Solar/ClO2 system inactivates fungal spores in drinking water: Synergy, efficiency and mechanisms.

The risk of fungal pollution in drinking water has been paid attention. Solar/chlorine dioxide (ClO2) combined system is an environment-friendly, economical and efficient disinfection method, especially for countries and regions that are economically backward and still exposed to unsafe drinking water. In this paper, the kinetics, influencing factors, mechanism and regrowth potential of inactivated Aspergillus niger (A. niger) spores by solar/ClO2 were reported for the first time. The inactivation curve can be divided into three stages: instant inactivation within 1-2 min, slow linear inactivation and finally a tail. The synergistic factors produced by solar/ClO2 in terms of log reduction and maximum inactivation rate were 1.194 and 1.112, respectively. The inhibitory effect on the regrowth of A. niger spores inactivated by solar/ClO2 was also stronger than that by ClO2 alone. Strongly oxidizing reactive species produced by solar/ClO2 accelerated the accumulation of endogenic reactive oxygen species (ROS) caused by oxidation stress of A. niger spores, improving the inactivation ability of the system. The inactivation order of A. niger spores was: loss of culturability, accumulation of intracellular ROS, loss of membrane integrity, leakage of intracellular species and change of morphology. The inactivation performance of solar/ClO2 was better than solar/chlor(am)ine according to the comparison of inactivation efficiency and regrowth potential. Results also suggested that solar/ClO2 process was more suitable for the treatment of ground water sources.

Current and Future Effects of Climate Change on Airborne Allergens.

PURPOSE OF REVIEW: Delineation of the impact of elevated carbon dioxide and concomitant global warming on airborne allergens is performed. RECENT FINDINGS: European tree pollen trends in general showed earlier start and end dates and increased total pollen release, with some differences both in locale and among species. Earlier flowering was also seen with grasses and weeds. In the case of some boreal trees, flowering was delayed due to a pre-seasonal requirement for necessary accumulated chilling temperature to achieve bud-set. Anthropogenic climate change induced rise in temperature and CO2 levels has resulted in demonstrable increases in aeroallergens. This has been most dramatic in tree pollen annual load, but also seen with grasses and weeds. Collected data is greatest for the Northern Hemisphere, especially the European continent, with supporting data from North America and Australia.

Fungi: friends or foes-an outreach science initiative for the collection of airborne fungal spores by high school students.

Fungi mostly reproduce through spores that are adapted for airborne dispersal; hence, fungal spores (and fungi) are found virtually everywhere. Fungi can be "friends or foes." Our friends include fungi used in the food and biotech industries, fungi that contribute to the cycling of carbon and nutrients, and those involved in the decontamination of polluted soils and/or water, to mention just a few examples. Many species, however, are foes-they are detrimental to plants, animals, and/or humans. Annually, >1.5 million people die due to invasive fungal infections. With the aim of enhancing microbiology literacy and the understanding of microbial concepts, we set up a project for the collection of airborne spores (the principal agent through which human airways are exposed to fungi). Students from five high schools in the Oeiras municipality partnered with us as citizen scientists; they carried out sampling by collecting fungal spores on adhesive stickers. The fungal spores collected by the students were subsequently processed in the schools and our research laboratory. Results obtained by the students themselves revealed a large variety of fungal species capable of growing in a rich medium at 30°C. In the research laboratory, using selective isolation conditions, 40 thermotolerant fungi were isolated, 32 of which were taxonomically identified as aspergilla, mostly from within the Aspergillus fumigatus taxa, yet exhibiting high genetic heterogeneity. The protocols and results were presented to the students, who were made aware of the local dispersal of airborne fungal spores, including some from potentially pathogenic fungi. Through carrying out scientific activities, the students developed both the interest and the self-confidence needed to implement future environmental investigations.

Automatic real-time monitoring of fungal spores: the case of Alternaria spp.

We present the first implementation of the monitoring of airborne fungal spores in real-time using digital holography. To obtain observations of Alternaria spp. spores representative of their airborne stage, we collected events measured in the air during crop harvesting in a contaminated potato field, using a Swisens Poleno device. The classification algorithm used by MeteoSwiss for operational pollen monitoring was extended by training the system using this additional dataset. The quality of the retrieved concentrations is evaluated by comparison with parallel measurements made with a manual Hirst-type trap. Correlations between the two measurements are high, especially over the main dispersion period of Alternaria spp., demonstrating the potential for automatic real-time monitoring of fungal spores.

