麦积山石窟赋存环境中空气细菌的时空分布特征
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摘要
【目的】空气微生物沉降及污染与文化遗产的微生物退化密切相关,本文对世界文化遗产地麦积山石窟赋存环境空气中细菌浓度和群落结构的季节性变化特征进行了系统研究,为石窟环境监测预警和文物预防性保护提供依据。【方法】利用生物气溶胶采样器,在2016年春、夏、秋和冬季分别采集空气样品;基于传统培养方法获得空气中细菌浓度及纯培养菌株;通过提取基因组DNA、扩增细菌16S rRNA、测序和系统发生树等分子技术研究细菌群落时空动态变化规律;结合环境监测数据,分析影响遗产地空气细菌变化的主要因素。【结果】监测期内,空气细菌浓度在(281.20–1409.20)CFU/m3之间,最高浓度出现在MJ4处的夏季,最低浓度出现在MJO处的春季;具有明显季节性变化特征,在空间层位分布上有所差异,但不显著(P>0.05)。培养的细菌菌株经鉴定属于4个门11个属;芽孢杆菌属(Bacillus)、Paenarthrobacter、节杆菌属(Arthrobacter)、薄层菌属(Hymenobacter)和考克氏菌属(Kocuria)等为优势属。【结论】麦积山石窟空气细菌群落结构具有明显的季节性和空间分布动态变化特征;在石窟不同层位,空气中细菌群落分布与相对湿度、温度与降雨量相关;部分细菌种属如芽孢杆菌属、微球菌属(Micrococcus),为壁画及彩塑生物腐蚀的潜在病害菌;麦积山石窟及周边环境空气细菌的监测可为石窟保护和旅游开放管理提供重要参考。
[Objective] The deposition of the airborne microorganisms and air pollution were closely related to the biodeterioration of the cultural heritage. In this study, a systematic survey of seasonal variation characteristics of bacterial concentration and community structure was carried out in the surrounding atmosphere of Maijishan Grottoes, a renowned cultural heritage site in China. The results will be helpful for the environmental monitoring and precaution of cave temples and preventive protection of cultural relics. [Methods] The Bio-aerosol sampler was used for sampling in all 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 r RNA region, sequencing, and phylogenetic analysis, thereafter the bacteria community composition and distribution characteristics were clarified. Combined with environmental monitoring data, to find out the main factors which may responsible for the dynamic changes of the airborne bacteria. [Results] The concentrations of culturable bacteria were in a range from 281.2 to 1409.2 CFU/m3, the highest and the lowest concentration appears at MJ4 in summer and at MJO in spring, which varied apparently with seasonal characteristics, but no significant difference among different sites. A total of 11 different bacterial genera that affiliated to four phyla were detected in this study, among which, Bacillus, Paenarthrobacter, Arthrobacter, Hymenobacter and Kocuria were dominant genera. [Conclusion] The airborne bacterial community structures of the Maijishan grottoes show a dynamic characteristic of the seasonal variation and spatial distribution; RH, temperature and rainfall all have influenced on the airborne bacteria distribution in different layers of the Grottoes; some of the genus, such as Bacillus and Micrococcus, has potentials that could result in the biodeterioration of the ancient wall paintings and painted sculptures at this site; the monitoring of the airborne bacteria in its surrounding atmosphere of the Maijishan grottoes may contribute to the grottoes conservation and tourism management.
