研究共鉴定出19个细菌属,隶属于4个门;其中不动杆菌属(Acinetobacter)、节杆菌属(Arthrobacter)、芽孢杆菌属(Bacillus)、考克氏菌属(Kocuria)、短波单胞菌属(Brevundimonas)、肉食杆菌属(Carnobacterium)、 Pseudoclavibacter和薄层菌属(Hymenobacter)为优势属。【结论】天梯山石窟空气细菌群落结构具有明显的时空分布特征;相对湿度、温度及季节性降水均会影响其变化;鉴定得到部分种属具备引起壁画生物腐蚀的潜势;本研究可为当地开展遗址和馆藏环境中文物预防性保护提供本底资料。
[Background] Microbial deterioration is a common biological disease in ancient wall paintings,which seriously threaten the long-term safety preservation and exhibition of the wall paintings. As the main source of the harmful microorganisms to wall paintings, airborne microorganisms have attracted extensive attention in the monitoring and preventive conservation of cultural relics in recent years.[Objective] Systematic survey of culturable airborne bacteria, including the concentration, community structure and seasonal variation, was carried out in environments of wall paintings preserved at Tiantishan Grottoes and Western Xia Museum, China. [Methods] Bio-aerosol sampler containing R2 A agar was used for sampling in all four seasons in 2016. Traditional culture-based method to acquire the airborne bacterial concentration information and purified strains; by the extraction of genomic DNA, amplification of bacteria 16 S rRNA gene region, sequencing, and phylogenetic analysis, thereafter the bacteria community composition and distribution characteristics of different study sites were clarified. Combined with environmental monitoring data, to disclosure the main factors which responsible for the dynamic changes of the airborne bacteria at the site. [Results] The concentration of culturable airborne bacteria was in a range from 16.7 to 1 451.8 CFU/m3. There was no significant difference in bacteria concentration between the Cave 18 and Cave 13, namely, the two sites at the Tiantishan Grottoes, with the obvious characteristics of the seasonal variation, briefly speaking, Winter and Spring were higher than Summer and Autumn.Meanwhile, there was significant difference in bacteria concentration between the inside and outside of the Xixia Museum, the square outside of the museum were far more than the inside of the museum at the four seasons, particularly in the winter. A total of 19 bacteria genera that affiliated to four phylum were detected, among which, Acinetobacter, Arthrobacter, Bacillus, Kocuria, Brevundimonas, Carnobacterium,Pseudoclavibacter and Hymenobacter were the dominant ones. [Conclusion] The airborne bacterial community structures of the Tiantishan Grottoes showed a distinct characteristic of the seasonal variation and spatial distribution; RH, temperature and seasonal rainfall were all have influences on the airborne bacteria distribution; some of the genera have potential of causing biodeterioration of the ancient wall paintings at this site; this study could provide supporting information for the preventive protection of cultural relics that preserved at local site and museums.
