Study on the Inhibitory Effects of Plant Distillates on Microbial Communities in the lampenflora of Karst Tourist Caves

Study on the Inhibitory Effects of Plant Distillates on Microbial Communities in the lampenflora of Karst Tourist Caves | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Study on the Inhibitory Effects of Plant Distillates on Microbial Communities in the lampenflora of Karst Tourist Caves ZHIYI Xu, Shengyu Yang, Jinru Liu, Kehua Wu, MingZhong Long This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6176730/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Currently, the ecosystems of karst caves faces serious prpblems of biological weathering and environmental damage caused by lampenflora. This issue is exacerbated by artificial lighting sources and tourist activities in cave tourism development, which promote the excessive growth of these plants and result in the degradation of cave rocks and the surrounding ecological environment. Therefore, the management of lampenflora is a key task in cave ecosystem conservation. This study explores the inhibitory effects of plant distillates, specifically from mint, mugwort, and cinnamon, on lampenflora in karst caves as an eco-friendly and sustainable solution. Through experiments in Zhijin Cave, Guizhou Province, the inhibitory effects of different plant distillates at different concentrations on lampenflora communities and their microbial communities were studied. The results showed that mugwort distillate had the most significant inhibitory effect on bacterial and fungal communities, with microbial diversity and richness significantly reduced in samples treated with mugwort distillate. Mint and cinnamon distillates showed relatively weaker effects, with mint exhibiting selective inhibition on certain fungal species. Overall, plant distillates, particularly mugwort distillate, demonstrated great potential in protecting cave ecosystems, inhibiting lampenflora, and controlling associated microbial communities. The findings confirm that plant distillates can effectively suppress the growth of lampenflora, reduce microbial community richness and diversity, and thereby slow down the biological weathering process in caves. This research provides a new and sustainable solution for the ecological management in karst caves, which has high application value and environmental adaptability. Biological sciences/Ecology/Microbial ecology Earth and environmental sciences/Environmental social sciences/Sustainability Karst Caves lampenflora Ecological Restoration Cave Ecosystem Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Caves are typical representatives of karst landforms, usually formed by the gradual expansion of fissures in soluble rock masses by groundwater through dissolution and erosion processes, resulting in relatively closed and structurally stable underground spaces [ 1 ] . Since the temperature and humidity inside the cave are usually constant, these environments seem ideal. However, they are highly sensitive to external changes and human activities and exhibit low environmental carrying capacity, making them extremely vulnerable ecologically [ 2 , 3 ] . Therefore, during cave development, such as building trails, installing lighting systems and other infrastructure, the originally closed natural structure will be disrupted. The resulting dust pollution and increased air exchange between the cave interior and exterior allow the invasion of exogenous organisms such as bacteria, archaea, fungi, lichens, insects, arachnids, and arthropods, posing a serious threat to the stability of the cave ecosystem. Furthermore, if cave management is inadequate after tourism development, issues such as the rapid proliferation of lampenflora under artificial lighting and seasonal increases in tourist numbers can lead to dramatic changes in environmental parameters like temperature, humidity, carbon dioxide concentration, and negative ion concentration. These changes may cause surface pollution, discoloration, and aging of the cave's landscape, further exacerbating ecological damage [ 1 , 4 ] . These phenomena have become key environmental problems that need to be addressed in the development of cave tourism. Particularly in karst caves, the biological weathering caused by Lampenflora (referred to as "lamp flora") is especially prominent. These algae, mosses, and lichen communities typically grow within the range of artificial light sources, which provide them with suitable light and temperature conditions that promote their rapid reproduction. Depending on the intensity of light inside the cave, these lampenflora can be classified into different zones: the Chlorella and Chlorella-like algae zone, moss and lichen zone, lichen and algae zone, and fungi and algae zone [ 2 , 5 – 7 ] . The growth and reproduction of lampenflora in karst caves primarily depend on organic matter residues within the cave, such as groundwater, animal excrement, and dust, hair brought in by tourists. These plants engage in complex ecological cycles through interactions between autotrophic and heterotrophic organisms [8–13,] . At the same time, human activities, such as tourism, continually introduce additional plant and microbial species into the cave environment, making these community characteristics more prominent in tourist caves. The growth and metabolic activities of lampenflora significantly impact the surface layer of karst cave ecosystems. The organic acids secreted by these plants, along with organic matter left behind after their decay, form a porous layer on the rock surfaces within the cave, resulting in landscape corrosion and degradation of the cave's habitat for animals. Additionally, these metabolic products react with the calcium carbonate matrix, leading to the formation of precipitates that accelerate the enzyme-catalyzed dissolution of stalactites, with dissolution rates positively correlated to the activity of carbonic anhydrase (CA) [ 9 , 12 ] . Furthermore, bacteria and fungi within the lampenflora communities have a particularly pronounced effect on the rock surfaces. Bacteria degrade the rock structure through the secretion of acidic substances, while fungal hyphae penetrate the mineral structure, contributing to the physical deterioration of stalactites. The metabolic by-products of both bacteria and fungi, especially organic acids, alter the local pH, further exacerbating rock degradation [ 11 – 14 ] . The colonization rates of microorganisms are closely related to the capillarity and roughness of the substrate. Smooth substrates retain moisture for longer periods but with lower moisture content, and while the initial colonization rate may be lower, microbial survival rates tend to be higher under reduced moisture conditions. As a result, lampenflora communities typically proliferate in moist, porous areas [ 15 , 16 ] . The spatial distribution of these communities is influenced by both light intensity and variations in temperature and humidity, which further affect community composition and metabolic activities, thus amplifying disturbances to the cave's microenvironment. Although the temperature and humidity within caves remain relatively constant, light intensity and distribution have a significant impact on the growth of lampenflora. When the light intensity decreases, the thickness of the biofilm will decrease accordingly. Therefore, addressing the issue of lampenflora is a critical component of cave ecosystem conservation [ 15 , 11 , 17 ] . The control of lampenflora encompasses a variety of approaches, including physical, chemical, and emerging technologies. Physical methods, such as mechanical removal, utilize brushes or high-pressure water jets to remove the biofilms of lampenflora from rock surfaces. High-pressure water jets are particularly effective in areas with mature and structurally complex light-loving plant communities but may damage the mineral structure and promote the spread of algal spores, so they should be combined with other methods for optimal results [ 18 , 19 ] . As a non-thermal effect technology, Radiofrequency (RF) technology uses electromagnetic waves to effectively reduce biofilm coverage without altering the rock matrix. It has achieved promising results in the laboratory and is expected to be further promoted in the future [ 20 ] . Ultraviolet (UV-C) radiation, known for its strong bactericidal ability, is widely used in controlling lampenflora, while the new far UV-C technology, by controlling the wavelength, offers a balance between safety and efficiency, providing new possibilities for management [ 21 – 23 ] .Chemical methods primarily involve the use of quaternary ammonium salts, hydrogen peroxide, and chlorine-containing compounds to suppress or remove lampenflora. Benzalkonium chloride, while effective in bleaching, is difficult to degrade and may harm cave ecosystems [ 24 – 26 ] . Hydrogen peroxide is relatively eco-friendly and provides significant removal effects but may corrode calcite and lead to the recurrence of lampenflora [ 27 – 29 ] . Chlorine-based compounds, such as chlorine dioxide, have rapid decolorization capabilities but may have potential negative effects on cave-dwelling animals and minerals [ 24 , 25 ] . To address the limitations of traditional methods, combined physical and chemical approaches have become effective strategies in recent years. After using chemical agents to remove plants, high-pressure water jets are employed to clear any remaining residues, overcoming the drawbacks of single-method approaches.Moreover, plant extracts have emerged as an eco-friendly and sustainable control technology in recent years. Extracts from plants such as Artemisia annua, mugwort, and mint can adsorb nitrogen and phosphorus, reducing the nutrients required for biofilm growth. Active compounds in clove and cinnamon extracts can disrupt the cell membranes of lampenflora, effectively inhibiting the growth of algae and mosses [ 29 – 31 ] . Compared with traditional methods, plant extracts offer plant distillates have the advantages of high environmental adaptability and little impact on cave ecology, providing a green and sustainable solution for the management of karst cave lampenflora [ 32 – 33 ] . In the context of biological weathering and environmental degradation caused by the excessive growth of lampenflora in karst caves, the scientific management of lampenflora has become a critical aspect of cave conservation. Plant extracts, as an emerging biological control method, offer significant advantages due to their excellent antibacterial properties, simple preparation process, low cost, and environmentally friendly characteristics. The natural compounds in these extracts can effectively inhibit the growth of lampenflora while avoiding secondary damage to cave ecosystems. According to previous studies, leaf extracts from cinnamon (Cinnamomum verum), mint (Mentha spp.), and mugwort (Artemisia argyi) have shown promising inhibitory effects [ 34 – 36 ] . Therefore, this study selected the distillation extracts of these three plants to systematically explore their potential applications potential in the removal of lampenflora, in order to provide a scientific evidence for the efficient management of lampenflora in karst caves and proposing feasible strategies for protecting cave ecosystems. 2. Materials and Methods 2.1 Study Area The study was conducted in Zhijin Cave, located in Zhijin County, Guizhou Province, China. This area is a representative of karst landforms, with a long geological history and rich natural heritage. Due to the severe degradation caused by the growth of lampenflora, certain rocks in the cave have exhibited noticeable aesthetic deterioration and structural damage. The coverage of lampenflora not only affects the visual appeal of the cave but also accelerates the surface erosion of limestone, threatening the long-term conservation of the cave. 2.2 In situ Experiment Experimental plots were established sequentially in the selected area, with each plot measuring 27×27 cm and showing relatively complete coverage of lampenflora. Each plot was divided into a 3×3 grid, where three different concentrations of plant distillation extracts were applied. We monitored the color changes of the plants at baseline (untreated), and then at 1 day, 7 days, and 30 days after treatment, using a portable precision colorimeter. The air temperature and humidity inside the cave were recorded using a hygrometer, while light intensity was measured with a light meter. The substrate pH value was tested using a PHS-25 pH meter. Since previous experiments showed that anhydrous ethanol had no significant effect on the color change of moss, no anhydrous ethanol control group was included in this study [ 32 ] . 