The aggregation characteristics of Aspergillus spores under various conditions and the impact on LPUV inactivation: Comparisons with chlorine-based disinfection.

Aggregation is the primary step prior to fungal biofilm development. Understanding the attributes of aggregation is of great significance to better control the emergence of waterborne fungi. In this study, the aggregation of Aspergills spores (A. flavus and A. fumigatus) under various salt, culture medium, and humic acid (HA) conditions was investigated for the first time, and the inactivation via low-pressure ultraviolet (LPUV) upon aggregated Aspergillus spores was also presented. The aggregation efficiency and size of aggregates increased over time and at low salt (NaCl and CaCl2) concentration (10 mM) while decreasing with the continuous increase of salt concentration (100 and 200 mM). Increasing the concentration of culture medium and HA promoted the aggregation of fungal spores. Spores became hydrated, swelled, and secreted more viscous substances during the growth period, which accelerated the aggregation process. Results also suggested that fungal spores aggregated more easily in actual water, posing a high risk of biohazard in real-life scenarios. Inactivation efficiency by LPUV decreased with higher aggregation degrees due to the protection from the damaged spores on the outer layer and the shielding of pigments in the cell wall. Compared to chlorine-based disinfection, the aggregation resulted in the extension of shoulder length yet neglectable change of inactivation rate constant under LPUV treatment. Further investigation of cell membrane integrity and intracellular reactive oxygen species was conducted to elucidate the difference in mechanisms between various techniques. This study provides insight into the understanding and controlling of the aggregation of fungal spores.

Compatible solutes determine the heat resistance of conidia.

BACKGROUND: Asexually developed fungal spores (conidia) are key for the massive proliferation and dispersal of filamentous fungi. Germination of conidia and subsequent formation of a mycelium network give rise to many societal problems related to human and animal fungal diseases, post-harvest food spoilage, loss of harvest caused by plant-pathogenic fungi and moulding of buildings. Conidia are highly stress resistant compared to the vegetative mycelium and therefore even more difficult to tackle. RESULTS: In this study, complementary approaches are used to show that accumulation of mannitol and trehalose as the main compatible solutes during spore maturation is a key factor for heat resistance of conidia. Compatible solute concentrations increase during conidia maturation, correlating with increased heat resistance of mature conidia. This maturation only occurs when conidia are attached to the conidiophore. Moreover, conidia of a mutant Aspergillus niger strain, constructed by deleting genes involved in mannitol and trehalose synthesis and consequently containing low concentrations of these compatible solutes, exhibit a sixteen orders of magnitude more sensitive heat shock phenotype compared to wild-type conidia. Cultivation at elevated temperature results in adaptation of conidia with increased heat resistance. Transcriptomic and proteomic analyses revealed two putative heat shock proteins to be upregulated under these conditions. However, conidia of knock-out strains lacking these putative heat shock proteins did not show a reduced heat resistance. CONCLUSIONS: Heat stress resistance of fungal conidia is mainly determined by the compatible solute composition established during conidia maturation. To prevent heat resistant fungal spore contaminants, food processing protocols should consider environmental conditions stimulating compatible solute accumulation and potentially use compatible solute biosynthesis as a novel food preservation target.

Annual Change in Fungal Concentrations and Allergic Sensitization Rates to Alternaria and Cladosporium in Korea During the Period 1998-2022.

PURPOSE: Atmospheric fungi are associated with respiratory allergies in humans, and some fungal spores can cause allergic diseases. Environmental and biological factors influence the concentrations of atmospheric spores. In this study, we evaluated the climate change-induced annual variations in fungal spore concentrations and allergic sensitization rates in the Seoul Metropolitan Area over a period of 25 years. METHODS: Fungal spores and pollen were obtained from Hanyang University Seoul and Guri Hospitals; they were identified and counted for 25 years (1998-2022). The study participants included patients who underwent tests for allergic diseases in both hospitals. Their allergenic sensitization rates were determined via allergic skin prick and serum tests, after which their sensitization rates to allergenic fungi and pollens were calculated. The daily climatic variables were obtained from the Korea Meteorological Administration. RESULTS: The total annual atmospheric fungal concentrations decreased in both areas during the period. Simultaneously, we recruited 21,394 patients with allergies (asthma, 1,550; allergic rhinitis, 5,983; and atopic dermatitis, 5,422) from Seoul and Guri Hospitals for allergenic fungal sensitization evaluations over the period. The allergenic fungal sensitization rates decreased annually in both areas over that time `+(Alternaria [3.5%] and Cladosporium [4.4%] in 1998; Alternaria [0.2%] and Cladosporium [0.2%] in 2022). In contrast, the annual pollen concentrations increased with the sensitization rates to pollen in children. CONCLUSIONS: The atmospheric fungal concentrations decreased annually, with allergic sensitization rate decreasing over the period of 25 years. Allergenic fungal sporulation could decrease with climate changes, such as desertification and drought. Extended monitoring periods and further large-scale studies are required to confirm the causality and to evaluate the impact of climate change.