[Background] Microbial deterioration is a common biological disease in ancient wall paintings,which seriously threaten the long-term safety preservation and exhibition of the wall paintings. As the main source of the harmful microorganisms to wall paintings, airborne microorganisms have attracted extensive attention in the monitoring and preventive conservation of cultural relics in recent years.[Objective] Systematic survey of culturable airborne bacteria, including the concentration, community structure and seasonal variation, was carried out in environments of wall paintings preserved at Tiantishan Grottoes and Western Xia Museum, China. [Methods] Bio-aerosol sampler containing R2 A agar was used for sampling in all four seasons in 2016. Traditional culture-based method to acquire the airborne bacterial concentration information and purified strains; by the extraction of genomic DNA, amplification of bacteria 16 S rRNA gene region, sequencing, and phylogenetic analysis, thereafter the bacteria community composition and distribution characteristics of different study sites were clarified. Combined with environmental monitoring data, to disclosure the main factors which responsible for the dynamic changes of the airborne bacteria at the site. [Results] The concentration of culturable airborne bacteria was in a range from 16.7 to 1 451.8 CFU/m3. There was no significant difference in bacteria concentration between the Cave 18 and Cave 13, namely, the two sites at the Tiantishan Grottoes, with the obvious characteristics of the seasonal variation, briefly speaking, Winter and Spring were higher than Summer and Autumn.Meanwhile, there was significant difference in bacteria concentration between the inside and outside of the Xixia Museum, the square outside of the museum were far more than the inside of the museum at the four seasons, particularly in the winter. A total of 19 bacteria genera that affiliated to four phylum were detected, among which, Acinetobacter, Arthrobacter, Bacillus, Kocuria, Brevundimonas, Carnobacterium,Pseudoclavibacter and Hymenobacter were the dominant ones. [Conclusion] The airborne bacterial community structures of the Tiantishan Grottoes showed a distinct characteristic of the seasonal variation and spatial distribution; RH, temperature and seasonal rainfall were all have influences on the airborne bacteria distribution; some of the genera have potential of causing biodeterioration of the ancient wall paintings at this site; this study could provide supporting information for the preventive protection of cultural relics that preserved at local site and museums.
引文
[1]Gonzalez JM,Portillo MC,Saiz-Jimenez C.Diverse microbial communities and the conservation of prehistoric paintings[J].Microbe,2008,3:72-77
[2]Schabereiter-Gurtner C,Saiz-Jimenez C,Pi?ar G,et al.Phylogenetic diversity of bacteria associated with Paleolithic paintings and surrounding rock walls in two Spanish caves(Llonín and La Garma)[J].FEMSMicrobiology Ecology,2004,47(2):235-247
[3]Ma YT,Zhang H,Du Y,et al.The community distribution of bacteria and fungi on ancient wall paintings of the Mogao Grottoes[J].Scientific Reports,2015,5:7752
[4]Duan YL,Wu FS,Wang WF,et al.The microbial community characteristics of ancient painted sculptures in Maijishan Grottoes,China[J].PLoS One,2017,12(7):e0179718
[5]Bastian F,Jurado V,NovákováA,et al.The microbiology of Lascaux Cave[J].Microbiology,2010,156(3):644-652