[Objective] The deposition of the airborne microorganisms and air pollution were closely related to the biodeterioration of the cultural heritage. In this study, a systematic survey of seasonal variation characteristics of bacterial concentration and community structure was carried out in the surrounding atmosphere of Maijishan Grottoes, a renowned cultural heritage site in China. The results will be helpful for the environmental monitoring and precaution of cave temples and preventive protection of cultural relics. [Methods] The Bio-aerosol sampler was used for sampling in all 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 r RNA region, sequencing, and phylogenetic analysis, thereafter the bacteria community composition and distribution characteristics were clarified. Combined with environmental monitoring data, to find out the main factors which may responsible for the dynamic changes of the airborne bacteria. [Results] The concentrations of culturable bacteria were in a range from 281.2 to 1409.2 CFU/m3, the highest and the lowest concentration appears at MJ4 in summer and at MJO in spring, which varied apparently with seasonal characteristics, but no significant difference among different sites. A total of 11 different bacterial genera that affiliated to four phyla were detected in this study, among which, Bacillus, Paenarthrobacter, Arthrobacter, Hymenobacter and Kocuria were dominant genera. [Conclusion] The airborne bacterial community structures of the Maijishan grottoes show a dynamic characteristic of the seasonal variation and spatial distribution; RH, temperature and rainfall all have influenced on the airborne bacteria distribution in different layers of the Grottoes; some of the genus, such as Bacillus and Micrococcus, has potentials that could result in the biodeterioration of the ancient wall paintings and painted sculptures at this site; the monitoring of the airborne bacteria in its surrounding atmosphere of the Maijishan grottoes may contribute to the grottoes conservation and tourism management.
引文
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[6]Jones AM,Harrison RM.The effects of meteorological factors on atmospheric bioaerosol concentrations-a review.Science of the Total Environment,2004,326(1/3):151-180.
[7]E J,Wu FS,Wang WF,Chen GL,Zhao LY,He DP,Xu RH.Monitoring and research on microbes in the environment of the wall paintings in No.5 of the Wei and Jin Tombs.Dunhuang Research,2013,(6):109-116.(in Chinese)俄军,武发思,汪万福,陈庚龄,赵林毅,贺东鹏,徐瑞红.魏晋五号壁画墓保存环境中空气微生物监测研究.敦煌研究,2013,(6):109-116.
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[18]Bastian F,Alabouvette C,Jurado V,Saiz-Jimenez C.Impact of biocide treatments on the bacterial communities of the Lascaux Cave.Naturwissenschaften,2009,96(7):863-868.
[19]Yan F,Ge QY,Li Q,Yu M,Zhu XD,Pan J.Analysis of microbial community on the surface of the historic stone and nearby rock samples in Yungang Grottoes.Acta Microbiologica Sinica,2012,52(5):629-636.(in Chinese)颜菲,葛琴雅,李强,于淼,朱旭东,潘皎.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.微生物学报,2012,52(5):629-636.
[20]Sanchez-Moral S,Luque L,Cuezva S,Soler V,Benavente D,Laiz L,Gonzalez JM,Saiz-Jimenez C.Deterioration of building materials in Roman catacombs:the influence of visitors.Science of the Total Environment,2005,349(1/3):260-276.
[21]Godoi RHM,Potgieter-Vermaak S,Godoi AFL,Stranger M,van Grieken R.Assessment of aerosol particles within the Rubens’House Museum in Antwerp,Belgium.X-Ray Spectrometry,2010,37(4):298-303.
[22]Duan YL,Wu FS,Wang WF,He DP,Gu JD,Feng HY,Chen T,Liu GX,An LZ.The microbial community characteristics of ancient painted sculptures in Maijishan Grottoes,China.PLoS One,2017,12(7):e0179718.
[23]Li XD,Qu JH,Zhou J,Zhang LJ,Li HF,Tian HL.Effect of sediment extract on the culturable aerobic bacterial diversity in the sediments of Taihu Lake.Microbiology China,2017,44(3):554-560.(in Chinese)李晓丹,屈建航,周佳,张璐洁,李海峰,田海龙.泥浸汁对太湖沉积物中的好氧可培养细菌多样性的影响.微生物学通报,2017,44(3):554-560.
[24]Bai XH,Wu HL,Wang HL,Zhu B.Comparison of different measurement methods of heterotrophic plate count in drinking water.Water Purification Technology,2007,26(5):65-67.(in Chinese)白晓慧,吴汉靓,王海亮,朱斌.饮用水中异养菌平板计数检测方法的比较.净水技术,2007,26(5):65-67.