Discovery, Identification, and Insecticidal Activity of an Aspergillus flavus Strain Isolated from a Saline-Alkali Soil Sample.

Aphids are one of the most destructive pests in agricultural production. In addition, aphids are able to easily develop resistance to chemical insecticides due to their rapid reproduction and short generation periods. To explore an effective and environmentally friendly aphid control strategy, we isolated and examined a fungus with aphid-parasitizing activity. The strain (YJNfs21.11) was identified as Aspergillus flavus by ITS, 28S, and BenA gene sequence analysis. Scanning electron microscopy and transmission electron microscopy revealed that the infection hyphae of 'YJNfs21.11' colonized and penetrated the aphid epidermal layer and subsequently colonized the body cavity. Field experiments showed that 'YJNfs21.11' and its fermentation products exerted considerable control on aphids, with a corrected efficacy of 96.87%. The lipase, protease, and chitinase secreted by fungi help aphid cuticle degradation, thus assisting spores in completing the infection process. Additionally, changes were observed in the mobility and physical signs of aphids, with death occurring within 60 h of infection. Our results demonstrate that A. flavus 'YJNfs21.11' exhibits considerable control on Aphis gossypii Glover and Hyalopterus arundimis Fabricius, making it a suitable biological control agent.

Fungal spores in Caribbean mangrove sediments, dataset from southeastern Mexico.

Most paleoecological investigations use different biotic or abiotic proxies for climate and environmental reconstructions. Although fossil pollen is one of the most used biological proxies, Non-Pollen Palynomorphs (NPPs), especially fungal spores and tissues, have an underestimated potential to infer local and regional climate dynamics. This dataset describes the most common Non-pollen palynomorphs of fungal origin from mangrove sediments in the Caribbean Sea, southeastern Mexico. A detailed descriptive Atlas is presented, with light micrographs taken from routine pollen slides in paleoecological reconstructions. Microphotographs were included to facilitate their identification. A total of 59 spores, 4 tissues, 2 hyphae, and 11 unidentified fungal palynomorphs are described.

Outdoor airborne allergens: Characterization, behavior and monitoring in Europe.

Aeroallergens or inhalant allergens, are proteins dispersed through the air and have the potential to induce allergic conditions such as rhinitis, conjunctivitis, and asthma. Outdoor aeroallergens are found predominantly in pollen grains and fungal spores, which are allergen carriers. Aeroallergens from pollen and fungi have seasonal emission patterns that correlate with plant pollination and fungal sporulation and are strongly associated with atmospheric weather conditions. They are released when allergen carriers come in contact with the respiratory system, e.g. the nasal mucosa. In addition, due to the rupture of allergen carriers, airborne allergen molecules may be released directly into the air in the form of micronic and submicronic particles (cytoplasmic debris, cell wall fragments, droplets etc.) or adhered onto other airborne particulate matter. Therefore, aeroallergen detection strategies must consider, in addition to the allergen carriers, the allergen molecules themselves. This review article aims to present the current knowledge on inhalant allergens in the outdoor environment, their structure, localization, and factors affecting their production, transformation, release or degradation. In addition, methods for collecting and quantifying aeroallergens are listed and thoroughly discussed. Finally, the knowledge gaps, challenges and implications associated with aeroallergen analysis are described.

Solar-driven strongly coupled plasmonic Au nanoarrays on mesoporous silica nanodisks enable selective fungal and bacterial inactivation in well water.

