Mold Aspergillus Research Articles

Synthesis, characterization and antimicrobial activity of nalidixic acid derivatives with spirohydantoins

This article presents the synthesis of a series of amides, based on the interaction of several 3-aminospirohydantoins with nalidixic acid. The target compounds were characterized by physicochemical parameters, IR, 1H and 13C NMR spectral data. The antimicrobial activity of the products obtained was determined against Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis, Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa and Salmonella abony, the yeasts Candida albicans and Saccharomyces cerevisiae and the molds Penicillium chrysogenum and Aspergillus niger. The relationship between structure and biological activity of the products obtained was discussed. It was found that the most effective compounds are tetralin (5f) and indane (5g) derivatives, which exhibit a pronounced antimicrobial activity against both tested Gram-positive and Gram-negative bacteria.

Chemical and Bacterial Components in Sake and Sake Production Process.

Together with the worldwide Washoku (traditional Japanese foods and drinks) boom, interest in sake, a traditional Japanese alcoholic drink, is increasing around the world. There are few scientific analyses and studies on the production of sake or the final product itself. We show the diversity of bacterial contaminants during sake production and investigated the effects of different ingredients on sake (for example, amino acids). The koji mold Aspergillus oryzae converts rice starch into sugars, and then, the sake yeast Saccharomyces cerevisiae converts the sugars to ethanol. Comparative studies of the bacterial flora of different sakes have shown that various bacterial species are detected, but that there are few frequently detected bacteria. In addition, the bacterial flora does not vary much during the process of sake brewing, after the koji (steamed rice covered with koji mold) and moto (fermentation starter) are mixed, suggesting that most bacteria contaminate the sake during the process of koji and moto production. Thus, there is the possibility that the contaminating bacteria may grow due to a relationship with the koji mold and/or the sake yeast. The flavor, taste, and quality of sakes differ, even between the same brands of sakes, which may be attributed to variations in the contaminating bacteria during sake production.

COP9 Signalosome Interaction with UspA/Usp15 Deubiquitinase Controls VeA-Mediated Fungal Multicellular Development.

COP9 signalosome (CSN) and Den1/A deneddylases physically interact and promote multicellular development in fungi. CSN recognizes Skp1/cullin-1/Fbx E3 cullin-RING ligases (CRLs) without substrate and removes their posttranslational Nedd8 modification from the cullin scaffold. This results in CRL complex disassembly and allows Skp1 adaptor/Fbx receptor exchange for altered substrate specificity. We characterized the novel ubiquitin-specific protease UspA of the mold Aspergillus nidulans, which corresponds to CSN-associated human Usp15 and interacts with six CSN subunits. UspA reduces amounts of ubiquitinated proteins during fungal development, and the uspA gene expression is repressed by an intact CSN. UspA is localized in proximity to nuclei and recruits proteins related to nuclear transport and transcriptional processing, suggesting functions in nuclear entry control. UspA accelerates the formation of asexual conidiospores, sexual development, and supports the repression of secondary metabolite clusters as the derivative of benzaldehyde (dba) genes. UspA reduces protein levels of the fungal NF-kappa B-like velvet domain protein VeA, which coordinates differentiation and secondary metabolism. VeA stability depends on the Fbx23 receptor, which is required for light controlled development. Our data suggest that the interplay between CSN deneddylase, UspA deubiquitinase, and SCF-Fbx23 ensures accurate levels of VeA to support fungal development and an appropriate secondary metabolism.

Diagnostic-driven management of invasive fungal disease in hematology in the era of prophylaxis and resistance emergence: Dutch courage?

Patients receiving intensive anti-leukemic treatment or recipients of allogeneic hematopoietic stem cell transplantation (HSCT) are prone to develop invasive fungal disease caused by both Aspergillus and non-Aspergillus moulds. Overall mortality following invasive mould disease (IMD) is high; adequate and timely antifungal treatment seems to ameliorate the outcome, yet early diagnosis in the haematological patient remains a challenge for most clinicians. Prophylaxis and the empiric addition of antifungal therapy to neutropaenic patients with fever persisting or recurring during broad-spectrum antibiotic treatment is therefore standard of care in many institutions. However, aside from the potential for overtreatment and important side effects, the emergence of resistance to medical triazoles in Aspergillus fumigatus poses a risk for inadequate initial treatment. Initial voriconazole therapy in patients with azole-resistant invasive aspergillosis was recently shown to be associated with a 23% increased mortality rate compared to the patients with azole-susceptible infection, despite changing to appropriate antifungal therapy once resistance was detected. Moreover, fever is not always present with IMD; therefore, cases may be missed when relying solely on this symptom for starting diagnostic procedures and antifungal treatment. At our institution, a diagnostic-driven treatment approach for IMD was implemented relying on clinical but also laboratory markers to start antifungal treatment. We describe the basis and clinical implementation of our diagnostic-driven approach in this review.