[25]Tanaka D,Terada Y,Nakashima T,Sakatoku A,Nakamura S.Seasonal variations in airborne bacterial community structures at a suburban site of central Japan over a 1-year time period using PCR-DGGE method.Aerobiologia,2015,31(2):143-157.
[26]Xue LG,Jiang JR,Famous E.Progress in research and monitoring of urban airborne microbes.Environmental Engineering,2017,35(3):152-157.(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.环境工程,2017,35(3):152-157.
[27]韩明臣.城市森林保健功能指数评价研究--以北宫国家森林公园为例.中国林业科学研究院博士学位论文,2011.
[28]Porca E,Jurado V,Martin-Sanchez PM,Hermosin B,Bastian F,Alabouvette C,Saiz-Jimenez C.Aerobiology:an ecological indicator for early detection and control of fungal outbreaks in caves.Ecological Indicators,2011,11(6):1594-1598.
[29]Wang WF,Ma YT,Ma X,Wu FS,Ma XJ,An LZ,Feng HY.Diversity and seasonal dynamics of airborne bacteria in the Mogao Grottoes,Dunhuang,China.Aerobiologia,2012,28(1):27-38.
[30]Laiz L,Gonzalez JM,Saiz-Jimenez C.Microbial communities in caves:ecology,physiology,and effects on Paleolithic paintings//Koestler RJ.Art,Biology,and Conservation:Biodeterioration of Works of Art.New York:The Metropolitan Museum of Art,2003:210-215.
[31]Martin-Sanchez PM,Jurado V,Porca E,Bastian F,Lacanette D,Alabouvette C,Saiz-Jimenez C.Airborne microorganisms in Lascaux Cave(France).International Journal of Speleology,2014,43(3):295-303.
[32]Gaüzère C,Moletta-Denat M,Blanquart H,Ferreira S,Moularat S,Godon JJ,Robine E.Stability of airborne microbes in the Louvre Museum over time.Indoor Air,2014,24(1):29-40.
[33]Orial G,Bousta F,Fran?ois A,Pallot-Frossard I,Warscheid T.Managing biological activities in Lascaux:identification of microorganisms,monitoring and treatments//Coye N.Lascaux and preservation Issues in Subterranean Environments.Proceedings of the International Symposium.Paris:éditions de la Maison des sciences de l’homme,2011:219-251.
[34]Laiz L,Groth I,Gonzalez I,Saiz-Jimenez C.Microbiological study of the dripping waters in Altamira cave(Santillana del Mar,Spain).Journal of Microbiological Methods,1999,36(1/2):129-138.
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[2]Dupont J,Jacquet C,Dennetière B,Lacoste S,Bousta F,Orial G,Cruaud C,Couloux A,Roquebert MF.Invasion of the French Paleolithic painted cave of Lascaux by members of the Fusarium solani species complex.Mycologia,2007,99(4):526-533.
[3]Portillo MC,Saiz-Jimenez C,Gonzalez JM.Molecular characterization of total and metabolically active bacterial communities of“white colonizations’’in the Altamira Cave,Spain.Research in Microbiology,2009,160(1):41-47.
[4]Schabereiter-Gurtner C,Saiz-Jimenez C,Pi?ar G,Lubitz W,R?lleke S.Phylogenetic 16S rRNA analysis reveals the presence of complex and partly unknown bacterial communities in Tito Bustillo cave,Spain,and on its Palaeolithic paintings.Environmental Microbiology,2002,4(7):392-400.
[5]Schabereiter-Gurtner C,Saiz-Jimenez C,Pi?ar G,Lubitz W,R?lleke S.Phylogenetic diversity of bacteria associated with Paleolithic paintings and surrounding rock walls in two Spanish caves(Llonín and La Garma).FEMS Microbiology Ecology,2004,47(2):235-247.