Well water is an important water source in isolated rural areas but easily suffers from microbial contamination. Herein, we anchored periodic Au nanoarrays on mesoporous silica nanodisks (Au-MSN) to fabricate a solar-driven nano-stove for well water disinfection. The solar/Au-MSN process completely inactivated 3.98, 6.55, 7.11 log10 cfu/mL, and 3.37 log10 pfu/mL of Aspergillus niger spores, Escherichia coli, chlorine-resistant Spingopyxis sp. BM1-1, and bacteriophage MS2 within 5 min, respectively. Moreover, the complete inactivation of various microorganisms (even at a viable but nonculturable state) was achieved in the flow-through reactor under natural solar light in real well water matrixes. Thorough characterizations and theoretical simulations verified that the densely anchoring strategy of Au-MSN's nanoarray worked on broadband absorption via the photon confinement effect, and trace amounts of Au can induce strong electromagnetic fields and collective localized heating. The resulting surge of 1O2 and heat synergically destroyed membranes, dysfunction cellular self-defense and metabolic system, induced intracellular oxidative stress, and ultimately inactivated microorganisms. Additionally, the 1O2-dominated oxidation and cell adhesion facilitated the selective disinfection in real well water matrixes. This study provides a cost-effective and practical solution for efficient well water disinfection, which assists isolated rural areas in getting safe drinking water.

Co-exposure to highly allergenic airborne pollen and fungal spores in Europe.

The study is aimed at determining the potential spatiotemporal risk of the co-occurrence of airborne pollen and fungal spores high concentrations in different bio-climatic zones in Europe. Birch, grass, mugwort, ragweed, olive pollen and Alternaria and Cladosporium fungal spores were investigated at 16 sites in Europe, in 2005-2019. In Central and northern Europe, pollen and fungal spore seasons mainly overlap in June and July, while in South Europe, the highest pollen concentrations occur frequently outside of the spore seasons. In the coldest climate, no allergy thresholds were exceeded simultaneously by two spore or pollen taxa, while in the warmest climate most of the days with at least two pollen taxa exceeding threshold values were observed. The annual air temperature amplitude seems to be the main bioclimatic factor influencing the accumulation of days in which Alternaria and Cladosporium spores simultaneously exceed allergy thresholds. The phenomenon of co-occurrence of airborne allergen concentrations gets increasingly common in Europe and is proposed to be present on other continents, especially in temperate climate.

Inactivation of fungal spores by performic acid in water: Comparisons with peracetic acid.

Performic acid (PFA) has received increasing attention in water disinfection due to its high disinfection efficiency and fewer formation of disinfection by-products. However, the inactivation of fungal spores by PFA has not been investigated. In this study, the results showed that the log-linear regression plus tail model adequately described the inactivation kinetic of fungal spores with PFA. The k values of A. niger and A. flavus with PFA were 0.36 min-1 and 0.07 min-1, respectively. Compared to peracetic acid, PFA was more efficient in inactivating fungal spores and caused more serious damage on cell membrane. Compared to neutral and alkaline conditions, acidic environments demonstrated a greater inactivation efficiency for PFA. The increase of PFA dosage and temperature had a promoting effect on the inactivation efficiency of fungal spores. PFA could kill the fungal spores by damaging cell membrane and penetration of cell membranes. In real water, the inactivation efficiency declined as a result of the existence of background substances such as dissolved organic matter. Moreover, the regrowth potential of fungal spores in R2A medium were severely inhibited after inactivation. This study provides some information for PFA to control fungi pollution and explores the mechanism of PFA inactivation.

Passive fungal spore release from fruit and vegetable solid waste.

Food substrates in municipal solid wastes processing facilities and open dumpsites are a source for the release of fungal spores into air and can cause potential health and climate effects. Experiments were conducted in a laboratory scale flux chamber to measure the fungal growth and spore release from representative exposed cut fruit and vegetable substrates. The aerosolised spores were measured using an optical particle sizer. The results were compared to experiments conducted previously with a test species (Penicillium chrysogenum) on a synthetic media (czapek yeast extract agar). Significantly higher surface spore densities were observed for the fungi on the food substrates as compared to that on the synthetic media. The spore flux was high initially and then decreased on continued exposure to air. The spore emission flux normalised to the surface spore densities indicated that the emission from the food substrates was lower than the emissions from the synthetic media. A mathematical model was applied to the experimental data and the observed flux trends were explained in terms of the model parameters. A simple application of the data and the model to release from a municipal solid waste dumpsite was shown.

Kinetics and mechanism of sulfate radical-and hydroxyl radical-induced disinfection of bacteria and fungal spores by transition metal ions-activated peroxymonosulfate.