[6]Dupont J,Jacquet C,Dennetière B,et al.Invasion of the French Paleolithic painted cave of Lascaux by members of the Fusarium solani species complex[J].Mycologia,2007,99(4):526-533
[7]Portillo MC,Saiz-Jimenez C,Gonzalez JM.Molecular characterization of total and metabolically active bacterial communities of‘‘white colonizations’’in the Altamira Cave,Spain[J].Research in Microbiology,2009,160(1):41-47
[8]Schabereiter-Gurtner C,Saiz-Jimenez C,Pi?ar G,et al.Phylogenetic 16S rRNA analysis reveals the presence of complex and partly unknown bacterial communities in Tito Bustillo cave,Spain,and on its Palaeolithic paintings[J].Environmental Microbiology,2002,4(7):392-400
[9]Song LH,Song WM,Shi W,et al.Health effects of atmospheric microbiological pollution on respiratory system among children in Shanghai[J].Journal of Environment and Health,2000,17(3):135-138(in Chinese)宋凌浩,宋伟民,施玮,等.上海市大气微生物污染对儿童呼吸系统健康影响的研究[J].环境与健康杂志,2000,17(3):135-138
[10]Wang WF,Zhao LY,Pei QQ,et al.Exploration and practice of conservation of wall paintings in the museum:the case of conservation of wall paintings collected in the Gansu Museum from Tiantishan Grottoes,Wuwei[J].Sciences of Conservation and Archaeology,2015,27(4):101-112(in Chinese)汪万福,赵林毅,裴强强,等.馆藏壁画保护理论探索与实践--以甘肃省博物馆藏武威天梯山石窟壁画的保护修复为例[J].文物保护与考古科学,2015,27(4):101-112
[11]Bonazza A,de Nuntiis P,Mandrioli P,et al.Aerosol impact on cultural heritage:deterioration processes and strategies for preventive conservation[A]//Tomasi C,Fuzzi S,Kokhanovsky A.Atmospheric Aerosols:Life Cycles and Effects on Air Quality and Climate[M].Weinheim:Wiley,2016:645-670
[12]Wang WF,Ma YT,Ma X,et al.Seasonal variations of airborne bacteria in the Mogao Grottoes,Dunhuang,China[J].International Biodeterioration&Biodegradation,2010,64(4):309-315
[13]Wang WF,Ma X,Ma YT,et al.Seasonal dynamics of airborne fungi in different caves of the Mogao Grottoes,Dunhuang,China[J].International Biodeterioration&Biodegradation,2010,64(6):461-466
[14]Docampo S,Trigo MM,Recio M,et al.Fungal spore content of the atmosphere of the cave of Nerja(southern Spain):diversity and origin[J].Science of the Total Environment,2011,409(4):835-843
[15]Wu WT,He HP,Yan L,et al.The isolation and identification of bacteria in the air of Capital Museum and their significance for protection of cultural relics[J].Sciences of Conservation and Archaeology,2012,24(1):76-82(in Chinese)武望婷,何海平,闫丽,等.首都博物馆空气中细菌的分离鉴定及在文物保护中的意义[J].文物保护与考古科学,2012,24(1):76-82
[16]E J,Wu FS,Wang WF,et al.Monitoring and research on microbes in the environment of the wall paintings in No.5of the Wei and Jin Tombs[J].Dunhuang Research,2013(6):109-116(in Chinese)俄军,武发思,汪万福,等.魏晋五号壁画墓保存环境中空气微生物监测研究[J].敦煌研究,2013(6):109-116
[17]Wang WF,Ma YT,Ma X,et al.Diversity and seasonal dynamics of airborne bacteria in the Mogao Grottoes,Dunhuang,China[J].Aerobiologia,2012,28(1):27-38
[18]Tang H,Fan WQ,Wang C,et al.A dynamic study on the species and quantity of microorganisms in the micro-environment,Chongqing China Three Gorges Museum[J].Sciences of Conservation and Archaeology,2017,29(1):35-43(in Chinese)唐欢,范文奇,王春,等.重庆中国三峡博物馆小环境空气微生物种属与数量的动态研究[J].文物保护与考古科学,2017,29(1):35-43
[19]Gaüzère C,Moletta-Denat M,Blanquart H,et al.Stability of airborne microbes in the Louvre Museum over time[J].Indoor Air,2014,24(1):29-40
[20]Gaüzère C,Moletta-Denat M,Bousta F,et al.Airborne microbiology in museum through molecular approach:preliminary study[C].Indoor Air Quality,2010