The effect of molase and urea on the production of alkaline protease from indigenous Aspergillus flavus DUCC K225

Protease is one of industrial important enzyme which can be produced by microorganisms. including fungi. Alkaline protease had physiological and commercially important. this enzyme used in detergent industries as additives. The indigenous mold Aspergillus flavus DUCC K225 isolated from lime soil of Madura island demonstrate the ability in producing extracellular protease at pH 8. The aim of this study was to examined the production of alkaline protease from A. flavus K225 on Czapeks Dox modified liquid medium containing molase as C-source and urea as N-source. The study were done in submerged culture on rotary shaker. The production of alkaline protease were observed after 7 days incubation. by examined fungal biomass. protease activity. protein content and protease specific activity. The examination of enzyme activity conducted at pH 8.5 done on crude enzyme and ammonium phosphate fractions of 0-20%. 20-40%. 40-60%. 60-80% and 80-100%. The results showed that the fungal biomass gained was 14.27 g/L. while the highest protease specific activity found at 40-60% ammonium sulphate fractination (71.75 U/ml) and 4.2 purification fold.

Modulation of Fatty Acid Composition of Aspergillus oryzae in Response to Ethanol Stress.

The koji mold Aspergillus oryzae is widely adopted for producing rice wine, wherein koji mold saccharifies rice starch and sake yeast ferments glucose to ethanol. During rice wine brewing, the accumulating ethanol becomes a major source of stress for A. oryzae, and there is a decline in hydrolysis efficiency. However, the protective mechanisms of A. oryzae against ethanol stress are poorly understood. In the present study, we demonstrate that ethanol adversity caused a significant inhibition of mycelium growth and conidia formation in A. oryzae, and this suppressive effect increased with ethanol concentration. Transmission electron microscopy analysis revealed that ethanol uptake triggered internal cellular perturbations, such as irregular nuclei and the aggregation of scattered vacuoles in A. oryzae cells. Metabolic analysis uncovered an increase in fatty acid unsaturation under high ethanol conditions, in which a large proportion of stearic acid was converted into linoleic acid, and the expression of related fatty acid desaturases was activated. Our results therefore improve the understanding of ethanol adaptation mechanisms in A. oryzae and offer target genes for ethanol tolerance enhancement via genetic engineering.

Cellulose products modified with monomeric and gemini surfactants: antimicrobial aspects

The aim of this study was to examine the impact of microbiocides on the reduction of the microbial population on paper in order to protect it against biodeterioration. The cellulose products were modified with the cationic gemini surfactant hexamethylene-1,6-bis-(N,N-dimethyl-N-dodecylammonium bromide) C6 and its monomeric analog (didecyldimethylammonium chloride) DDAC. The microbiocides were introduced into the paper by coating and spraing. In the coating method the microbiocides were mixed with starch solution and applied to paper surface as a wet film with a thickness of 24 and 50 µm. In the spraying method the surfactants were applied at 3% concentration in water to get 0.005 ml/cm2 of paper. Experiments were performed with moulds of the genera Aspergillus, Penicilllium, Chaetomium and Trichoderma and bacteria represented by the genera Bacilllus and Pseudomonas. The antimicrobial properties of the paper modified with surfactants were determined by qualitative and quantitative methods. The zones of inhibition were measured using the parallel streak method. Macroscopic assessment of mould growth on the surface of the paper samples was performed over 21 days of incubation. In quantitative analysis, the survival rate of microorganisms on the modified paper samples was determined over 24 h. Aspergillus brasiliensis was the least sensitive mould, with no observable inhibition zones. No growth of this mould was observed either on or underneath the sample. Both of the surfactants applied as coatings or sprays protected the paper effectively against the growth of bacteria and mould. However, the spraying method is simpler to use. Over 24 h, the number of spores and bacteria in all the samples containing surfactants was reduced to below 1 log/10 cm2 (reduction factor up to 99.9%). The compounds may therefore be applied as antimicrobial agents for the protection of paper.Graphical abstract