[6]Jones AM,Harrison RM.The effects of meteorological factors on atmospheric bioaerosol concentrations-a review.Science of the Total Environment,2004,326(1/3):151-180.
[7]E J,Wu FS,Wang WF,Chen GL,Zhao LY,He DP,Xu RH.Monitoring and research on microbes in the environment of the wall paintings in No.5 of the Wei and Jin Tombs.Dunhuang Research,2013,(6):109-116.(in Chinese)俄军,武发思,汪万福,陈庚龄,赵林毅,贺东鹏,徐瑞红.魏晋五号壁画墓保存环境中空气微生物监测研究.敦煌研究,2013,(6):109-116.
[8]Bonazza A,de Nuntiis P,Mandrioli P,Sabbioni C.Aerosol impact on cultural heritage:deterioration processes and strategies for preventive conservation//Tomasi C,Fuzzi S,Kokhanovsky A.Atmospheric Aerosols:Life Cycles and Effects on Air Quality and Climate.Weinheim:Wiley,2017:645-670.
[9]Huang CY,Lee CC,Li FC,Ma YP,Su HJJ.The seasonal distribution of bioaerosols in municipal landfill sites:a 3-yr study.Atmospheric Environment,2002,36(27):4385-4395.
[10]Maron PA,Mougel C,Lejon DPH,Carvalho E,Bizet K,Marck G,Cubito N,Lemanceau P,Ranjard L.Temporal variability of airborne bacterial community structure in an urban area.Atmospheric Environment,2006,40(40):8074-8080.
[11]Giorgio CD,Krempff A,Guiraud H,Binder P,Tiret C,Dumenil G.Atmospheric pollution by airborne microorganisms in the city of Marseilles.Atmospheric Environment,1996,30(1):155-160.
[12]Liu LJS,Krahmer M,Fox A,Feigley CE,Featherstone A,Saraf A,Larsson L.Investigation of the concentration of bacteria and their cell envelope components in indoor air in two elementary schools.Journal of the Air&Waste Management Association,2000,50(11):1957-1967.
[13]Lugauskas A,Sveistyte L,Ulevicius V.Concentration and species diversity of airborne fungi near busy streets in Lithuanian urban areas.Annals of Agricultural and Environmental Medicine,2003,10(2):233-239.
[14]Zhang GB,Xue P,Hou WF,Guo QL.The study on micro-environment of the cave affected by the visitors of the Mogao grottoes.Dunhuang Research,2005,(4):83-86.(in Chinese)张国彬,薛平,侯文芳,郭青林.游客流量对莫高窟洞窟内小环境的影响研究.敦煌研究,2005,(4):83-86.
[15]Saiz-jimenez C,Cuezva S,Jurado V,Fernandez-Cortes A,Porca E,Benavente D,Ca?averas JC,Sanchez-Moral S.Paleolithic art in peril:policy and science collide at Altamira Cave.Science,2011,334(6052):42-43.
[16]Wang WF,Ma YT,Ma X,Wu FS,Ma XJ,An LZ,Feng HY.Seasonal variations of airborne bacteria in the Mogao Grottoes,Dunhuang,China.International Biodeterioration&Biodegradation,2010,64(4):309-315.
[17]Wang WF,Ma X,Ma YT,Mao L,Wu FS,Ma XJ,An LZ,Feng HY.Seasonal dynamics of airborne fungi in different caves of the Mogao Grottoes,Dunhuang,China.International Biodeterioration&Biodegradation,2010,64(6):461-466.
[18]Bastian F,Alabouvette C,Jurado V,Saiz-Jimenez C.Impact of biocide treatments on the bacterial communities of the Lascaux Cave.Naturwissenschaften,2009,96(7):863-868.