[21]Ruga L,Orlandi F,Romano B,et al.The assessment of fungal bioaerosols in the crypt of St.Peter in Perugia(Italy)[J].International Biodeterioration&Biodegradation,2015,98:121-130
[22]Gaüzère C,Moletta-Denat M,Bousta F,et al.Reliable procedure for molecular analysis of airborne microflora in three indoor environments:an office and two different museum contexts[J].Clean-Soil,Air,Water,2013,41(3):226-234
[23]Anaya M,Borrego SF,Gámez E,et al.Viable fungi in the air of indoor environments of the National Archive of the Republic of Cuba[J].Aerobiologia,2016,32(3):513-527
[24]Lazaridis M,Katsivela E,Kopanakis I,et al.Indoor/outdoor particulate matter concentrations and microbial load in cultural heritage collections[J].Heritage Science,2015,3:34
[25]Tanaka D,Terada Y,Nakashima T,et al.Seasonal variations in airborne bacterial community structures at a suburban site of central Japan over a 1-year time period using PCR-DGGEmethod[J].Aerobiologia,2015,31(2):143-157
[26]Xue LG,Jiang JR,Famous E.Progress in research and monitoring of urban airborne microbes[J].Environmental Engineering,2017,35(3):152-157,162(in Chinese)薛林贵,姜金融,Famous E.城市空气微生物的监测及研究进展[J].环境工程,2017,35(3):152-157,162
[27]Zhang GB,Xue P,Hou WF,et al.The study on micro-environment of the cave affected by the visitors of the Mogao Grottoes[J].Dunhuang Research,2005(4):83-86(in Chinese)张国彬,薛平,侯文芳,等.游客流量对莫高窟洞窟内小环境的影响研究[J].敦煌研究,2005(4):83-86
[28]Saiz-jimenez C,Cuezva S,Jurado V,et al.Paleolithic art in peril:policy and science collide at Altamira Cave[J].Science,2011,334(6052):42-43
[29]Bastian F,Alabouvette C,Jurado V,et al.Impact of biocide treatments on the bacterial communities of the Lascaux Cave[J].Naturwissenschaften,2009,96(7):863-868
[30]Yan F,Ge QY,Li Q,et al.Analysis of microbial community on the surface of the historic stone and nearby rock samples in Yungang Grottoes[J].Acta Microbiologica Sinica,2012,52(5):629-636(in Chinese)颜菲,葛琴雅,李强,等.云冈石窟石质文物表面及周边岩石样品中微生物群落分析[J].微生物学报,2012,52(5):629-636
[31]Carlo ED,Chisesi R,Barresi G,et al.Fungi and bacteria in indoor cultural heritage environments:microbial-related risks for artworks and human health[J].Environment and Ecology Research,2016,4(5):257-264
[32]Rosa R,Depascale D,Cleary T,et al.Differential environmental contamination with Acinetobacter baumannii based on the anatomic source of colonization[J].American Journal of Infection Control,2014,42(7):755-757
[33]Ciferri O.Microbial degradation of paintings[J].Applied and Environmental Microbiology,1999,65(3):879-885
[34]Pepe O,Sannino L,Palomba S,et al.Heterotrophic microorganisms in deteriorated medieval wall paintings in southern Italian churches[J].Microbiological Research,2010,165(1):21-32
[35]Heyrman J,Mergaert J,Denys R,et al.The use of fatty acid methyl ester analysis(FAME)for the identification of heterotrophic bacteria present on three mural paintings showing severe damage by microorganisms[J].FEMSMicrobiology Letters,1999,181(1):55-62
[36]Gonzalez I,Laiz L,Hermosin B,et al.Bacteria isolated from rock art paintings:the case of Atlanterra shelter(south Spain)[J].Journal of Microbiological Methods,1999,36(1/2):123-127
[2]Schabereiter-Gurtner C,Saiz-Jimenez C,Pi?ar G,et al.Phylogenetic diversity of bacteria associated with Paleolithic paintings and surrounding rock walls in two Spanish caves(Llonín and La Garma)[J].FEMSMicrobiology Ecology,2004,47(2):235-247
[3]Ma YT,Zhang H,Du Y,et al.The community distribution of bacteria and fungi on ancient wall paintings of the Mogao Grottoes[J].Scientific Reports,2015,5:7752