2.3 Plant Sample Processing and Distillation Extract Preparation In the experiment, the three plant samples purchased were first processed. The plant samples were washed with distilled water to remove surface dust and particles. After washing, the samples were air-dried naturally at room temperature (25°C), then wrapped in aluminum foil and dried in an oven at 60°C for 3 hours. Subsequently, the dried plant samples were ground into powder and sifted through a 40-mesh sieve (380 µm) to ensure sample uniformity. The plant powder was divided into three different amounts (5 g, 10 g, and 15 g), and each amount was mixed with 100 mL of anhydrous ethanol (99.7%) and heated to 78°C for distillation. The resulting clear plant distillation extracts were collected. Finally, the collected distillation extracts were stored in a refrigerator at 4°C to maintain their activity. 2.4 Microbial Sampling Three representative sampling sites were selected in Zhijin Cave for microbial sampling. During the sampling process, sterile sampling bags and scrapers were used to collect surface microbial samples from different areas, including the cave walls, ceiling, and floor. After collection, all samples were immediately stored in a refrigerated box at 4°C and transported to the laboratory for DNA high-throughput sequencing analysis within 24 hours. For the experimental samples, the control points without treatment were labeled as Y points (including A1_1, A2_1, A3_1). The points treated with mint distillation extract were labeled as B points (including A1_3, A2_3, A3_3). The points treated with mugwort distillation extract were labeled as A points (including A1_4, A2_4, A3_4). The points treated with cinnamon distillation extract were labeled as R points (including A1_2, A2_2, A3_2). 2.5 Microbial Sequencing under Different Coverage Environments 2.5.1 DNA Extraction Microbial community genomic DNA was extracted according to the instructions provided by the E.Z.N.A.® Soil DNA Kit (Omega Bio-tek, Norcross, GA, U.S.). The quality of the extracted genomic DNA was assessed using 1% agarose gel electrophoresis. DNA concentration and purity were determined using the FastDNA® Spin Kit for Soil (MP Biomedicals, U.S.). 2.5.2 PCR Amplification and Sequencing Library Construction The bacterial 16S rRNA gene region (A1_1-A3_3) was amplified using the primers 338F (5’-ACTCCTACGGGAGGCAGCAG-3’) and 806R (5’-GGACTACHVGGGTWTCTAAT-3’). The PCR reaction system included: 4 µL of 5× FastPfu buffer, 2 µL of 2.5 mM dNTPs, 0.8 µL of upstream primer (5 µM), 0.8 µL of downstream primer (5 µM), 0.4 µL of FastPfu DNA polymerase, 0.2 µL of BSA, 10 ng of template DNA, and the final volume adjusted to 20 µL. The fungal ITS rRNA gene ITS1 region was amplified using primers ITS1F (5’-CTTGGTCATTTAGAGGAAGTAA-3’) and ITS2R (5’-GCTGCGTTCTTCATCGATGC-3’). The PCR reaction system included: 2 µL of 10× rTaq buffer, 2 µL of 2.5 mM dNTPs, 0.8 µL of upstream primer (5 µM), 0.8 µL of downstream primer (5 µM), 0.2 µL of rTaq DNA polymerase, 0.2 µL of BSA, 10 ng of template DNA, and the final volume adjusted to 20 µL. The amplification program was as follows: denaturation at 95°C for 3 minutes, followed by 27 cycles (35 cycles for fungi) of 95°C for 30 seconds, 55°C for 30 seconds, and 72°C for 45 seconds, with a final extension at 72°C for 10 minutes, and stored at 4°C (PCR machine: ABI GeneAmp® 9700).The purified PCR products were then used for library construction with the NEXTFLEX Rapid DNA-Seq Kit: (1) adapter ligation, (2) magnetic bead selection to remove adapter-dimer fragments, (3) PCR amplification for library enrichment, (4) magnetic bead recovery of the PCR product to obtain the final library. Sequencing was performed on the Illumina Nextseq2000 platform (Shanghai Meiji Biotechnology Co., Ltd.). 2.5.3 Sequencing Data Processing The paired-end raw sequencing reads were quality-controlled using fastp software, and merged using FLASH software:(1) Base calls with quality scores below 20 at the tail of the reads were filtered. A 50 bp sliding window was used, and if the average quality score in the window was below 20, the bases at the end of the window were trimmed. Reads shorter than 50 bp and reads containing N bases were also removed after quality control.(2) Paired-end reads were merged based on their overlap relationship, with a minimum overlap length of 10 bp.(3) The maximum mismatch ratio allowed in the overlap region of the merged sequence was set to 0.2, and sequences that did not meet this threshold were filtered out.(4) The barcode and primer at both ends of the sequence were used to distinguish samples and adjust the sequence direction. No mismatches were allowed in the barcode, while a maximum of 2 mismatches were allowed in the primer region.To minimize the impact of sequencing depth on subsequent Alpha and Beta diversity analyses, the number of sequences in all samples was rarefied, and diversity, community composition, and other analyses were performed based on the rarefied OTU (Operational Taxonomic Unit) data. 2.5.4 DATA AVAILABILITY STATEMENT The raw data have been submitted to NCBI Sequence Read Archive (SRA) under Bioproject accession PRJNA1222770. 3. Experimental Results 3.1 Microbial Community α-Diversity Analysis and the Inhibitory Effects of Plant Distillates Microbial α-diversity was chosen to evaluate the differences in microbial communities across the three different coverage environments. The Chao index and Shannon index were used to represent the bacterial diversity, respectively. By analyzing the Chao and Shannon indices of microorganisms in the biofilms of cave lampenflora, the effects of different removal agents on the richness and diversity of microbial communities can be clearly observed. Based on both the Chao and Shannon indices, bacterial richness and diversity underwent significant changes after applying different removers. As shown in the figure, the Y point exhibited higher richness and diversity. However, after using the removers, particularly B and A, both the Chao and Shannon indices showed a marked decrease, indicating the most significant reduction in bacterial richness at point A. In comparison, the treatment effect at point R was milder, suggesting that certain bacterial phyla might exhibit tolerance to cinnamon distillate, even showing adaptive reproduction in the removal environment. In contrast to bacteria, the Chao and Shannon indices of fungi were significantly lower, indicating that the richness and diversity of fungi were relatively limited. After applying the removers, particularly at point A, the Chao and Shannon indices of fungi significantly decreased, showing that the wormwood distillate had the strongest inhibitory effect on fungi. In contrast, the inhibitory effects of mint and cinnamon distillates were relatively weaker, with mint distillate potentially exhibiting selective inhibition of certain fungal genera. These results suggest that the three plant distillates demonstrated varying efficacy in removing lampenflora and their associated microorganisms. Among them, wormwood distillate exhibited the most significant antibacterial and antifungal effects, making it the most suitable candidate for microbial management in caves. 3.2 Disruption of Microbial Community Structure in Karst Caves by Plant Distillates and Heatmap Analysis Heatmap analysis was conducted to investigate the impact of plant distillates on the bacterial and fungal communities in Karst caves, revealing the patterns and characteristics of microbial community changes under different treatment conditions. The heatmap of the bacterial community showed that the untreated group (Y) formed an independent cluster, indicating that the bacterial community structure remained unaffected by external interventions. In contrast, the groups treated with plant distillates (R, B, A) clustered together, demonstrating a significant impact of the reagents on the bacterial community structure. Specifically, the use of cinnamon distillate (R) reduced the abundance of genera such as norank_f__Spirosomaceae , Cellvibrio , and Paenibacillus , while increasing the abundance of Pedomicrobium and Leptolyngbya_ANT.L52.2 . The use of mugwort distillate (A) significantly decreased the abundance of Scytonema_PCC-7110 and norank_f__A4b , while increasing the abundance of genera such as Acinetobacter and Pseudomonas . The mint distillate (B) reduced the abundance of Hyphomicrobium and Scytonema_PCC-7110 , while significantly increasing the abundance of Pseudomonas and Massilia . These changes indicate that plant distillates can effectively disrupt the composition and metabolic function of bacterial communities in environments covered by light-sensitive plants. The heatmap of the fungal community showed a more pronounced trend of changes. The mugwort distillate (A) treatment group formed a distinct cluster, indicating a significant impact on the fungal community, while the untreated group (Y) clustered together with the cinnamon (R) and mint (B) treatment groups, reflecting similar changes in fungal communities induced by these treatments. Specifically, the mugwort distillate (A) significantly reduced the abundance of genera such as Entoloma , Trichoderma , and Penicillium , while increasing the abundance of genera like Furcasterigmium , Mortierella , and Plectosphaerella . The mint distillate (B) decreased the abundance of Leptobacillium and Ascotricha , but significantly increased the abundance of Verticillium and Brunneochlamydosporium . The cinnamon distillate (R) reduced the abundance of Rozellomycota_gen_Incertae_sedis and Trichoderma , while increasing the abundance of Metapochonia and Simplicillium . Overall, the fungal community exhibited a more sensitive response to the plant distillates, resulting in significant adjustments in community structure. Overall, the surface microenvironment of the karst cave’s lampenflora and the biofilm-covered areas formed a short-term microbial community succession process. The use of plant distillates altered the abundance and structure of microorganisms, with bacteria showing a more significant response to cinnamon and mint distillates, while the fungal community underwent a marked adjustment under mugwort distillate treatment. The results suggest that plant distillates can effectively interfere with the microbial community dynamics in light plant-covered environments, particularly exhibiting the most significant inhibitory effect on fungi. 