[19]Yan F,Ge QY,Li Q,Yu M,Zhu XD,Pan J.Analysis of microbial community on the surface of the historic stone and nearby rock samples in Yungang Grottoes.Acta Microbiologica Sinica,2012,52(5):629-636.(in Chinese)颜菲,葛琴雅,李强,于淼,朱旭东,潘皎.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.微生物学报,2012,52(5):629-636.
[20]Sanchez-Moral S,Luque L,Cuezva S,Soler V,Benavente D,Laiz L,Gonzalez JM,Saiz-Jimenez C.Deterioration of building materials in Roman catacombs:the influence of visitors.Science of the Total Environment,2005,349(1/3):260-276.
[21]Godoi RHM,Potgieter-Vermaak S,Godoi AFL,Stranger M,van Grieken R.Assessment of aerosol particles within the Rubens’House Museum in Antwerp,Belgium.X-Ray Spectrometry,2010,37(4):298-303.
[22]Duan YL,Wu FS,Wang WF,He DP,Gu JD,Feng HY,Chen T,Liu GX,An LZ.The microbial community characteristics of ancient painted sculptures in Maijishan Grottoes,China.PLoS One,2017,12(7):e0179718.
[23]Li XD,Qu JH,Zhou J,Zhang LJ,Li HF,Tian HL.Effect of sediment extract on the culturable aerobic bacterial diversity in the sediments of Taihu Lake.Microbiology China,2017,44(3):554-560.(in Chinese)李晓丹,屈建航,周佳,张璐洁,李海峰,田海龙.泥浸汁对太湖沉积物中的好氧可培养细菌多样性的影响.微生物学通报,2017,44(3):554-560.
[24]Bai XH,Wu HL,Wang HL,Zhu B.Comparison of different measurement methods of heterotrophic plate count in drinking water.Water Purification Technology,2007,26(5):65-67.(in Chinese)白晓慧,吴汉靓,王海亮,朱斌.饮用水中异养菌平板计数检测方法的比较.净水技术,2007,26(5):65-67.
[25]Tanaka D,Terada Y,Nakashima T,Sakatoku A,Nakamura S.Seasonal variations in airborne bacterial community structures at a suburban site of central Japan over a 1-year time period using PCR-DGGE method.Aerobiologia,2015,31(2):143-157.
[26]Xue LG,Jiang JR,Famous E.Progress in research and monitoring of urban airborne microbes.Environmental Engineering,2017,35(3):152-157.(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.环境工程,2017,35(3):152-157.
[27]韩明臣.城市森林保健功能指数评价研究--以北宫国家森林公园为例.中国林业科学研究院博士学位论文,2011.
[28]Porca E,Jurado V,Martin-Sanchez PM,Hermosin B,Bastian F,Alabouvette C,Saiz-Jimenez C.Aerobiology:an ecological indicator for early detection and control of fungal outbreaks in caves.Ecological Indicators,2011,11(6):1594-1598.
[29]Wang WF,Ma YT,Ma X,Wu FS,Ma XJ,An LZ,Feng HY.Diversity and seasonal dynamics of airborne bacteria in the Mogao Grottoes,Dunhuang,China.Aerobiologia,2012,28(1):27-38.
[30]Laiz L,Gonzalez JM,Saiz-Jimenez C.Microbial communities in caves:ecology,physiology,and effects on Paleolithic paintings//Koestler RJ.Art,Biology,and Conservation:Biodeterioration of Works of Art.New York:The Metropolitan Museum of Art,2003:210-215.
[31]Martin-Sanchez PM,Jurado V,Porca E,Bastian F,Lacanette D,Alabouvette C,Saiz-Jimenez C.Airborne microorganisms in Lascaux Cave(France).International Journal of Speleology,2014,43(3):295-303.
[32]Gaüzère C,Moletta-Denat M,Blanquart H,Ferreira S,Moularat S,Godon JJ,Robine E.Stability of airborne microbes in the Louvre Museum over time.Indoor Air,2014,24(1):29-40.
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