Association of molds and metrological parameters to frequency of severe asthma exacerbation

BackgroundSensitization to airborne molds may be a risk factor for severe asthma and direct cause of asthma exacerbation (AE).MethodsA prospective, 1-year (April 2016–March 2017) study, done in Kuwait Allergy Centre, investigated the link between AEs with exposure to outdoor molds and the role of meteorological parameters in mold sensitized patients and compared with non-allergic asthma patients who had asthma deterioration. The total of 676 adult asthmatics with moderate-severe AEs were included and divided into atopic (85.65%) and non-atopic group. Atopy was defined by positive skin prick test (SPT) to at least one inhalant allergen. Data regarding atopy and asthma severity were collected from patient’s records. Patients with symptoms and signs of acute respiratory infection and patient sensitized to indoor allergens only were excluded. Daily count of local pollens (Salsola kali, Bermuda grass) and molds (Aspergillus, Alternaria and Cladosporium) were obtained from the Aerobiology department. Daily metrological parameters (atmospheric pressure-AP, temperature-T and relative humidity-RH) were provided by Kuwait Environment Public Authority. Count of spores/m3 and weather variable are shown on weekly basis. The year circle was divided into 4 Seasons (1, 2, 3, 4) accordingly to typical desert climate.ResultsSensitization to molds was relatively high but significantly less (25.0%) if compared to the pollens sensitization. The highest number of AEs was in season 4 for both molds and pollens sensitized patients. Seasonal patterns for both allergens were significant and positively correlated with RH and AP. In season 1 only, mold sensitized patients showed higher rate of AEs. Non-atopic patients have been less sensitive to increased RH than atopic. Negative correlation with T was similar in both atopic and non-atopic patients.ConclusionDespite of high rate of sensitization to molds, their significant role in triggering AE was not found in desert environment. Typical desert climate and high allergencity of local weeds outweigh the influence of the molds.

Effect of pepA deletion and overexpression in Aspergillus luchuensis on sweet potato shochu brewing

The mash of sweet potato shochu (Japanese distilled spirit) has a low pH value because the shochu koji mold produces a large amount of citric acid, which prevents germ contamination. In this study, we examined acid protease PepA's role in shochu production. For this purpose, we constructed pepA deletion and overexpression strains, using a black koji mold Aspergillus luchuensis RIB 2604 (NBRC 4314), with the Agrobacterium-mediated transformation method. The rice koji, prepared using a pepA disruptant (ΔpepA) and pepA-overexpressing strain (OEpepA), demonstrated 1/2- and 24-fold acid protease activities compared to that prepared using the parental strain, respectively. A small-scale test of sweet potato shochu brewing indicated the mash of ΔpepA had a lower amino acid concentration, while the mash of OEpepA had a higher concentration than that produced by the parental strain. Therefore, the mash amino acid concentrations were proportional to these strains' acid proteases activities. After distilling these mashes, we examined each shochu's aroma components. Shochu prepared using ΔpepA had relatively higher aroma components, such as alcohol and ester, compared to that prepared using parental strains. Meanwhile, shochu prepared using OEpepA had lower aroma components than that prepared using the parental strains. Based on these results, the amount of shochu aroma components showed an inverse correlation to the acid protease activity in the mash. Thus, the koji mold's acid protease content had a greater influence on the aroma qualities of sweet potato shochu. Accordingly, we have discussed the possibility of the breeding of shochu koji mold with acid protease as an indicator.

The Antimicrobial and Wound Healing Potential of Opuntia ficus indica L. inermis Extracted Oil from Tunisia.