3.3 Regulatory Effect of Plant Distillates on the Microbial and Fungal Community Structure in Karst Caves Before the plant distillate treatments (as shown in Figures A and B), the microbial community in the karst cave was dominated by bacteria. The phylum Proteobacteria (47.0%) was the absolute dominant group, while Cyanobacteria (10.7%) and Chloroflexi (9.2%) also exhibited relatively high abundance. The presence of cyanobacteria indicates that, under the light plant-covered environment, photosynthetic microorganisms occupy a certain proportion of the cave ecosystem. However, these microbial groups may accelerate rock weathering through photosynthesis, leading to carbonate dissolution and negatively impacting the structural integrity of cave walls [36] . In the fungal community, Ascomycota (60.9%) was the dominant phylum, followed by Basidiomycota (15.3%) and Rozellomycota (14.6%). After the plant distillate treatments, Figure A shows that the bacterial community composition underwent significant changes. Following cinnamon distillate (R) treatment, the abundance of Proteobacteria increased to 63.3%, while the abundance of Cyanobacteria significantly decreased to below the detection limit, indicating that cinnamon distillate effectively inhibited the photosynthetic microbial group. Additionally, the abundance of Actinobacteriota increased to 11.0%, suggesting an enrichment of microorganisms involved in organic matter decomposition.In contrast, both peppermint distillate (B) and mugwort distillate (A) treatments led to bacterial communities with higher diversity. Notably, after mugwort distillate treatment, the abundance of Actinobacteriota and Firmicutes reached 13.2% and 6.5%, respectively, indicating a significant promotion of microorganisms responsible for the breakdown of complex organic matter. Figure B illustrates the changes in fungal communities after treatment with plant distillates. After cinnamon distillate treatment, the abundance of Ascomycota significantly increased to 91.4%, while the abundance of Rozellomycota decreased to 0.4%, indicating a strong inhibitory effect of cinnamon distillate on parasitic fungi.After treatment with peppermint distillate and mugwort distillate, the abundance of Basidiomycota increased to 8.6% and 31.8%, respectively. Mugwort distillate also significantly increased the abundance of Mortierellomycota to 12.9%, demonstrating its promoting effect on the microbial group responsible for decomposing organic matter. Figures A and B visually demonstrate the significant regulatory effects of plant distillates on the structure of microbial and fungal communities. Cinnamon distillate exhibits a strong inhibitory effect, significantly reducing the abundance of harmful microbial groups such as Cyanobacteria and Rozellomycota . In contrast, peppermint and mugwort distillates show better community diversity maintenance, while also promoting the enrichment of functional microbial groups such as Actinobacteriota and Basidiomycota . 4. Spectral Characteristics Analysis of Plant Distillates and Their Mechanisms for Inhibiting Lampenflora in Karst Tourist Caves By analyzing the relationship between absorbance peaks and concentrations of mugwort, peppermint, and cinnamon distillates using ultraviolet spectrophotometry, we can not only gain a more detailed understanding of the active components responsible for removing lampenflora in karst tourist caves but also infer their activity variations and underlying mechanisms at different concentrations. The absorbance peak at 208 nm is primarily attributed to organic acids (such as benzoic acid), phenolic compounds, and flavonoids present in mugwort. As the concentration of mugwort increases, the concentration of these active components also rises, thereby enhancing the absorption of ultraviolet light and increasing mugwort's activity in plant removal. This phenomenon suggests that the natural chemical constituents of mugwort exhibit stronger UV absorption at higher concentrations, potentially providing effective support for its plant inhibitory effects. The figure below shows the absorption spectra of peppermint distillate at different concentrations within the wavelength range of 200 nm to 400 nm. The absorbance peaks of peppermint primarily occur at wavelengths of 207 nm and 229 nm. As the concentration increases, the absorbance at these two wavelengths gradually increases, demonstrating a positive correlation between concentration and absorbance peak values.At these wavelengths, the main active components in peppermint, such as menthol and peppermint oil, exhibit significant UV absorbance characteristics.Menthol and peppermint oil, as volatile organic compounds, possess strong UV absorption capacity, particularly at the 207 nm and 229 nm wavelengths.With increasing concentration, the concentration of these active components rises, further enhancing peppermint's UV light absorption ability at these two wavelengths.Furthermore, the active components in peppermint not only effectively absorb UV light but may also exert inhibitory effects on plants by releasing volatile chemicals. This effect becomes more pronounced as the concentration of peppermint increases. The figure below shows the absorption spectra of cinnamon distillate at different concentrations within the wavelength range of 200 nm to 400 nm. The absorbance peaks of cinnamon occur at 207 nm and 286 nm, with a more pronounced change in absorbance observed at 286 nm.As the concentration increases, the absorbance peak values at both wavelengths show a significant increase, with the increase in absorbance being positively correlated with concentration.This indicates that the active components in cinnamon, such as cinnamic acid, cinnamaldehyde, and coumarin, exhibit more pronounced UV absorption characteristics as the concentration increases.These components in cinnamon not only have strong antioxidant, antimicrobial, and plant growth-regulating effects, but as UV absorption increases, they may enhance the release of active components, improve biological activity, and produce stronger inhibitory effects during plant removal. 4. Discussion This study explores the potential of plant distillates in managing lampenflora in karst caves, with a particular focus on their inhibitory effects on microbial communities associated with lampenflora.The results show that the distillates of peppermint, mugwort, and cinnamon effectively inhibit the growth of lampenflora in karst caves to varying extents and significantly impact the diversity and structure of microbial communities. First, plant distillates have a significant effect on the α-diversity of microbial communities.α-diversity analysis revealed that mugwort distillate exhibited the most significant decrease in bacterial and fungal diversity, particularly in the Chao and Shannon indices.Mugwort distillate exhibited the strongest inhibitory effects on microbial communities, demonstrating potent antibacterial and antifungal activity. Further spectral analysis revealed strong absorbance signals in the 290-310 nm range for mugwort distillate. This phenomenon may be closely related to the presence of flavonoids, phenolic compounds, and certain organic acids in mugwort.Flavonoids and phenolic compounds are widely distributed in plants and are known for their significant antimicrobial, antioxidant, and UV shielding properties.Under UV light exposure, these components can effectively absorb UV light, reducing the risk of harmful UV radiation to cells. At the same time, through interactions with the cell membrane, they interfere with normal cellular functions, thereby affecting microbial growth and metabolism.Flavonoids inhibit microbial growth by suppressing photosynthesis, disrupting the integrity of cell membranes, or interfering with key metabolic pathways.Therefore, these chemical components in mugwort distillate may be the main reason for its strong inhibitory effect on microbial community diversity. The inhibitory effects of peppermint and cinnamon distillates are relatively mild. Peppermint distillate may exert inhibitory effects through its volatile components, showing a certain influence on microbial communities, especially at higher concentrations.The inhibitory effect of cinnamon distillate is more complex. Its spectral characteristics show a prominent absorbance peak in the 250-290 nm range, especially within the 280-300 nm range.The absorbance characteristics in this range may be related to the presence of coumarin compounds in cinnamon, which have been shown to possess some antimicrobial properties.Further analysis of the inhibitory mechanism of cinnamon distillate may help to better understand its impact on microbial communities. Mugwort distillate, due to its rich content of flavonoids and phenolic compounds, exhibits a strong antimicrobial effect.Peppermint and cinnamon distillates exert inhibitory effects through other components. Although their effects are not as pronounced as that of mugwort, they still impact the microbial community. 5. Conclusion The results show that all three distillates (mint, Artemisia, and cinnamon) inhibited cave plant growth at certain concentrations, with Artemisia distillate having the strongest effect on both plants and microbes. It reduced plant growth and significantly decreased microbial diversity, particularly in bacteria and fungi, highlighting its antimicrobial properties. Mint and cinnamon also inhibited growth, with mint selectively affecting certain fungi. Through the analysis of experimental data, it is evident that plant distillate, as a green and environmentally friendly inhibitor, possess considerable application potential. They can not only effectively inhibit the excessive growth of light plants, but also reduce the richness and diversity of microbial communities, thereby slowing down the biological weathering process in karst caves. It provides a sustainable solution for the protection and restoration of cave environment. In summary, the experimental results of this study validate the effectiveness of plant distillates in the management of light plants in karst caves. Moreover, this method demonstrates good environmental adaptability and can reduce negative impacts on the cave ecosystem, making it a promising approach for further promotion and application. Future research could explore the combined effects of different plant distillates under various environmental conditions, optimize management strategies, and assess their long-term effects and sustainability. Declarations Author Contribution Zhiyi Xu: Investigation, Software, Writing - Original Draft.Mingzhong Long: Project Administration, Funding Acquisition, Conceptualization, Methodology, Supervision, Writing - Original Draft, Writing - Review & Editing.Shengyu Yang: Data Curation, Methodology.Jinru Liu: Formal Analysis, Resources.Kehua Wu: Funding Acquisition, Investigation, Methodology. Acknowledgments This work was supported by the Basic Research Program of Guizhou Province(Qiankehe Jichu ZK [2023] Yiban 147) ; the Research Fund of Guizhou Minzu University (GZMUZK[2024]QD64). Data Availability Please reference PRJNA1222770 in your publication. This BioProject accession number is provided instead of SRP and should be used in your publication as it will allow better searching in Entrez.Accession to cite for these SRA data: PRJNA1222770Temporary Submission ID: SUB15085079Release date: 2025-02-12 References Katja Sterflinger, Guadalupe Piñar.Microbial deterioration of cultural heritage and worksof art — tilting at windmills?Appl Microbiol Biotechnol,2013,97:9637–9646 Nataša Nikolić, Gordana Subakov Simić, Igor Golić,Slađana Popović.The effects of biocides on the growth of aerophytic green algae (Chlorella sp.) isolated from a cave environment[J].Arch Biol Sci. 2021;73(3):341-351 Zhu Xiaoyan, Zhang Meiliang. Study on Cave Environmental Factors in Karst Cave Tourism Activities [J]. China Karst, 2020, 39(03): 426-431. Valme Jurado , Yolanda del Rosal , Jose Luis Gonzalez-Pimentel ,et al.Biological Control of Phototrophic Biofilms in a ShowCave: The Case of Nerja Cave[J].Applied sciences,2020, 10, 3448; doi:10.3390/app10103448 T.Aley. Tourist Caves: Algae and Lampenflora[C]//John.Gunn, Encyclopaedia of Caves and Karst Science. New York:Fitzroy Dearborn，2004:733-734. Esteban R P .Study and remediation of environmental problems caused due to the growth of algae in speleothems of calcareous caves adapted for tourism- a case of success in Spain [J].Journal of Environmental Geology,2018,02(01): Nataša Nikolić,C, Nikola Zarubica , Bojan Gavrilović,et al.LAMPENFLORA AND THE ENTRANCE BIOFILM IN TWO SHOW CAVES:COMPARISON OF MICROBIAL COMMUNITY,ENVIRONMENTAL,AND BIOFILM PARAMETERS.