[4]Duan YL,Wu FS,Wang WF,et al.The microbial community characteristics of ancient painted sculptures in Maijishan Grottoes,China[J].PLoS One,2017,12(7):e0179718
[5]Bastian F,Jurado V,NovákováA,et al.The microbiology of Lascaux Cave[J].Microbiology,2010,156(3):644-652
[6]Dupont J,Jacquet C,Dennetière B,et al.Invasion of the French Paleolithic painted cave of Lascaux by members of the Fusarium solani species complex[J].Mycologia,2007,99(4):526-533
[7]Portillo MC,Saiz-Jimenez C,Gonzalez JM.Molecular characterization of total and metabolically active bacterial communities of‘‘white colonizations’’in the Altamira Cave,Spain[J].Research in Microbiology,2009,160(1):41-47
[8]Schabereiter-Gurtner C,Saiz-Jimenez C,Pi?ar G,et al.Phylogenetic 16S rRNA analysis reveals the presence of complex and partly unknown bacterial communities in Tito Bustillo cave,Spain,and on its Palaeolithic paintings[J].Environmental Microbiology,2002,4(7):392-400
[9]Song LH,Song WM,Shi W,et al.Health effects of atmospheric microbiological pollution on respiratory system among children in Shanghai[J].Journal of Environment and Health,2000,17(3):135-138(in Chinese)宋凌浩,宋伟民,施玮,等.上海市大气微生物污染对儿童呼吸系统健康影响的研究[J].环境与健康杂志,2000,17(3):135-138
[10]Wang WF,Zhao LY,Pei QQ,et al.Exploration and practice of conservation of wall paintings in the museum:the case of conservation of wall paintings collected in the Gansu Museum from Tiantishan Grottoes,Wuwei[J].Sciences of Conservation and Archaeology,2015,27(4):101-112(in Chinese)汪万福,赵林毅,裴强强,等.馆藏壁画保护理论探索与实践--以甘肃省博物馆藏武威天梯山石窟壁画的保护修复为例[J].文物保护与考古科学,2015,27(4):101-112
[11]Bonazza A,de Nuntiis P,Mandrioli P,et al.Aerosol impact on cultural heritage:deterioration processes and strategies for preventive conservation[A]//Tomasi C,Fuzzi S,Kokhanovsky A.Atmospheric Aerosols:Life Cycles and Effects on Air Quality and Climate[M].Weinheim:Wiley,2016:645-670
[12]Wang WF,Ma YT,Ma X,et al.Seasonal variations of airborne bacteria in the Mogao Grottoes,Dunhuang,China[J].International Biodeterioration&Biodegradation,2010,64(4):309-315
[13]Wang WF,Ma X,Ma YT,et al.Seasonal dynamics of airborne fungi in different caves of the Mogao Grottoes,Dunhuang,China[J].International Biodeterioration&Biodegradation,2010,64(6):461-466
[14]Docampo S,Trigo MM,Recio M,et al.Fungal spore content of the atmosphere of the cave of Nerja(southern Spain):diversity and origin[J].Science of the Total Environment,2011,409(4):835-843
[15]Wu WT,He HP,Yan L,et al.The isolation and identification of bacteria in the air of Capital Museum and their significance for protection of cultural relics[J].Sciences of Conservation and Archaeology,2012,24(1):76-82(in Chinese)武望婷,何海平,闫丽,等.首都博物馆空气中细菌的分离鉴定及在文物保护中的意义[J].文物保护与考古科学,2012,24(1):76-82
[16]E J,Wu FS,Wang WF,et al.Monitoring and research on microbes in the environment of the wall paintings in No.5of the Wei and Jin Tombs[J].Dunhuang Research,2013(6):109-116(in Chinese)俄军,武发思,汪万福,等.魏晋五号壁画墓保存环境中空气微生物监测研究[J].敦煌研究,2013(6):109-116
[17]Wang WF,Ma YT,Ma X,et al.Diversity and seasonal dynamics of airborne bacteria in the Mogao Grottoes,Dunhuang,China[J].Aerobiologia,2012,28(1):27-38
[18]Tang H,Fan WQ,Wang C,et al.A dynamic study on the species and quantity of microorganisms in the micro-environment,Chongqing China Three Gorges Museum[J].Sciences of Conservation and Archaeology,2017,29(1):35-43(in Chinese)唐欢,范文奇,王春,等.重庆中国三峡博物馆小环境空气微生物种属与数量的动态研究[J].文物保护与考古科学,2017,29(1):35-43