Introduction Opuntia ficus indica L. inermis (OFI) is used in traditional medicine pharmacopeia for its richness in natural bioactive compounds. It has been proven to be effective in the improvement of the healing of laser-induced skin burns. The aim of the present study was to investigate the wound healing effect of OFI extracted oil on full-thickness skin wound. Materials and Methods The OFI seeds were firstly isolated from mature prickly pears, washed, dried, and then cold-pressed. The antimicrobial activities of OFI seed oil were estimated in vitro against bacteria, yeast, and fungi. Minimum Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were calculated. Skin wound healing was investigated using an excisional wound healing model in rats. The skin wounds of three experimental groups of rats were topically treated once/day with saline solution (control group), 0.15 mg/mm2 of a reference drug Esth'Elle Pharma Cicaplaie cream (reference group), and 0.6 μl/mm2 of OFI seed oil (OFI oil group). The healing process was monitored daily and the percentage of wound contraction was calculated. A histological study was carried on skin biopsies. Results The extracted oil has shown an interesting antimicrobial effect on Enterobacter cloacae, antiyeast effect against Candida parapsilosis and Candida sake, and antifungal activity against three opportunistic cutaneous molds (Penicillium, Aspergillus, and Fusarium). Moreover, OFI oil has shown a good wound healing effect. It prevents cutaneous infections and reduces the reepithelialization phase. Conclusion OFI extracted oil has in vitro antimicrobial/fungal properties and in vivo wound healing activity. It seems to be efficient in the treatment of cutaneous infections and the promoting of the scarring process.

Intracellular Delivery of Biologically-Active Fungal Metabolite Gliotoxin Using Magnetic Nanoparticles.

Gliotoxin (GT), a secondary metabolite produced by Aspergillus molds, has been proposed as a potential anti-tumor agent. Here we have developed a nanoparticle approach to enhance delivery of GT in tumor cells and establish a basis for its potential use as therapeutical drug. GT bound to magnetic nanoparticles (MNPs) retained a high anti-tumor activity, correlating with efficient intracellular delivery, which was increased in the presence of glucose. Our results show that the attachment of GT to MNPs by covalent bonding enhances intracellular GT delivery without affecting its biological activity. This finding represents the first step to use this potent anti-tumor agent in the treatment of cancer.

Labeled quantitative mass spectrometry to study the host response during aspergillosis in the common bottlenose dolphin (Tursiops truncatus)

Aspergillosis is a fungal infection caused by Aspergillus molds that can affect both humans and animals. Despite advances in diagnostics and therapy, medical management of this disease remains difficult. Expansion of the basic knowledge regarding its pathophysiology in animals is critical to aid in the identification of new biomarkers of infection for diagnosis and therapeutic targets. For such a purpose, proteomics can be used by addressing protein changes during various disease processes. In the present study, a mass spectrometry analysis based on isobaric tagging for relative and absolute quantitation (iTRAQ®) was applied for direct identification and relative quantitation of proteins in blood collected from 32 Aspergillus-diseased common bottlenose dolphins (Tursiops truncatus, 32 samples) in comparison with blood from 55 other dolphins (55 samples from 41 clinically-normal controls and from 14 cetaceans with miscellaneous non-Aspergillus inflammation diseases) and ten convalescent dolphins (28 samples). Sixty-six and 40 proteins were found to be ≥2.0-fold over- and underrepresented versus miscellaneous non-Aspergillus inflammatory dolphins, respectively, and most were confirmed vs. clinically-normal controls and convalescents. Many proteins which play a role in the adaptive immune response were identified, including MHC proteins and others involved in catalytic activity like the NADPH-ubiquinone oxido-reductases. Overall, iTRAQ® appears to be a convenient proteomic tool greatly suited for exploratory ex vivo studies focusing on pathophysiology. This technique should be considered as a preliminary step before validation of new diagnostic markers.

Antifungal Activity of Some Ethnomedicinally Important Tuberous Plants of Family Liliaceae

Fungal spores are often present in air and soil which may cause internal as well as external infections. Phytochemicals are extracted from plants of family Liliaceae and identified having antifungal and antibiotic properties. The present study deals antifungal activity of crude extract of leaf of five species Asparagus L. and four species Chlorophytum Ker. Gawl. of family Liliaceae using a different polar and non-polar solvent like methanol, petroleum ether and acetone. Effects of different plant extract were tested on yeast (Candida albicans) and mold (Aspergillus niger) using potato dextrose agar medium by agar well diffusion method. Zone of inhibition produced by different plant extracts was calculated. Among these plants, Chlorophytum borivilianum Santapau and Fernandes., Chlorophytum tuberosum Baker. and Asparagus racemosus Willd. show maximum antifungal activities against two clinical fungi Candida albicans and Aspergillus niger. In case of three solvent, acetone plant extract showed a significant reduction, while methanol showed its minimum reduction in the growth of these two opportunist fungi. Streptomycin was used as control drug for antifungal studies.