[J]Journal of Cave and Karst Studies, v. 82, no. 2, p. 69-81. DOI:10.4311/2018EX0124 Chen Weihai, Deng Yadong, Tang Li, et al. Study on the Sustainable Use of Tourism Caves in Guilin [J]. Guangxi Science, 2018, 25(05): 579-589. DOI: 10.13656/j.cnki.gxkx.20181106.003Slađana Popović, Marija Pećić ,Gordana Subakov Simić.Exploring Lampenflora of ResavskaCave, Serbia.Biology and life sciences forum.2022, 15, 33. https://doi.org/10.3390/IECD2022-12425 Slađana Popović, Marija Pećić ,Gordana Subakov Simić.Exploring Lampenflora of ResavskaCave, Serbia.Biology and life sciences forum[J].2022, 15, 33. https://doi.org/10.3390/IECD2022-12425 Zhang Kaiyan, Li Tongjian, Zhang Xianqiang, et al. The Carbonic Anhydrase of Three Lithophytic Moss Species and Its Erosion Effect on Limestone [J]. China Karst, 2017, 36(04): 441-446. Jake,Robin ,Jacob J , et al.Lampenflora in a Show Cave in the Great Basin Is Distinct from Communities on Naturally Lit Rock Surfaces in Nearby Wild Caves[J].Microorganisms,2021, Esteban R P .Study and remediation of environmental problems caused due to the growth of algae in speleothems of calcareous caves adapted for tourism- a case of success in Spain [J].Journal of Environmental Geology,2018,02(01): Hernández-Mariné M. & Roldán M., Adherence of hormogonia to substrata is mediated by polysaccharides produced by necridic cells. Algological Studies,2005,117:239-249. Mulec J ,Kosi G ,Vrhovsek D .ALGAE PROMOTE GROWTH OF STALAGMITES AND STALACTITES IN KARST CAVES (SKOCJANSKE JAME, SLOVENIA)[J].Carbonates And Evaporites,2007,22(1):6-9. Miller A.Z., Dionísio A., Laiz L., Macedo M.F. & Saiz-Jiménez C., The influence of inherent properties of building limestones on their bioreceptivity to phototrophic microorganisms. Annals of Microbiology,59: 705-713. https://doi.org/10.1007/BF03179212 David G . Caves:Processes, Development and Management[M]. Blackwell Publishing Ltd.: 1996-11-17. DOI:10.1002/9781444313680. Hernández-Mariné M ,Roldán M .Adherence of hormogonia to substrata is mediated by polysaccharides produced by necridic cells[J].Algological Studies/Archiv für Hydrobiologie, Supplement Volumes,2005,117239-249. P C ,P C ,A G , et al. Biofilms on tuff stones at historical sites: identification and removal by nonthermal effects of radiofrequencies. [J]. Microbial ecology, 2013, 66 (3): 659-68. Stéphane P ,Olympe E ,Battle K , et al.UV-C as an efficient means to combat biofilm formation in show caves: evidence from the La Glacière Cave (France) and laboratory experiments.[J].Environmental science and pollution research international,2017,24(31):24611-24623. Thomas L. Kieft, Devyn Del Curto, Zoë Havlena,et al.Potential for Mitigation of Cave Lampenflora Using BenzalkoniumChloride or UV-C.Geoheritage (2023) 15:68https://doi.org/10.1007/s12371-023-00839-4 M. Buonanno, B. Ponnaiya, D. Welch, M. Stanislauskas, G. Randers-Pehrson,L. Smilenov, F.D. Lowy, D.M. Owens, D.J. Brenner, Germicidal efficacy andmammalian skin safety of 222-nm UV light, Radiat. Res. 187 (4) (2017) 493–501,https://doi.org/10.1667/rr0010cc.1. Esteban R P . Study and remediation of environmental problems caused due to the growth of algae in speleothems of calcareous caves adapted for tourism- a case of success in Spain [J]. Journal of Environmental Geology, 2018, 02 (01): Spain Rosangela A ,Daniela B ,Beatriz C , et al.A multidisciplinary approach to the comparison of three contrasting treatments on both lampenflora community and underlying rock surface.[J].Biofouling,2023,39(2):11-14. Stéphane P ,Thomas M ,Faisl B , et al.Bleaching of biofilm-forming algae induced by UV-C treatment: a preliminary study on chlorophyll degradation and its optimization for an application on cultural heritage.[J].Environmental science and pollution research international,2018,25(14):14097-14105. Esteban R P . Study and remediation of environmental problems caused due to the growth of algae in othems of calcareous caves adapted for tourism- a case of success in Spain [J]. Journal of Environmental ogy, 2018, 02 (01): Spain Faimon J ,Štelcl J ,Kubešová S , et al.Environmentally acceptable effect of hydrogen peroxide on cave “lamp-flora”, calcite speleothems and limestones[J].Environmental Pollution,2003,122(3):417-422. Natasa N ,Subakov G S ,Igor G , et al.The effects of biocides on the growth of aerophytic green algae (Chlorella sp.) isolated from a cave environment[J].ARCHIVES OF BIOLOGICAL SCIENCES,2021,73(3):341-351. He Huiyan. Study on the Inhibition of Algal Cell Growth, Metabolic Activity, and Photosynthesis by Hydrogen Peroxide [D]. Xi'an University of Architecture and Technology, 2020. DOI: 10.27393/d.cnki.gxazu.2020.001237. Duc Anh Trinh, Quan Hong Trinh, Ngoc Tran,et al.Eco-friendly Remediation of Lamp enflora on Sp Eleothems in Tropical Karst Caves.Journal of Cave and Karst Studies, v. 80, no. 1, p. 1-12 DOI: 10.4311/2017ES0101 Addesso R ,Bellino A ,D'Angeli M I , et al.Vermiculations from karst caves: The case of Pertosa-Auletta system (Italy)[J].Catena,2019,182104178-104178. Meng Hui, Xu Yong. GC-MS Analysis of Volatile Oil Components in Fresh Artemisia argyi Leaves from Shanghai [J]. Journal of Pharmaceutical Practice, 2009, 05 - 0362 - 03. Wu Fan, Jia Ruhan. Research Progress on the Pharmacological Effects of Cinnamon Extract [J]. Herald of Medicine, 2012, 31(7): 882-885. Zheng Yao, Yang Xiaoxi, Qian Xinyu, Zhu Yuejie, Chen Jiachang, Xu Pao. Study on the Water Purification and Antibacterial Effects of Chinese Herbal Medicines. Environmental Science and Technology, 43(1), 222-228. DOI: 10.19672/j.cnki.1003-6504.2020.01.033. Lin Jiayu, Li Bangjiang, Cheng Cai, et al. Current Research on the Biological Weathering Prevention and Control of Stone Buildings [J]. Journal of Applied Ecology, 2021, 32(08): 3023-3030. DOI: 10.13287/j.1001-9332.202108.039 Yao Yongfang, Shi Lin, Tan Caideng. Study on the Extraction of Antibacterial Substances from Artemisia argyi [J]. Food Science and Technology, 2011, 36(11). DOI: 10.13684/j.cnki.spkj.2011.11.004 Yang Cuiyun, An Xin, Wan Jingqiong, et al. Food Science and Technology [J], 2021, 46(1). DOI: 10.13684/j.cnki.spkj.2021.01.030 Li Ping, Shi Chuntao, Shu Ting, Shen Xiaoxia. Comparison of Antibacterial Activity of Cinnamon Oil Extracted by Three Different Methods [J]. Preservation and Processing, 2018, 18(2): 31-38. DOI: 10.3969/j.issn.1009-6221.2018.02.006 Faimon J. Environmentally acceptable effect of hydrogen peroxide on cave "lampenflora". Calcite speleothems and limestones. Environmental Pollution, Elsevier Science Ltd. 2003:417-22. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6176730","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":439008944,"identity":"f71fa35c-51b4-4653-a0bd-0e158c9ffc0c","order_by":0,"name":"ZHIYI Xu","email":"","orcid":"","institution":"guizhou","correspondingAuthor":false,"prefix":"","firstName":"ZHIYI","middleName":"","lastName":"Xu","suffix":""},{"id":439008945,"identity":"4bc5aed8-a466-44ee-b3ed-761f33e15db5","order_by":1,"name":"Shengyu Yang","email":"","orcid":"","institution":"guizhou","correspondingAuthor":false,"prefix":"","firstName":"Shengyu","middleName":"","lastName":"Yang","suffix":""},{"id":439008946,"identity":"265f6a91-a85f-4647-936a-0605e8043be7","order_by":2,"name":"Jinru Liu","email":"","orcid":"","institution":"guizhou","correspondingAuthor":false,"prefix":"","firstName":"Jinru","middleName":"","lastName":"Liu","suffix":""},{"id":439008947,"identity":"bffab8f0-a982-4118-825d-e3671e00519b","order_by":3,"name":"Kehua Wu","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Kehua","middleName":"","lastName":"Wu","suffix":""},{"id":439008948,"identity":"47237553-f910-42cd-8df4-0d48b5f54943","order_by":4,"name":"MingZhong Long","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA70lEQVRIiWNgGAWjYBACPiidwMbABmbIsbG3H8CrhQ1Ni4ExH8+ZBOK0QJkGifMkHAzwa2HvPfzqxh+GPD72tsTPFX/+pLdJAPX/qNiGWwvPuTTrHB6GYjaeY4clz7YZ5LZJNx5g7DlzG7cWiRwz4xwJhsQ2ifQGycYGoBaZAwnMjG2EtBiAtTT/bPhjkM4mkWBASIvx45wEkJa0Y5INbAYJhLXwnDFjzjkA9kuaZWObsWEbMJAP4vMLP3uP8eccYIjJt7cZ32z4Iycv395+8MGPCtxawG5jYPiPKnQAn3ogYP5AQMEoGAWjYBSMdAAA0MRPMdKYxpUAAAAASUVORK5CYII=","orcid":"","institution":"guizhou","correspondingAuthor":true,"prefix":"","firstName":"MingZhong","middleName":"","lastName":"Long","suffix":""}],"badges":[],"createdAt":"2025-03-07 09:08:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6176730/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6176730/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":80026779,"identity":"58e36faa-2d18-4bc6-b5a7-426e54015a8e","added_by":"auto","created_at":"2025-04-07 06:33:20","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":411453,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSampling locations within Zhijin Cave, Guizhou, China, showing the lampenflora at eight selected sites.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/05702c1383ca33e699827006.png"},{"id":80026778,"identity":"17e22cce-68aa-45c1-be43-f80657cd2cfc","added_by":"auto","created_at":"2025-04-07 06:33:20","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":133913,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAlpha diversity of fungi in the karst cave\u003c/strong\u003e.\u003cbr\u003e\n \u003cstrong\u003e(A, B)\u003c/strong\u003e Chao index analysis of fungal alpha diversity, showing the richness and species estimation across the sampling points.\u003cbr\u003e\n \u003cstrong\u003e(C, D)\u003c/strong\u003e Shannon index analysis of fungal alpha diversity, reflecting the evenness and diversity of fungal communities in the cave.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/21beef4940b62cb2b60ad090.png"},{"id":80028632,"identity":"96c2d742-a2bc-4d16-8748-34a1590d540a","added_by":"auto","created_at":"2025-04-07 06:57:20","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":155473,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAlpha diversity of bacteria in the karst cave\u003c/strong\u003e.\u003cbr\u003e\n \u003cstrong\u003e(A, B)\u003c/strong\u003e Chao index analysis of bacterial alpha diversity, showing the richness and species estimation across the sampling points.\u003cbr\u003e\n \u003cstrong\u003e(C, D)\u003c/strong\u003e Shannon index analysis of bacterial alpha diversity, reflecting the evenness and diversity of bacterial communities in the cave.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/2004b45e35a228333a4c47ee.png"},{"id":80027538,"identity":"0b751eb4-d8e1-453b-af40-2ac97b1a5585","added_by":"auto","created_at":"2025-04-07 06:41:20","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":93908,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHeatmap of bacterial and fungal communities in the karst cave\u003c/strong\u003e.\u003cbr\u003e\n \u003cstrong\u003e(A)\u003c/strong\u003e Heatmap showing the distribution and abundance of bacterial species across the sampling points.\u003cbr\u003e\n \u003cstrong\u003e(B)\u003c/strong\u003e Heatmap illustrating the distribution and abundance of fungal species.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/0475941ada9a3b92520ac9fd.png"},{"id":80027723,"identity":"deecfdca-954d-42d6-9545-e96322e3dfdd","added_by":"auto","created_at":"2025-04-07 06:49:20","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":314574,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCircos plots of bacterial and fungal communities after treatment with plant-based removers in the karst cave\u003c/strong\u003e.\u003cbr\u003e\n \u003cstrong\u003e(A)\u003c/strong\u003e Circos plot showing the distribution and relationships of bacterial species following the application of three plant-based removers to lampenflora.\u003cbr\u003e\n \u003cstrong\u003e(B)\u003c/strong\u003e Circos plot illustrating the distribution and relationships of fungal species under the same treatment conditions. These plots highlight the impact of the plant-based removers on the microbial community structure, revealing changes in bacterial and fungal populations after the removal of lampenflora.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/dd12e80a4be5954c90a26649.png"},{"id":80026789,"identity":"6fe9dacf-d768-4dc3-baf5-32a988310b73","added_by":"auto","created_at":"2025-04-07 06:33:20","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":265503,"visible":true,"origin":"","legend":"\u003cp\u003eUnnumbered image in the Experimental Results section.\u003c/p\u003e","description":"","filename":"Unnumberfig.png","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/944c6b5234af630e7b9e4037.png"},{"id":84187893,"identity":"b2c5c614-7781-4fdb-bccd-c057ec68f278","added_by":"auto","created_at":"2025-06-09 06:03:12","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2162087,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6176730/v1/cc0f7e31-1323-40a1-b082-41c0f5aa2b28.