Peroxymonosulfate(PMS)-based advanced oxidation process have been recognized as efficient disinfection processes. This study comprehensively investigated the role of sulfate radical (SO4•-) and hydroxyl radical (•OH)-driven disinfection of bacteria and fungal spores by the PMS/metals ions (Me(II)) systems and modeled the CT value based on the relationship between survival and ∫[Radical]dt, with the aim to provide an accurate and quantitative kinetic data of inactivation processes. The results indicated that •OH played a more central role than SO4•- in the inactivation process, and bacteria were more vulnerable to radical attack than fungal spores due to the differences in antioxidant mechanisms and external structures. The k value of •OH -induced inactivation of E. coli was approximately 3-fold higher than that of A. niger, and the shoulder length of •OH -induced inactivation of E. coli was closely 52-fold shorter than that of A. niger after treated with the PMS/Co(II) system. The morphological and biochemical changes revealed that PMS/Me(II) treatment caused membrane damage, intracellular ROS accumulation and esterase activity loss in microorganisms. This study significantly improved the understanding of the contribution of radicals in the process of microbial inactivation by PMS/Me(II) and would provide important implications for the further development of technologies to cope with the highly resistant fungal spores in drinking water.

Aspergillus Conidia and Allergens in Outdoor Environment: A Health Hazard?

Aspergillus is a genus of saprophytic fungus widely distributed in the environment and associated with soil, decaying vegetation, or seeds. However, some species, such as A. fumigatus, are considered opportunistic pathogens in humans. Their conidia (asexual spores) and mycelia are associated with clinical diseases known as invasive aspergillosis (IA), mainly related to the respiratory tract, such as allergic asthma, allergic bronchopulmonary aspergillosis (ABPA), or hypersensitivity. However, they can also disseminate to other organs, particularly the central nervous system. Due to the dispersal mechanism of the conidia through the air, airborne fungal particle measurement should be used to prevent and control this mold. This study aims to measure the outdoor airborne concentration of Aspergillus conidia and the Asp f 1 allergen concentration in Bellaterra (Barcelona, Spain) during 2021 and 2022, and to compare their dynamics to improve the understanding of the biology of this genus and contribute to a better diagnosis, prevention, and therapeutic measures in the face of possible health problems. The results show that both particles were airborne nearly all year round, but their concentrations showed no correlation. Due to Asp f 1 not being present in the conidia itself but being detectable during their germination and in hyphal fragments, we report the relevance of the aero-immunological analysis as a methodology to detect the potential pathogenic hazard of this fungus.

Historical anthropogenic disturbances explain long-term moorland vegetation dynamics.

Upland moorlands are important landscapes, but many are considered degraded as a result of human activities. Consequently, their protection and restoration are of substantial concern. In Europe, restoration activities are often aimed at reversing the effects of 19th and 20th century "agricultural improvements," which often involved major drainage schemes. However, the ecological effects and long-term ecological context of "agricultural improvement" are not yet fully understood. To develop this understanding, we analyze paleoecological data (pollen, coprophilous fungal spores, microcharcoal) from five upland peatland sites using a range of analytical approaches: cluster analysis, principal component analysis, rate-of-change analysis, and regression analyses incorporating documentary historical data. The sites are located on Exmoor (South West England, UK), a landscape that typifies historic upland degradation. We demonstrate that in this landscape, 19th century drainage is associated with declines in Sphagnum and non-arboreal taxon richness; over longer timescales burning is associated with enhanced graminoid monocot abundance and grazing with lower taxon richness. We also show that rate-of-change in moorland vegetation communities during the 19th century is not distinctive in a long-term context: change has been a constant in this landscape, rather than an exception during the 19th century. Our findings indicate that the aims of "restoration" interventions intended to increase Sphagnum abundances, increase taxon richness and reduce graminoid dominance are consistent with the long-term dynamics of peatland systems, such as those on Exmoor. "Restoration" deemed successful in these terms may or may not resemble pre-drainage conditions, which were themselves a function of millennia of successive moorland management regimes.

Detection of Rice Fungal Spores Based on Micro- Hyperspectral and Microfluidic Techniques.