Learning to control tissue damage while fighting Aspergillus.

Aspergillus moulds are increasingly being recognised as significant human pathogens that can cause life-threatening infections in the context of host immune dysregulation, particularly in the lung. It is now clear that there is a close relationship between infection susceptibility and the fine regulation of pulmonary immunity and inflammation. While the contribution of IL-17/Th17 responses to both physiological and pathological lung inflammation is now well established, the cellular interactions, soluble factors, and signalling pathways that determine Th17 cell responses to fungal infection remain unclear. Here, we identify potential key mediators of fungus-DC-T cell interactions in the respiratory tract, with a focus on the DC-derived cytokines thought to exert a major influence on generation of pathological Th17 cells. We review recent data indicating a crucial role for Aspergillus-induced autophagy in lung DCs on subsequent T-cell polarization and modulation of 'stemness', which appears critical for avoiding pathological lung inflammation and promoting disease resolution.

Antimicrobial spectrum activity of bacteriocinogenic Staphylococcus strains isolated from goat and sheep milk

Bacteriocins have attracted great attention as potential alternatives to antibiotics and chemical food additives. In the present study, 243 Staphylococcus isolates from milk samples (n = 110) of goat and sheep herds located in Fars province, Iran, were screened for antimicrobial substance production. Twenty-eight isolates showed an antagonistic activity against the indicator strain Micrococcus luteus ATCC 4698. The susceptibility of all antimicrobial substances to proteolytic enzymes allowed us to consider them as bacteriocin-like inhibitory substances (BLIS). The term BLIS is applied to uncharacterized proteinaceous antimicrobials produced by gram-positive bacteria. Based on molecular identification methods, the isolates belonged to the species Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus pseudintermedius, Staphylococcus aureus, and Staphylococcus agnetis. Pulsed-field gel electrophoresis revealed a high level of genotype diversity among the Staph. chromogenes isolates. All of the isolates harbored nukA or bsaA2 genes, suggesting that their BLIS were related to nukacin or Bsa. The antimicrobial compounds from test strains were not effective against gram-negative pathogens, including Salmonella enterica serovar Typhimurium, Escherichia coli O157:H7, and Klebsiella pneumonia as well as the indicator mold Aspergillus fumigatus. All the gram-positive targets, including Bacillus cereus, Listeria monocytogenes, Enterococcus faecalis Ef37 (a tyramine-producer strain), Lactobacillus saerimneri 30a (a histamine-producer strain), and methicillin-resistant Staph. epidermidis, were inhibited by the Staph. chromogenes isolates. Staphylococcus haemolyticus 4S12 was able to inhibit the majority of gram-positive bacteria. Listeria monocytogenes strains were the only indicators sensitive to the antimicrobial agents produced by Staph. agnetis 4S97B. The other Staphylococcus strains were ineffective on all the organisms tested. Based on their inhibitory capacities, the BLIS produced by the Staph. chromogenes isolates seem to be interesting candidates for developing novel antimicrobial agents.

Impairing fluoride export of Aspergillus fumigatus mitigates its voriconazole resistance

Fungi have evolved specific export activities to balance intracellular levels of the toxic ion fluoride, while the first-line antimycotic voriconazole contains fluorine. This study aimed to explore whether impaired fluoride export might result in altered susceptibilities of the human pathogenic mould Aspergillus fumigatus towards this antifungal compound. Functional characterization of the putative fluoride exporter in A. fumigatus was performed in the context of azole resistance by generating deletion strains that were assessed for their resistance against fluoride and voriconazole. The FexA fluoride exporter of A. fumigatus appears to be expressed constitutively, and targeting its encoding gene results in significantly increased sensitivity towards this halide. Impaired fluoride export correlates with increased susceptibility of an azole-resistant fexAΔ strain. These results demonstrate that the fexA-encoded gene product is the major fluoride export activity of A. fumigatus, and that voriconazole serves as a source of fluoride. However, these data do not support the application of voriconazole based on fluoride toxicity.