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Study on the Inhibitory Effects of Plant Distillates on Microbial Communities in the lampenflora of Karst Tourist Caves","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCaves are typical representatives of karst landforms, usually formed by the gradual expansion of fissures in soluble rock masses by groundwater through dissolution and erosion processes, resulting in relatively closed and structurally stable underground spaces\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e. Since the temperature and humidity inside the cave are usually constant, these environments seem ideal. However, they are highly sensitive to external changes and human activities and exhibit low environmental carrying capacity, making them extremely vulnerable ecologically\u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/sup\u003e. Therefore, during cave development, such as building trails, installing lighting systems and other infrastructure, the originally closed natural structure will be disrupted. The resulting dust pollution and increased air exchange between the cave interior and exterior allow the invasion of exogenous organisms such as bacteria, archaea, fungi, lichens, insects, arachnids, and arthropods, posing a serious threat to the stability of the cave ecosystem. Furthermore, if cave management is inadequate after tourism development, issues such as the rapid proliferation of lampenflora under artificial lighting and seasonal increases in tourist numbers can lead to dramatic changes in environmental parameters like temperature, humidity, carbon dioxide concentration, and negative ion concentration. These changes may cause surface pollution, discoloration, and aging of the cave's landscape, further exacerbating ecological damage\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]\u003c/sup\u003e. These phenomena have become key environmental problems that need to be addressed in the development of cave tourism. Particularly in karst caves, the biological weathering caused by Lampenflora (referred to as \"lamp flora\") is especially prominent. These algae, mosses, and lichen communities typically grow within the range of artificial light sources, which provide them with suitable light and temperature conditions that promote their rapid reproduction. Depending on the intensity of light inside the cave, these lampenflora can be classified into different zones: the \u003cem\u003eChlorella\u003c/em\u003e and \u003cem\u003eChlorella-like\u003c/em\u003e algae zone, moss and lichen zone, lichen and algae zone, and fungi and algae zone \u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe growth and reproduction of lampenflora in karst caves primarily depend on organic matter residues within the cave, such as groundwater, animal excrement, and dust, hair brought in by tourists. These plants engage in complex ecological cycles through interactions between autotrophic and heterotrophic organisms\u003csup\u003e[8\u0026ndash;13,]\u003c/sup\u003e. At the same time, human activities, such as tourism, continually introduce additional plant and microbial species into the cave environment, making these community characteristics more prominent in tourist caves. The growth and metabolic activities of lampenflora significantly impact the surface layer of karst cave ecosystems. The organic acids secreted by these plants, along with organic matter left behind after their decay, form a porous layer on the rock surfaces within the cave, resulting in landscape corrosion and degradation of the cave's habitat for animals. Additionally, these metabolic products react with the calcium carbonate matrix, leading to the formation of precipitates that accelerate the enzyme-catalyzed dissolution of stalactites, with dissolution rates positively correlated to the activity of carbonic anhydrase (CA) \u003csup\u003e[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e. Furthermore, bacteria and fungi within the lampenflora communities have a particularly pronounced effect on the rock surfaces. Bacteria degrade the rock structure through the secretion of acidic substances, while fungal hyphae penetrate the mineral structure, contributing to the physical deterioration of stalactites. The metabolic by-products of both bacteria and fungi, especially organic acids, alter the local pH, further exacerbating rock degradation\u003csup\u003e[\u003cspan additionalcitationids=\"CR12 CR13\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]\u003c/sup\u003e. The colonization rates of microorganisms are closely related to the capillarity and roughness of the substrate. Smooth substrates retain moisture for longer periods but with lower moisture content, and while the initial colonization rate may be lower, microbial survival rates tend to be higher under reduced moisture conditions. As a result, lampenflora communities typically proliferate in moist, porous areas\u003csup\u003e[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]\u003c/sup\u003e. The spatial distribution of these communities is influenced by both light intensity and variations in temperature and humidity, which further affect community composition and metabolic activities, thus amplifying disturbances to the cave's microenvironment. Although the temperature and humidity within caves remain relatively constant, light intensity and distribution have a significant impact on the growth of lampenflora. When the light intensity decreases, the thickness of the biofilm will decrease accordingly. Therefore, addressing the issue of lampenflora is a critical component of cave ecosystem conservation\u003csup\u003e[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe control of lampenflora encompasses a variety of approaches, including physical, chemical, and emerging technologies. Physical methods, such as mechanical removal, utilize brushes or high-pressure water jets to remove the biofilms of lampenflora from rock surfaces. High-pressure water jets are particularly effective in areas with mature and structurally complex light-loving plant communities but may damage the mineral structure and promote the spread of algal spores, so they should be combined with other methods for optimal results \u003csup\u003e[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/sup\u003e. As a non-thermal effect technology, Radiofrequency (RF) technology uses electromagnetic waves to effectively reduce biofilm coverage without altering the rock matrix. It has achieved promising results in the laboratory and is expected to be further promoted in the future \u003csup\u003e[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/sup\u003e. Ultraviolet (UV-C) radiation, known for its strong bactericidal ability, is widely used in controlling lampenflora, while the new far UV-C technology, by controlling the wavelength, offers a balance between safety and efficiency, providing new possibilities for management \u003csup\u003e[\u003cspan additionalcitationids=\"CR22\" citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]\u003c/sup\u003e.Chemical methods primarily involve the use of quaternary ammonium salts, hydrogen peroxide, and chlorine-containing compounds to suppress or remove lampenflora. Benzalkonium chloride, while effective in bleaching, is difficult to degrade and may harm cave ecosystems \u003csup\u003e[\u003cspan additionalcitationids=\"CR25\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]\u003c/sup\u003e. Hydrogen peroxide is relatively eco-friendly and provides significant removal effects but may corrode calcite and lead to the recurrence of lampenflora \u003csup\u003e[\u003cspan additionalcitationids=\"CR28\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]\u003c/sup\u003e. Chlorine-based compounds, such as chlorine dioxide, have rapid decolorization capabilities but may have potential negative effects on cave-dwelling animals and minerals \u003csup\u003e[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]\u003c/sup\u003e. To address the limitations of traditional methods, combined physical and chemical approaches have become effective strategies in recent years. After using chemical agents to remove plants, high-pressure water jets are employed to clear any remaining residues, overcoming the drawbacks of single-method approaches.Moreover, plant extracts have emerged as an eco-friendly and sustainable control technology in recent years. Extracts from plants such as Artemisia annua, mugwort, and mint can adsorb nitrogen and phosphorus, reducing the nutrients required for biofilm growth. Active compounds in clove and cinnamon extracts can disrupt the cell membranes of lampenflora, effectively inhibiting the growth of algae and mosses \u003csup\u003e[\u003cspan additionalcitationids=\"CR30\" citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]\u003c/sup\u003e. Compared with traditional methods, plant extracts offer plant distillates have the advantages of high environmental adaptability and little impact on cave ecology, providing a green and sustainable solution for the management of karst cave lampenflora\u003csup\u003e[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn the context of biological weathering and environmental degradation caused by the excessive growth of lampenflora in karst caves, the scientific management of lampenflora has become a critical aspect of cave conservation. Plant extracts, as an emerging biological control method, offer significant advantages due to their excellent antibacterial properties, simple preparation process, low cost, and environmentally friendly characteristics. The natural compounds in these extracts can effectively inhibit the growth of lampenflora while avoiding secondary damage to cave ecosystems. According to previous studies, leaf extracts from cinnamon (Cinnamomum verum), mint (Mentha spp.), and mugwort (Artemisia argyi) have shown promising inhibitory effects \u003csup\u003e[\u003cspan additionalcitationids=\"CR35\" citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]\u003c/sup\u003e. Therefore, this study selected the distillation extracts of these three plants to systematically explore their potential applications potential in the removal of lampenflora, in order to provide a scientific evidence for the efficient management of lampenflora in karst caves and proposing feasible strategies for protecting cave ecosystems.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Study Area\u003c/h2\u003e \u003cp\u003eThe study was conducted in Zhijin Cave, located in Zhijin County, Guizhou Province, China. This area is a representative of karst landforms, with a long geological history and rich natural heritage. Due to the severe degradation caused by the growth of lampenflora, certain rocks in the cave have exhibited noticeable aesthetic deterioration and structural damage. The coverage of lampenflora not only affects the visual appeal of the cave but also accelerates the surface erosion of limestone, threatening the long-term conservation of the cave.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 In situ Experiment\u003c/h2\u003e \u003cp\u003eExperimental plots were established sequentially in the selected area, with each plot measuring 27\u0026times;27 cm and showing relatively complete coverage of lampenflora. Each plot was divided into a 3\u0026times;3 grid, where three different concentrations of plant distillation extracts were applied. We monitored the color changes of the plants at baseline (untreated), and then at 1 day, 7 days, and 30 days after treatment, using a portable precision colorimeter. The air temperature and humidity inside the cave were recorded using a hygrometer, while light intensity was measured with a light meter. The substrate pH value was tested using a PHS-25 pH meter. Since previous experiments showed that anhydrous ethanol had no significant effect on the color change of moss, no anhydrous ethanol control group was included in this study \u003csup\u003e[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Plant Sample Processing and Distillation Extract Preparation\u003c/h2\u003e \u003cp\u003eIn the experiment, the three plant samples purchased were first processed. The plant samples were washed with distilled water to remove surface dust and particles. After washing, the samples were air-dried naturally at room temperature (25\u0026deg;C), then wrapped in aluminum foil and dried in an oven at 60\u0026deg;C for 3 hours. Subsequently, the dried plant samples were ground into powder and sifted through a 40-mesh sieve (380 \u0026micro;m) to ensure sample uniformity. The plant powder was divided into three different amounts (5 g, 10 g, and 15 g), and each amount was mixed with 100 mL of anhydrous ethanol (99.7%) and heated to 78\u0026deg;C for distillation. The resulting clear plant distillation extracts were collected. Finally, the collected distillation extracts were stored in a refrigerator at 4\u0026deg;C to maintain their activity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Microbial Sampling\u003c/h2\u003e \u003cp\u003eThree representative sampling sites were selected in Zhijin Cave for microbial sampling. During the sampling process, sterile sampling bags and scrapers were used to collect surface microbial samples from different areas, including the cave walls, ceiling, and floor. After collection, all samples were immediately stored in a refrigerated box at 4\u0026deg;C and transported to the laboratory for DNA high-throughput sequencing analysis within 24 hours.