As rice is one of the world's most important food crops, protecting it from fungal diseases is very important for agricultural production. At present, it is difficult to diagnose rice fungal diseases at an early stage using relevant technologies, and there are a lack of rapid detection methods. This study proposes a microfluidic chip-based method combined with microscopic hyperspectral detection of rice fungal disease spores. First, a microfluidic chip with a dual inlet and three-stage structure was designed to separate and enrich Magnaporthe grisea spores and Ustilaginoidea virens spores in air. Then, the microscopic hyperspectral instrument was used to collect the hyperspectral data of the fungal disease spores in the enrichment area, and the competitive adaptive reweighting algorithm (CARS) was used to screen the characteristic bands of the spectral data collected from the spores of the two fungal diseases. Finally, the support vector machine (SVM) and convolutional neural network (CNN) were used to build the full-band classification model and the CARS filtered characteristic wavelength classification model, respectively. The results showed that the actual enrichment efficiency of the microfluidic chip designed in this study on Magnaporthe grisea spores and Ustilaginoidea virens spores was 82.67% and 80.70%, respectively. In the established model, the CARS-CNN classification model is the best for the classification of Magnaporthe grisea spores and Ustilaginoidea virens spores, and its F1-core index can reach 0.960 and 0.949, respectively. This study can effectively isolate and enrich Magnaporthe grisea spores and Ustilaginoidea virens spores, providing new methods and ideas for early detection of rice fungal disease spores.

Airborne pollen and fungi indoors: Evidence from primary schools in Lithuania.

The number of children suffering from respiratory allergies and asthma has been increasing worldwide and, hence, it is crucial to understand the burden of inhalant biological particles present in school facilities, where children spend one third of their life. From the perspective of indoor air quality, while there are numerous studies on outdoor bioaerosol exposure, there are still uncertainties regarding the diversity and deposition of airborne pollen and fungi indoors. When it comes to schools, there is limited research as to the potential bioaerosol exposure. Here we studied the indoor environment of public schools aiming to reveal whether primary schools of different sizes and at localities of different levels of urbanization may exhibit a variability in the biodiversity and abundance of particles of biological origin, which could pose a risk to child health. To achieve this, 11 schools were selected, located in a variety of environments, from downtown, to city centre-periphery, and to the suburbs. Fungal and pollen samples were collected from various surfaces in school classrooms and corridors, using passive air sampling and swab sampling. We demonstrated that fungi and pollen are detected in school premises during and after the vegetation season. The highest diversity of bioaerosols was found on the top of cabinets and windowsills, with Penicillium, Cladosporium and Acremonium being the most abundant indoors. The levels of fungi were higher in schools with more students. The diversity and amount of pollen in the spring were significantly higher than in samples collected in autumn. Our findings complemented existing evidence that bioaerosol measurements in schools (including kindergartens or informal education facilities) are vital. Hence, we here suggest that, in addition to monitoring air quality and bacterial levels indoors, fungi and pollen measurements have to be integrated in the existing regular biomonitoring campaigns so as to prevent exposure, increase awareness and manage efficiently allergic symptomatology.

Pre-exposure of peracetic acid enhances its subsequent combination with ultraviolet for the inactivation of fungal spores: Efficiency, mechanisms, and implications.

Waterborne fungi pose a potential threat to water supply safety due to their high resistance to disinfectants. Peracetic acid, as a promising alternative disinfectant to chlorine, has attracted increasing attention in water treatment. In this study, the inactivation of two dominant fungal species (Aspergillus niger and Aspergillus flavus) by sequential application of peracetic acid and ultraviolet (PAA-UV/PAA) was reported for the first time. Results revealed that the pre-exposure of PAA could facilitate the subsequent process of UV/PAA combination and shorten the lag phase in fungi inactivation. After 10 min of PAA pre-exposure, PAA-UV/PAA achieved 3.03 and 2.40 log inactivation of Aspergillus niger and Aspergillus flavus, which were 2- and 4.3-fold higher than that of direct UV/PAA under the same UV and PAA doses. PAA-UV/PAA disinfection also exhibited a stronger regrowth inhibition for incompletely inactivated fungal spores than direct UV/PAA. The increase of pH (5.0-9.0) and humic acid concentration (1.0-5.0 mg L - 1) showed an inhibitory effect on PAA-UV/PAA inactivation, but PAA-UV/PAA was more adaptable in a wide pH range and the presence of humic acid compared to direct UV/PAA. The more severe cell membrane damage and higher reactive oxygen species level in PAA-UV/PAA were evidenced for the first time by flow cytometry. The increased hydroxyl radical generation and higher synergism were primarily responsible for inactivation improvement. This study enhances the further understanding of the PAA-UV/PAA process, and the findings are expected to promote the development of PAA as a promising disinfectant for effective fungi control.