[19]Gaüzère C,Moletta-Denat M,Blanquart H,et al.Stability of airborne microbes in the Louvre Museum over time[J].Indoor Air,2014,24(1):29-40
[20]Gaüzère C,Moletta-Denat M,Bousta F,et al.Airborne microbiology in museum through molecular approach:preliminary study[C].Indoor Air Quality,2010
[21]Ruga L,Orlandi F,Romano B,et al.The assessment of fungal bioaerosols in the crypt of St.Peter in Perugia(Italy)[J].International Biodeterioration&Biodegradation,2015,98:121-130
[22]Gaüzère C,Moletta-Denat M,Bousta F,et al.Reliable procedure for molecular analysis of airborne microflora in three indoor environments:an office and two different museum contexts[J].Clean-Soil,Air,Water,2013,41(3):226-234
[23]Anaya M,Borrego SF,Gámez E,et al.Viable fungi in the air of indoor environments of the National Archive of the Republic of Cuba[J].Aerobiologia,2016,32(3):513-527
[24]Lazaridis M,Katsivela E,Kopanakis I,et al.Indoor/outdoor particulate matter concentrations and microbial load in cultural heritage collections[J].Heritage Science,2015,3:34
[25]Tanaka D,Terada Y,Nakashima T,et al.Seasonal variations in airborne bacterial community structures at a suburban site of central Japan over a 1-year time period using PCR-DGGEmethod[J].Aerobiologia,2015,31(2):143-157
[26]Xue LG,Jiang JR,Famous E.Progress in research and monitoring of urban airborne microbes[J].Environmental Engineering,2017,35(3):152-157,162(in Chinese)薛林贵,姜金融,Famous E.城市空气微生物的监测及研究进展[J].环境工程,2017,35(3):152-157,162
[27]Zhang GB,Xue P,Hou WF,et al.The study on micro-environment of the cave affected by the visitors of the Mogao Grottoes[J].Dunhuang Research,2005(4):83-86(in Chinese)张国彬,薛平,侯文芳,等.游客流量对莫高窟洞窟内小环境的影响研究[J].敦煌研究,2005(4):83-86
[28]Saiz-jimenez C,Cuezva S,Jurado V,et al.Paleolithic art in peril:policy and science collide at Altamira Cave[J].Science,2011,334(6052):42-43
[29]Bastian F,Alabouvette C,Jurado V,et al.Impact of biocide treatments on the bacterial communities of the Lascaux Cave[J].Naturwissenschaften,2009,96(7):863-868
[30]Yan F,Ge QY,Li Q,et al.Analysis of microbial community on the surface of the historic stone and nearby rock samples in Yungang Grottoes[J].Acta Microbiologica Sinica,2012,52(5):629-636(in Chinese)颜菲,葛琴雅,李强,等.云冈石窟石质文物表面及周边岩石样品中微生物群落分析[J].微生物学报,2012,52(5):629-636
[31]Carlo ED,Chisesi R,Barresi G,et al.Fungi and bacteria in indoor cultural heritage environments:microbial-related risks for artworks and human health[J].Environment and Ecology Research,2016,4(5):257-264
[32]Rosa R,Depascale D,Cleary T,et al.Differential environmental contamination with Acinetobacter baumannii based on the anatomic source of colonization[J].American Journal of Infection Control,2014,42(7):755-757
[33]Ciferri O.Microbial degradation of paintings[J].Applied and Environmental Microbiology,1999,65(3):879-885
[34]Pepe O,Sannino L,Palomba S,et al.Heterotrophic microorganisms in deteriorated medieval wall paintings in southern Italian churches[J].Microbiological Research,2010,165(1):21-32
[35]Heyrman J,Mergaert J,Denys R,et al.The use of fatty acid methyl ester analysis(FAME)for the identification of heterotrophic bacteria present on three mural paintings showing severe damage by microorganisms[J].FEMSMicrobiology Letters,1999,181(1):55-62
[36]Gonzalez I,Laiz L,Hermosin B,et al.Bacteria isolated from rock art paintings:the case of Atlanterra shelter(south Spain)[J].Journal of Microbiological Methods,1999,36(1/2):123-127
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