\u003c/p\u003e \u003cp\u003eFor the experimental samples, the control points without treatment were labeled as Y points (including A1_1, A2_1, A3_1). The points treated with mint distillation extract were labeled as B points (including A1_3, A2_3, A3_3). The points treated with mugwort distillation extract were labeled as A points (including A1_4, A2_4, A3_4). The points treated with cinnamon distillation extract were labeled as R points (including A1_2, A2_2, A3_2).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Microbial Sequencing under Different Coverage Environments\u003c/h2\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.5.1 DNA Extraction\u003c/h2\u003e \u003cp\u003eMicrobial community genomic DNA was extracted according to the instructions provided by the E.Z.N.A.\u0026reg; Soil DNA Kit (Omega Bio-tek, Norcross, GA, U.S.). The quality of the extracted genomic DNA was assessed using 1% agarose gel electrophoresis. DNA concentration and purity were determined using the FastDNA\u0026reg; Spin Kit for Soil (MP Biomedicals, U.S.).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e2.5.2 PCR Amplification and Sequencing Library Construction\u003c/h2\u003e \u003cp\u003eThe bacterial 16S rRNA gene region (A1_1-A3_3) was amplified using the primers 338F (5\u0026rsquo;-ACTCCTACGGGAGGCAGCAG-3\u0026rsquo;) and 806R (5\u0026rsquo;-GGACTACHVGGGTWTCTAAT-3\u0026rsquo;). The PCR reaction system included: 4 \u0026micro;L of 5\u0026times; FastPfu buffer, 2 \u0026micro;L of 2.5 mM dNTPs, 0.8 \u0026micro;L of upstream primer (5 \u0026micro;M), 0.8 \u0026micro;L of downstream primer (5 \u0026micro;M), 0.4 \u0026micro;L of FastPfu DNA polymerase, 0.2 \u0026micro;L of BSA, 10 ng of template DNA, and the final volume adjusted to 20 \u0026micro;L. The fungal ITS rRNA gene ITS1 region was amplified using primers ITS1F (5\u0026rsquo;-CTTGGTCATTTAGAGGAAGTAA-3\u0026rsquo;) and ITS2R (5\u0026rsquo;-GCTGCGTTCTTCATCGATGC-3\u0026rsquo;). The PCR reaction system included: 2 \u0026micro;L of 10\u0026times; rTaq buffer, 2 \u0026micro;L of 2.5 mM dNTPs, 0.8 \u0026micro;L of upstream primer (5 \u0026micro;M), 0.8 \u0026micro;L of downstream primer (5 \u0026micro;M), 0.2 \u0026micro;L of rTaq DNA polymerase, 0.2 \u0026micro;L of BSA, 10 ng of template DNA, and the final volume adjusted to 20 \u0026micro;L. The amplification program was as follows: denaturation at 95\u0026deg;C for 3 minutes, followed by 27 cycles (35 cycles for fungi) of 95\u0026deg;C for 30 seconds, 55\u0026deg;C for 30 seconds, and 72\u0026deg;C for 45 seconds, with a final extension at 72\u0026deg;C for 10 minutes, and stored at 4\u0026deg;C (PCR machine: ABI GeneAmp\u0026reg; 9700).The purified PCR products were then used for library construction with the NEXTFLEX Rapid DNA-Seq Kit: (1) adapter ligation, (2) magnetic bead selection to remove adapter-dimer fragments, (3) PCR amplification for library enrichment, (4) magnetic bead recovery of the PCR product to obtain the final library. Sequencing was performed on the Illumina Nextseq2000 platform (Shanghai Meiji Biotechnology Co., Ltd.).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.5.3 Sequencing Data Processing\u003c/h2\u003e \u003cp\u003eThe paired-end raw sequencing reads were quality-controlled using fastp software, and merged using FLASH software:(1) Base calls with quality scores below 20 at the tail of the reads were filtered. A 50 bp sliding window was used, and if the average quality score in the window was below 20, the bases at the end of the window were trimmed. Reads shorter than 50 bp and reads containing N bases were also removed after quality control.(2) Paired-end reads were merged based on their overlap relationship, with a minimum overlap length of 10 bp.(3) The maximum mismatch ratio allowed in the overlap region of the merged sequence was set to 0.2, and sequences that did not meet this threshold were filtered out.(4) The barcode and primer at both ends of the sequence were used to distinguish samples and adjust the sequence direction. No mismatches were allowed in the barcode, while a maximum of 2 mismatches were allowed in the primer region.To minimize the impact of sequencing depth on subsequent Alpha and Beta diversity analyses, the number of sequences in all samples was rarefied, and diversity, community composition, and other analyses were performed based on the rarefied OTU (Operational Taxonomic Unit) data.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e2.5.4 DATA AVAILABILITY STATEMENT\u003c/h2\u003e \u003cp\u003eThe raw data have been submitted to NCBI Sequence Read Archive (SRA) under Bioproject accession PRJNA1222770.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"3. Experimental Results","content":"\u003cp\u003e\u003cstrong\u003e3.1 Microbial Community \u0026alpha;-Diversity Analysis and the Inhibitory Effects of Plant Distillates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMicrobial \u0026alpha;-diversity was chosen to evaluate the differences in microbial communities across the three different coverage environments. The Chao index and Shannon index were used to represent the bacterial diversity, respectively.\u003c/p\u003e\n\u003cp\u003eBy analyzing the Chao and Shannon indices of microorganisms in the biofilms of cave\u0026nbsp;lampenflora, the effects of different removal agents on the richness and diversity of microbial communities can be clearly observed.\u003c/p\u003e\n\u003cp\u003eBased on both the Chao and Shannon indices, bacterial richness and diversity underwent significant changes after applying different removers. As shown in the figure, the Y point exhibited higher richness and diversity. However, after using the removers, particularly B and A, both the Chao and Shannon indices showed a marked decrease, indicating the most significant reduction in bacterial richness at point A. In comparison, the treatment effect at point R was milder, suggesting that certain bacterial phyla might exhibit tolerance to cinnamon distillate, even showing adaptive reproduction in the removal environment.\u003c/p\u003e\n\u003cp\u003eIn contrast to bacteria, the Chao and Shannon indices of fungi were significantly lower, indicating that the richness and diversity of fungi were relatively limited. After applying the removers, particularly at point A, the Chao and Shannon indices of fungi significantly decreased, showing that the wormwood distillate had the strongest inhibitory effect on fungi. In contrast, the inhibitory effects of mint and cinnamon distillates were relatively weaker, with mint distillate potentially exhibiting selective inhibition of certain fungal genera.\u003c/p\u003e\n\u003cp\u003eThese results suggest that the three plant distillates demonstrated varying efficacy in removing\u0026nbsp;lampenflora\u0026nbsp;and their associated microorganisms. Among them, wormwood distillate exhibited the most significant antibacterial and antifungal effects, making it the most suitable candidate for microbial management in caves.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.2 Disruption of Microbial Community Structure in Karst Caves by Plant Distillates and Heatmap Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHeatmap analysis was conducted to investigate the impact of plant distillates on the bacterial and fungal communities in Karst caves, revealing the patterns and characteristics of microbial community changes under different treatment conditions.\u003c/p\u003e\n\u003cp\u003eThe heatmap of the bacterial community showed that the untreated group (Y) formed an independent cluster, indicating that the bacterial community structure remained unaffected by external interventions. In contrast, the groups treated with plant distillates (R, B, A) clustered together, demonstrating a significant impact of the reagents on the bacterial community structure. Specifically, the use of cinnamon distillate (R) reduced the abundance of genera such as \u003cem\u003enorank_f__Spirosomaceae\u003c/em\u003e, \u003cem\u003eCellvibrio\u003c/em\u003e, and \u003cem\u003ePaenibacillus\u003c/em\u003e, while increasing the abundance of \u003cem\u003ePedomicrobium\u003c/em\u003e and \u003cem\u003eLeptolyngbya_ANT.L52.2\u003c/em\u003e. The use of mugwort distillate (A) significantly decreased the abundance of \u003cem\u003eScytonema_PCC-7110\u003c/em\u003e and \u003cem\u003enorank_f__A4b\u003c/em\u003e, while increasing the abundance of genera such as \u003cem\u003eAcinetobacter\u003c/em\u003e and \u003cem\u003ePseudomonas\u003c/em\u003e. The mint distillate (B) reduced the abundance of \u003cem\u003eHyphomicrobium\u003c/em\u003e and \u003cem\u003eScytonema_PCC-7110\u003c/em\u003e, while significantly increasing the abundance of \u003cem\u003ePseudomonas\u003c/em\u003e and \u003cem\u003eMassilia\u003c/em\u003e. These changes indicate that plant distillates can effectively disrupt the composition and metabolic function of bacterial communities in environments covered by light-sensitive plants.\u003c/p\u003e\n\u003cp\u003eThe heatmap of the fungal community showed a more pronounced trend of changes. The mugwort distillate (A) treatment group formed a distinct cluster, indicating a significant impact on the fungal community, while the untreated group (Y) clustered together with the cinnamon (R) and mint (B) treatment groups, reflecting similar changes in fungal communities induced by these treatments. Specifically, the mugwort distillate (A) significantly reduced the abundance of genera such as \u003cem\u003eEntoloma\u003c/em\u003e, \u003cem\u003eTrichoderma\u003c/em\u003e, and \u003cem\u003ePenicillium\u003c/em\u003e, while increasing the abundance of genera like \u003cem\u003eFurcasterigmium\u003c/em\u003e, \u003cem\u003eMortierella\u003c/em\u003e, and \u003cem\u003ePlectosphaerella\u003c/em\u003e. The mint distillate (B) decreased the abundance of \u003cem\u003eLeptobacillium\u003c/em\u003e and \u003cem\u003eAscotricha\u003c/em\u003e, but significantly increased the abundance of \u003cem\u003eVerticillium\u003c/em\u003e and \u003cem\u003eBrunneochlamydosporium\u003c/em\u003e. The cinnamon distillate (R) reduced the abundance of \u003cem\u003eRozellomycota_gen_Incertae_sedis\u003c/em\u003e and \u003cem\u003eTrichoderma\u003c/em\u003e, while increasing the abundance of \u003cem\u003eMetapochonia\u003c/em\u003e and \u003cem\u003eSimplicillium\u003c/em\u003e. Overall, the fungal community exhibited a more sensitive response to the plant distillates, resulting in significant adjustments in community structure.\u003c/p\u003e\n\u003cp\u003eOverall, the surface microenvironment of the karst cave\u0026rsquo;s lampenflora and the biofilm-covered areas formed a short-term microbial community succession process. The use of plant distillates altered the abundance and structure of microorganisms, with bacteria showing a more significant response to cinnamon and mint distillates, while the fungal community underwent a marked adjustment under mugwort distillate treatment. The results suggest that plant distillates can effectively interfere with the microbial community dynamics in light plant-covered environments, particularly exhibiting the most significant inhibitory effect on fungi.