Resistance of concern in fungi

Antifungal drugs save lives by treating serious fungal infections, just as antibacterial drugs (antibiotics) are used to treat bacterial infections. This is especially a concern for patients with invasive infections like those caused by the fungus Candida, a yeast, or the mold Aspergillus, which can cause serious health problems, including death. There are indications that reports of antifungal resistance are increasing among common fungal species. Some species of fungi are naturally resistant to treatment with certain types of antifungal medications. Other species can develop resistance over time due to improper antifungal use—for example, dosages too low or treatment courses that are not long enough. Some studies have indicated that antibacterial medications may also contribute to antifungal resistance. This resistance could occur for a variety of reasons. For example, antibacterial drugs can reduce good and bad bacteria in the gut, which creates favourable conditions for Candida growth. It is not yet known if decreasing the use of all or certain antibiotics can reduce Candida infections, but appropriate use of antibacterial and antifungal medications is one of the most important factors in fighting drug resistance. More information is needed about the risk antifungal resistance poses on human health and how many people are sickened by drug-resistant fungal infections each year. CDC and its partners are working to better understand why and how antifungal resistance emerges and to increase awareness among medical and public health communities about these infections. Ultimately, we need to develop better methods to prevent and control drug-resistant fungal infections.

Design of biosystems to provide healthy and safe food—part B: effect on microbial flora and sensory quality of orange juice

This study was performed to determine the antifungal activity of plant oil extracts (1% of lemongrass and oregano, 1:1) in combination with organic citrus fruit extract (BIOSECUR F440D®) with a ratio of 1:3 (W/W) against pathogenic molds (Aspergillus niger, Penicillium chrysogenum) and yeast (Saccharomyces cerevisiae). Synthetic commercial preservatives such as sodium benzoate (E211) and potassium sorbate (E202) were used as references. Antifungal formulation-loaded microemulsion was prepared by microfluidization for in situ analysis in orange juice. Antifungal formulation-loaded microemulsion (AFF-MM) led to a total inhibition of targeted fungi during the first days of storage and their populations stayed below detection limit until the end of storage (35 days). AFF-MM presented a higher antifungal activity compared to synthetic preservatives even at a concentration of 10 times lower than optimized concentration. Indeed, microemulsion increased the bioactivity and bioavailability of plant oil/citrus extracts compared to coarse emulsion by increasing the surface area of droplets induced by size decrement. Finally, the sensory analysis showed the panelists acceptance of orange juice in presence of antifungal-loaded microemulsion.

Identification of biofilm composition covering lime-based materials

This paper aims at identification of biofilms composition taken from lime and lime-cement plasters. The samples of plasters were exposed to natural weathering for more than three years to weather conditions of Prague as a representative of Central European continental climate. After biofilms sampling, they were cultivated using specific nutrient solutions and analysed using various types of microscopes. It was found, that the plasters were covered mostly by green algae (Hematococcus pluvialis, Klebsormidium sp., Trebouxiophyceae, Chlorophyceae and Chlorellaceae), cyanobacteria (Nostoc, Anabaena, Leptolyngbya, Phormidium, Plectonema, Synechococcales and Oscillatoriale), moulds (Aspergillus niger, Alternaria) and moss (Ceratodon purpureus). Possible influence of these organisms on hygrothermal behaviour of the base materials is subsequently discussed.

Effect of high temperature with Litsea cubeba Pers. to control mold growth on bamboo food packaging and its possible modes of action

This study examined the effect of high temperature and Litsea cubeba oil with its main components against molds (Aspergillus niger, Aspergillus flavus, Penicillium sp., Penicillium cyclopium, Rhizopus sp., Fusarium sp., and Cladosporium sp.) on bamboo food packaging. Response surface methodology (RSM) with X1 (concentration of L. cubeba oil at 100, 300, and 500 mg g-1), X2 (temperature at 60, 80, and 100 °C), and X3 (time at 12, 14, and 16 h) was used to find the inhibitory periods of natural mold on packaging plates. The physical properties and the change of chemical components on the bamboo packaging plate before and after temperature treatment were determined to find the mode of action using gas chromatography–mass spectrometry (GC-MS). High temperature (at 100 °C) was a good inhibitor of all mold growth with the MIC (minimum inhibitory concentration) of 300 mg g-1; without heat treatment, no MIC was found. In addition, it was found that by using 300 mg g-1 of L. cubeba oil at 100 °C with an exposure time of 12 h, spore germination on the bamboo surfaces was completely inhibited for at least 290 days. After using high temperature, citral was detected on the surface of the packaging plates. Therefore, these components could be the key factors for inhibiting molds.