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.3 Regulatory Effect of Plant Distillates on the Microbial and Fungal Community Structure in Karst Caves\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBefore the plant distillate treatments (as shown in Figures A and B), the microbial community in the karst cave was dominated by bacteria. The phylum \u003cem\u003eProteobacteria\u003c/em\u003e (47.0%) was the absolute dominant group, while \u003cem\u003eCyanobacteria\u003c/em\u003e (10.7%) and \u003cem\u003eChloroflexi\u003c/em\u003e (9.2%) also exhibited relatively high abundance. The presence of cyanobacteria indicates that, under the light plant-covered environment, photosynthetic microorganisms occupy a certain proportion of the cave ecosystem. However, these microbial groups may accelerate rock weathering through photosynthesis, leading to carbonate dissolution and negatively impacting the structural integrity of cave walls\u003csup\u003e\u0026nbsp;[36]\u003c/sup\u003e. In the fungal community, \u003cem\u003eAscomycota\u003c/em\u003e (60.9%) was the dominant phylum, followed by \u003cem\u003eBasidiomycota\u003c/em\u003e (15.3%) and \u003cem\u003eRozellomycota\u003c/em\u003e (14.6%).\u003c/p\u003e\n\u003cp\u003eAfter the plant distillate treatments, Figure A shows that the bacterial community composition underwent significant changes. Following cinnamon distillate (R) treatment, the abundance of \u003cem\u003eProteobacteria\u003c/em\u003e increased to 63.3%, while the abundance of \u003cem\u003eCyanobacteria\u003c/em\u003e significantly decreased to below the detection limit, indicating that cinnamon distillate effectively inhibited the photosynthetic microbial group. Additionally, the abundance of \u003cem\u003eActinobacteriota\u003c/em\u003e increased to 11.0%, suggesting an enrichment of microorganisms involved in organic matter decomposition.In contrast, both peppermint distillate (B) and mugwort distillate (A) treatments led to bacterial communities with higher diversity. Notably, after mugwort distillate treatment, the abundance of \u003cem\u003eActinobacteriota\u003c/em\u003e and \u003cem\u003eFirmicutes\u003c/em\u003e reached 13.2% and 6.5%, respectively, indicating a significant promotion of microorganisms responsible for the breakdown of complex organic matter.\u003c/p\u003e\n\u003cp\u003eFigure B illustrates the changes in fungal communities after treatment with plant distillates. After cinnamon distillate treatment, the abundance of \u003cem\u003eAscomycota\u003c/em\u003e significantly increased to 91.4%, while the abundance of \u003cem\u003eRozellomycota\u003c/em\u003e decreased to 0.4%, indicating a strong inhibitory effect of cinnamon distillate on parasitic fungi.After treatment with peppermint distillate and mugwort distillate, the abundance of \u003cem\u003eBasidiomycota\u003c/em\u003e increased to 8.6% and 31.8%, respectively. Mugwort distillate also significantly increased the abundance of \u003cem\u003eMortierellomycota\u003c/em\u003e to 12.9%, demonstrating its promoting effect on the microbial group responsible for decomposing organic matter.\u003c/p\u003e\n\u003cp\u003eFigures A and B visually demonstrate the significant regulatory effects of plant distillates on the structure of microbial and fungal communities. Cinnamon distillate exhibits a strong inhibitory effect, significantly reducing the abundance of harmful microbial groups such as \u003cem\u003eCyanobacteria\u003c/em\u003e and \u003cem\u003eRozellomycota\u003c/em\u003e. In contrast, peppermint and mugwort distillates show better community diversity maintenance, while also promoting the enrichment of functional microbial groups such as \u003cem\u003eActinobacteriota\u003c/em\u003e and \u003cem\u003eBasidiomycota\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4. Spectral Characteristics Analysis of Plant Distillates and Their Mechanisms for Inhibiting Lampenflora in Karst Tourist Caves\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBy analyzing the relationship between absorbance peaks and concentrations of mugwort, peppermint, and cinnamon distillates using ultraviolet spectrophotometry, we can not only gain a more detailed understanding of the active components responsible for removing lampenflora in karst tourist caves but also infer their activity variations and underlying mechanisms at different concentrations.\u003c/p\u003e\n\u003cp\u003eThe absorbance peak at 208 nm is primarily attributed to organic acids (such as benzoic acid), phenolic compounds, and flavonoids present in mugwort. As the concentration of mugwort increases, the concentration of these active components also rises, thereby enhancing the absorption of ultraviolet light and increasing mugwort\u0026apos;s activity in plant removal. This phenomenon suggests that the natural chemical constituents of mugwort exhibit stronger UV absorption at higher concentrations, potentially providing effective support for its plant inhibitory effects.\u003c/p\u003e\n\u003cp\u003eThe figure below shows the absorption spectra of peppermint distillate at different concentrations within the wavelength range of 200 nm to 400 nm.\u003c/p\u003e\n\u003cp\u003eThe absorbance peaks of peppermint primarily occur at wavelengths of 207 nm and 229 nm. As the concentration increases, the absorbance at these two wavelengths gradually increases, demonstrating a positive correlation between concentration and absorbance peak values.At these wavelengths, the main active components in peppermint, such as menthol and peppermint oil, exhibit significant UV absorbance characteristics.Menthol and peppermint oil, as volatile organic compounds, possess strong UV absorption capacity, particularly at the 207 nm and 229 nm wavelengths.With increasing concentration, the concentration of these active components rises, further enhancing peppermint\u0026apos;s UV light absorption ability at these two wavelengths.Furthermore, the active components in peppermint not only effectively absorb UV light but may also exert inhibitory effects on plants by releasing volatile chemicals. This effect becomes more pronounced as the concentration of peppermint increases.\u003c/p\u003e\n\u003cp\u003eThe figure below shows the absorption spectra of cinnamon distillate at different concentrations within the wavelength range of 200 nm to 400 nm.\u003c/p\u003e\n\u003cp\u003eThe absorbance peaks of cinnamon occur at 207 nm and 286 nm, with a more pronounced change in absorbance observed at 286 nm.As the concentration increases, the absorbance peak values at both wavelengths show a significant increase, with the increase in absorbance being positively correlated with concentration.This indicates that the active components in cinnamon, such as cinnamic acid, cinnamaldehyde, and coumarin, exhibit more pronounced UV absorption characteristics as the concentration increases.These components in cinnamon not only have strong antioxidant, antimicrobial, and plant growth-regulating effects, but as UV absorption increases, they may enhance the release of active components, improve biological activity, and produce stronger inhibitory effects during plant removal.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThis study explores the potential of plant distillates in managing lampenflora in karst caves, with a particular focus on their inhibitory effects on microbial communities associated with lampenflora.The results show that the distillates of peppermint, mugwort, and cinnamon effectively inhibit the growth of lampenflora in karst caves to varying extents and significantly impact the diversity and structure of microbial communities.\u003c/p\u003e\n\u003cp\u003eFirst, plant distillates have a significant effect on the α-diversity of microbial communities.α-diversity analysis revealed that mugwort distillate exhibited the most significant decrease in bacterial and fungal diversity, particularly in the Chao and Shannon indices.Mugwort distillate exhibited the strongest inhibitory effects on microbial communities, demonstrating potent antibacterial and antifungal activity.\u003c/p\u003e\n\u003cp\u003eFurther spectral analysis revealed strong absorbance signals in the 290-310 nm range for mugwort distillate. This phenomenon may be closely related to the presence of flavonoids, phenolic compounds, and certain organic acids in mugwort.Flavonoids and phenolic compounds are widely distributed in plants and are known for their significant antimicrobial, antioxidant, and UV shielding properties.Under UV light exposure, these components can effectively absorb UV light, reducing the risk of harmful UV radiation to cells. At the same time, through interactions with the cell membrane, they interfere with normal cellular functions, thereby affecting microbial growth and metabolism.Flavonoids inhibit microbial growth by suppressing photosynthesis, disrupting the integrity of cell membranes, or interfering with key metabolic pathways.Therefore, these chemical components in mugwort distillate may be the main reason for its strong inhibitory effect on microbial community diversity.\u003c/p\u003e\n\u003cp\u003eThe inhibitory effects of peppermint and cinnamon distillates are relatively mild. Peppermint distillate may exert inhibitory effects through its volatile components, showing a certain influence on microbial communities, especially at higher concentrations.The inhibitory effect of cinnamon distillate is more complex. Its spectral characteristics show a prominent absorbance peak in the 250-290 nm range, especially within the 280-300 nm range.The absorbance characteristics in this range may be related to the presence of coumarin compounds in cinnamon, which have been shown to possess some antimicrobial properties.Further analysis of the inhibitory mechanism of cinnamon distillate may help to better understand its impact on microbial communities.\u003c/p\u003e\n\u003cp\u003eMugwort distillate, due to its rich content of flavonoids and phenolic compounds, exhibits a strong antimicrobial effect.Peppermint and cinnamon distillates exert inhibitory effects through other components. Although their effects are not as pronounced as that of mugwort, they still impact the microbial community.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThe results show that all three distillates (mint, Artemisia, and cinnamon) inhibited cave plant growth at certain concentrations, with Artemisia distillate having the strongest effect on both plants and microbes. It reduced plant growth and significantly decreased microbial diversity, particularly in bacteria and fungi, highlighting its antimicrobial properties. Mint and cinnamon also inhibited growth, with mint selectively affecting certain fungi.\u003c/p\u003e\n\u003cp\u003eThrough the analysis of experimental data, it is evident that plant distillate, as a green and environmentally friendly inhibitor, possess considerable application potential. They can not only effectively inhibit the excessive growth of light plants, but also reduce the richness and diversity of microbial communities, thereby slowing down the biological weathering process in karst caves. It provides a sustainable solution for the protection and restoration of cave environment.\u003c/p\u003e\n\u003cp\u003eIn summary, the experimental results of this study validate the effectiveness of plant distillates in the management of light plants in karst caves. Moreover, this method demonstrates good environmental adaptability and can reduce negative impacts on the cave ecosystem, making it a promising approach for further promotion and application. Future research could explore the combined effects of different plant distillates under various environmental conditions, optimize management strategies, and assess their long-term effects and sustainability.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eZhiyi Xu: Investigation, Software, Writing - Original Draft.Mingzhong Long: Project Administration, Funding Acquisition, Conceptualization, Methodology, Supervision, Writing - Original Draft, Writing - Review \u0026amp; Editing.Shengyu Yang: Data Curation, Methodology.Jinru Liu: Formal Analysis, Resources.Kehua Wu: Funding Acquisition, Investigation, Methodology.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eThis work was supported by the Basic Research Program of Guizhou Province(Qiankehe Jichu ZK [2023] Yiban 147) ; the Research Fund of Guizhou Minzu University (GZMUZK[2024]QD64).\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003ePlease reference PRJNA1222770 in your publication. This BioProject accession number is provided instead of SRP and should be used in your publication as it will allow better searching in Entrez.Accession to cite for these SRA data: PRJNA1222770Temporary Submission ID: SUB15085079Release date: 2025-02-12\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eKatja Sterflinger, Guadalupe Pi\u0026ntilde;ar.Microbial deterioration of cultural heritage and worksof art \u0026mdash; tilting at windmills?Appl Microbiol Biotechnol,2013,97:9637\u0026ndash;9646\u003c/li\u003e\n \u003cli\u003eNata\u0026scaron;a Nikolić, Gordana Subakov Simić, Igor Golić,Slađana Popović.The effects of biocides on the growth of aerophytic green algae (Chlorella sp.) isolated from a cave environment[J].Arch Biol Sci. 2021;73(3):341-351\u003c/li\u003e\n \u003cli\u003eZhu Xiaoyan, Zhang Meiliang. Study on Cave Environmental Factors in Karst Cave Tourism Activities [J]. China Karst, 2020, 39(03): 426-431.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eValme Jurado , Yolanda del Rosal , Jose Luis Gonzalez-Pimentel ,et al.Biological Control of Phototrophic Biofilms in a ShowCave: The Case of Nerja Cave[J].Applied sciences,2020, 10, 3448; doi:10.3390/app10103448\u003c/li\u003e\n \u003cli\u003eT.Aley. Tourist Caves: Algae and Lampenflora[C]//John.Gunn, Encyclopaedia of Caves and Karst Science. New York:Fitzroy Dearborn，2004:733-734.\u003c/li\u003e\n \u003cli\u003eEsteban R P .Study and remediation of environmental problems caused due to the growth of algae in speleothems of calcareous caves adapted for tourism- a case of success in Spain [J].Journal of Environmental Geology,2018,02(01): \u0026nbsp;\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eNata\u0026scaron;a Nikolić,C, Nikola Zarubica , Bojan Gavrilović,et al.LAMPENFLORA AND THE ENTRANCE BIOFILM IN TWO SHOW CAVES:COMPARISON OF MICROBIAL COMMUNITY,ENVIRONMENTAL,AND BIOFILM PARAMETERS.[J]Journal of Cave and Karst Studies, v. 82, no. 2, p. 69-81. DOI:10.4311/2018EX0124\u003c/li\u003e\n \u003cli\u003eChen Weihai, Deng Yadong, Tang Li, et al. Study on the Sustainable Use of Tourism Caves in Guilin [J]. Guangxi Science, 2018, 25(05): 579-589. DOI: 10.13656/j.cnki.gxkx.20181106.003Slađana Popović, Marija Pećić ,Gordana Subakov Simić.Exploring Lampenflora of ResavskaCave, Serbia.Biology and life sciences forum.2022, 15, 33. https://doi.org/10.3390/IECD2022-12425\u003c/li\u003e\n \u003cli\u003eSlađana Popović, Marija Pećić ,Gordana Subakov Simić.Exploring Lampenflora of ResavskaCave, Serbia.Biology and life sciences forum[J].2022, 15, 33. https://doi.org/10.3390/IECD2022-12425\u003c/li\u003e\n \u003cli\u003eZhang Kaiyan, Li Tongjian, Zhang Xianqiang, et al. The Carbonic Anhydrase of Three Lithophytic Moss Species and Its Erosion Effect on Limestone [J]. China Karst, 2017, 36(04): 441-446.\u003c/li\u003e\n \u003cli\u003eJake,Robin ,Jacob J , et al.Lampenflora in a Show Cave in the Great Basin Is Distinct from Communities on Naturally Lit Rock Surfaces in Nearby Wild Caves[J].Microorganisms,2021,\u003c/li\u003e\n \u003cli\u003eEsteban R P .Study and remediation of environmental problems caused due to the growth of algae in speleothems of calcareous caves adapted for tourism- a case of success in Spain [J].Journal of Environmental Geology,2018,02(01):\u003c/li\u003e\n \u003cli\u003eHern\u0026aacute;ndez-Marin\u0026eacute; M. \u0026amp; Rold\u0026aacute;n M., Adherence of hormogonia to substrata is mediated by polysaccharides produced by necridic cells. Algological Studies,2005,117:239-249.\u003c/li\u003e\n \u003cli\u003eMulec J ,Kosi G ,Vrhovsek D .ALGAE PROMOTE GROWTH OF STALAGMITES AND STALACTITES IN KARST CAVES (SKOCJANSKE JAME, SLOVENIA)[J].Carbonates And Evaporites,2007,22(1):6-9.\u003c/li\u003e\n \u003cli\u003eMiller A.Z., Dion\u0026iacute;sio A., Laiz L., Macedo M.F. \u0026amp; Saiz-Jim\u0026eacute;nez C., The influence of inherent properties of building limestones on their bioreceptivity to phototrophic microorganisms. Annals of Microbiology,59: 705-713. https://doi.org/10.1007/BF03179212\u003c/li\u003e\n \u003cli\u003eDavid G . Caves:Processes, Development and Management[M]. Blackwell Publishing Ltd.: 1996-11-17. DOI:10.1002/9781444313680.\u003c/li\u003e\n \u003cli\u003eHern\u0026aacute;ndez-Marin\u0026eacute;\u0026nbsp;M ,Rold\u0026aacute;n M .Adherence of hormogonia to substrata is mediated by polysaccharides produced by necridic cells[J].Algological Studies/Archiv f\u0026uuml;r Hydrobiologie, Supplement Volumes,2005,117239-249.\u003c/li\u003e\n \u003cli\u003eP C ,P C ,A G , et al. Biofilms on tuff stones at historical sites: identification and removal by nonthermal effects of radiofrequencies. [J]. Microbial ecology, 2013, 66 (3): 659-68.\u003c/li\u003e\n \u003cli\u003eSt\u0026eacute;phane P ,Olympe E ,Battle K , et al.UV-C as an efficient means to combat biofilm formation in show caves: evidence from the La Glaci\u0026egrave;re Cave (France) and laboratory experiments.[J].Environmental science and pollution research international,2017,24(31):24611-24623.\u003c/li\u003e\n \u003cli\u003eThomas L. Kieft, Devyn Del Curto, Zo\u0026euml; Havlena,et al.Potential for Mitigation of Cave Lampenflora Using BenzalkoniumChloride or UV-C.Geoheritage (2023) 15:68https://doi.org/10.1007/s12371-023-00839-4\u003c/li\u003e\n \u003cli\u003eM. Buonanno, B. Ponnaiya, D. Welch, M. Stanislauskas, G. Randers-Pehrson,L. Smilenov, F.D. Lowy, D.M. Owens, D.J. Brenner, Germicidal efficacy andmammalian skin safety of 222-nm UV light, Radiat. Res. 187 (4) (2017) 493\u0026ndash;501,https://doi.org/10.1667/rr0010cc.1.\u003c/li\u003e\n \u003cli\u003eEsteban R P . Study and remediation of environmental problems caused due to the growth of algae in speleothems of calcareous caves adapted for tourism- a case of success in Spain [J]. Journal of Environmental Geology, 2018, 02 (01): Spain\u003c/li\u003e\n \u003cli\u003eRosangela A ,Daniela B ,Beatriz C , et al.A multidisciplinary approach to the comparison of three contrasting treatments on both lampenflora community and underlying rock surface.[J].Biofouling,2023,39(2):11-14.\u003c/li\u003e\n \u003cli\u003eSt\u0026eacute;phane P ,Thomas M ,Faisl B , et al.Bleaching of biofilm-forming algae induced by UV-C treatment: a preliminary study on chlorophyll degradation and its optimization for an application on cultural heritage.[J].Environmental science and pollution research international,2018,25(14):14097-14105.\u003c/li\u003e\n \u003cli\u003eEsteban R P . Study and remediation of environmental problems caused due to the growth of algae in othems of calcareous caves adapted for tourism- a case of success in Spain [J]. Journal of Environmental ogy, 2018, 02 (01): Spain\u003c/li\u003e\n \u003cli\u003eFaimon J ,\u0026Scaron;telcl J ,Kube\u0026scaron;ov\u0026aacute;\u0026nbsp;S , et al.Environmentally acceptable effect of hydrogen peroxide on cave\u0026nbsp;\u0026ldquo;lamp-flora\u0026rdquo;, calcite speleothems and limestones[J].Environmental Pollution,2003,122(3):417-422.\u003c/li\u003e\n \u003cli\u003eNatasa N ,Subakov G S ,Igor G , et al.The effects of biocides on the growth of aerophytic green algae (Chlorella sp.) isolated from a cave environment[J].ARCHIVES OF BIOLOGICAL SCIENCES,2021,73(3):341-351.\u003c/li\u003e\n \u003cli\u003eHe Huiyan. Study on the Inhibition of Algal Cell Growth, Metabolic Activity, and Photosynthesis by Hydrogen Peroxide [D]. Xi\u0026apos;an University of Architecture and Technology, 2020. DOI: 10.27393/d.cnki.gxazu.2020.001237.\u003c/li\u003e\n \u003cli\u003eDuc Anh Trinh, Quan Hong Trinh, Ngoc Tran,et al.Eco-friendly Remediation of Lamp enflora on Sp Eleothems in Tropical Karst Caves.Journal of Cave and Karst Studies, v. 80, no. 1, p. 1-12 DOI: 10.4311/2017ES0101\u003c/li\u003e\n \u003cli\u003eAddesso R ,Bellino A ,D\u0026apos;Angeli M I , et al.Vermiculations from karst caves: The case of Pertosa-Auletta system (Italy)[J].Catena,2019,182104178-104178.\u003c/li\u003e\n \u003cli\u003eMeng Hui, Xu Yong. GC-MS Analysis of Volatile Oil Components in Fresh Artemisia argyi Leaves from Shanghai [J]. Journal of Pharmaceutical Practice, 2009, 05 - 0362 - 03.\u003c/li\u003e\n \u003cli\u003eWu Fan, Jia Ruhan. Research Progress on the Pharmacological Effects of Cinnamon Extract [J]. Herald of Medicine, 2012, 31(7): 882-885.\u003c/li\u003e\n \u003cli\u003eZheng Yao, Yang Xiaoxi, Qian Xinyu, Zhu Yuejie, Chen Jiachang, Xu Pao. Study on the Water Purification and Antibacterial Effects of Chinese Herbal Medicines. Environmental Science and Technology, 43(1), 222-228. DOI: 10.19672/j.cnki.1003-6504.2020.01.033.\u003c/li\u003e\n \u003cli\u003eLin Jiayu, Li Bangjiang, Cheng Cai, et al. Current Research on the Biological Weathering Prevention and Control of Stone Buildings [J]. Journal of Applied Ecology, 2021, 32(08): 3023-3030. DOI: 10.13287/j.1001-9332.202108.039\u003c/li\u003e\n \u003cli\u003eYao Yongfang, Shi Lin, Tan Caideng. Study on the Extraction of Antibacterial Substances from Artemisia argyi [J]. Food Science and Technology, 2011, 36(11). DOI: 10.13684/j.cnki.spkj.2011.11.004\u003c/li\u003e\n \u003cli\u003eYang Cuiyun, An Xin, Wan Jingqiong, et al. Food Science and Technology [J], 2021, 46(1). DOI: 10.13684/j.cnki.spkj.2021.01.030\u003c/li\u003e\n \u003cli\u003eLi Ping, Shi Chuntao, Shu Ting, Shen Xiaoxia. Comparison of Antibacterial Activity of Cinnamon Oil Extracted by Three Different Methods [J]. Preservation and Processing, 2018, 18(2): 31-38. DOI: 10.3969/j.issn.1009-6221.2018.02.006\u003c/li\u003e\n \u003cli\u003eFaimon J. Environmentally acceptable effect of hydrogen peroxide on cave \u0026quot;lampenflora\u0026quot;. Calcite speleothems and limestones. Environmental Pollution, Elsevier Science Ltd. 2003:417-22.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Karst Caves, lampenflora, Ecological Restoration, Cave Ecosystem","lastPublishedDoi":"10.21203/rs.3.rs-6176730/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6176730/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCurrently, the ecosystems of karst caves faces serious prpblems of biological weathering and environmental damage caused by lampenflora. This issue is exacerbated by artificial lighting sources and tourist activities in cave tourism development, which promote the excessive growth of these plants and result in the degradation of cave rocks and the surrounding ecological environment. Therefore, the management of lampenflora is a key task in cave ecosystem conservation. This study explores the inhibitory effects of plant distillates, specifically from mint, mugwort, and cinnamon, on lampenflora in karst caves as an eco-friendly and sustainable solution. Through experiments in Zhijin Cave, Guizhou Province, the inhibitory effects of different plant distillates at different concentrations on lampenflora communities and their microbial communities were studied. \u0026nbsp;The results showed that mugwort distillate had the most significant inhibitory effect on bacterial and fungal communities, with microbial diversity and richness significantly reduced in samples treated with mugwort distillate. Mint and cinnamon distillates showed relatively weaker effects, with mint exhibiting selective inhibition on certain fungal species. Overall, plant distillates, particularly mugwort distillate, demonstrated great potential in protecting cave ecosystems, inhibiting lampenflora, and controlling associated microbial communities. The findings confirm that plant distillates can effectively suppress the growth of lampenflora, reduce microbial community richness and diversity, and thereby slow down the biological weathering process in caves. This research provides a new and sustainable solution for the ecological management in karst caves, which has high application value and environmental adaptability.\u003c/p\u003e","manuscriptTitle":"Study on the Inhibitory Effects of Plant Distillates on Microbial Communities in the lampenflora of Karst Tourist Caves","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-07 06:33:15","doi":"10.21203/rs.3.rs-6176730/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8aba8a10-b819-4277-8d63-755192578ca3","owner":[],"postedDate":"April 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":46739841,"name":"Biological sciences/Ecology/Microbial ecology"},{"id":46739842,"name":"Earth and environmental sciences/Environmental social sciences/Sustainability"}],"tags":[],"updatedAt":"2025-06-09T05:39:04+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-07 06:33:15","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6176730","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6176730","identity":"rs-6176730","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}