Potential Fungal Pathogens Research Articles

Black pepper pathogen suppression: Divergent rhizosphere fungal communities of healthy and diseased plants yield new insights for orchard management in Vietnam

Complex interactions involving soil physicochemical parameters and plant-associated microbial communities determine crop health. In Vietnam, this process is poorly understood in the context of black pepper production. Specifically, there is a dearth of information for improving the suppression of pathogenic fungi. Understanding the environmental dynamics influencing the distribution of these pathogens would facilitate the development and use of biological agents in black pepper pathogen management. Here, the molecular profiles of fungal communities from the rhizosphere of healthy and unhealthy Vietnamese black pepper orchards and their relationships were determined. Additionally, co-occurrence analyses with a previously constructed bacterial dataset identified taxa indicative of soil suppression. Alpha diversity of total fungi was influenced by only environmental factors, while that of arbuscular mycorrhizal fungi was more responsive to orchard health state. Glomus sp., Rhizophagus sp., Purpureocillium sp. and Plectosphaerella sp. were the most responsive genera to orchard health state. Potential fungal pathogens were more prevalent in the unhealthy orchards. Co-occurrence network analyses revealed that unhealthy orchards were less connected, had longer path distance and were missing putative pathogen-to-biocontrol interactions common in the healthy orchards. Soil electrical conductivity and potassium may be key factors in differentiating fungal communities of unhealthy from healthy orchards. This work highlights important microbial species and environmental considerations critical to improved black pepper management strategies.

Characterizing fungal communities on Mojave desert tortoises (Gopherus agassizii) in Arizona to uncover potential pathogens

The Mojave Desert tortoise (Gopherus agassizii) is a threatened reptile endemic to the southwestern United States. There are many factors contributing to its population decline such as human-driven habitat destruction and infectious diseases, which might include emerging fungal pathogens. We analyzed the fungal communities of two tortoises from northwest Arizona and compared the host mycobiome to the burrow mycobiome. Our main goal was to characterize the fungal community living on the Mojave Desert tortoise and to determine whether there are differences in community composition and potential pathogen abundance between the soil from burrows and tortoises’ mycobiome. Dissimilarities in communities would indicate that there are fungi adapted to the animal niche that are rarely present in soil and vice versa. Metagenomic analysis of ITS2 amplicons revealed significant differences in fungal communities between the substrate and samples taken from the animals. The tortoise samples show higher alpha diversity than the soil and are distinct from soil samples based on beta diversity analysis. We observed significantly greater relative abundance of potential fungal pathogens from the order Onygenales in the tortoise samples than the soil. These mycobiome data suggest that there is a unique fungal community occupying the desert tortoise host niche and this community contains a higher relative abundance of potential fungal pathogens. We argue that these fungi may pose a risk for increased morbidity and mortality among desert tortoises, which can in turn drive population decline.

Microbial diversity and potential health risks of household municipal solid waste in China: A case study in winter during outbreak of COVID-19

Microbial (bacteria and fungi) community structures and their distributions in urban household municipal solid waste (HMSW) were characterized in a sampling campaign in 38 cities of China covering 5 climatic zones. All samples were collected from garbage containers in residential communities during the Winter of 2022, from January 11 to 26. A total of 247 bacterial genera belonging to 22 phyla were identified among the samples. Firmicutes (44.3 %), Bacteroidetes (33.77 %) and Proteobacteria (21.54 %) were the top 3 dominant phyla, and Arcicella (33.11 %) and Leuconostoc (21.87 %) were the dominant genera. Meanwhile, 124 fungal genera from 7 fungal phyla were detected. Ascomycota was the most dominant phylum, with an average relative abundance of 77.31 %. Hanseniaspora (24.03 %), Debaryomyces (13.47 %), Candida (12.18) were the top 3 dominant fungal genera. Alpha-diversity analysis showed that the species richness and diversity of bacterial and fungal communities of HMSW samples belonging to different climatic zones did not differ significantly. Nonmetric multidimensional scaling (NMDS) analysis confirmed that climatic had an effect on microbial communities but did not show a significant correlation. In addition, the distribution of microbial community in different samples from the same climate zone varied considerably, suggesting the HMSW source play important role in shaping microbial community composition. Considering that residential HMSW is relatively fresh, we speculates that the original microorganisms residing in different components of HMSW are key influencing factor for the community, while the reshaping force driven by environmental conditions are relatively weak. In addition, the study identified 13 bacterial and 16 fungal pathogens with Pseudomonas putida (0.25 %) and Sclerotinia sclerotiorum (2.12 %) as the most abundant potential pathogenic bacteria and fungi, respectively. These findings provide valuable information for characterizing microbial features and potential risks of HMSW in its management system.

Effects of postharvest chitosan and potassium sorbate coating on the storage quality and fungal community of fresh jujube

Fresh jujube fruit are confronted with serious quality deterioration after harvest. This study explored the effects of chitosan and potassium sorbate coatings on the physicochemical quality and fungal community of jujube during cold storage. Microscopic observation showed that chitosan can uniformly distribute potassium sorbate on the surface of jujube. Chitosan coating inhibited the increase in respiration rate and total soluble solids and the decrease in titratable acidity and firmness. Jujube fruit treated with chitosan and potassium sorbate showed the lowest decay rate (2.67%) after 35 d of storage. High-throughput sequencing analysis indicated that the largest number of fungal amplicon sequence variants (ASVs) was detected in jujube treated with chitosan and potassium sorbate, exhibiting higher Chao1, Shannon and Simpson diversity indices. Treatment with chitosan or potassium sorbate alone reduced the relative abundance of Cladosporium in jujube fruit. Jujube treated with chitosan and potassium sorbate had the lowest total relative abundance of all potential pathogenic fungi (Cladosporium, Alternaria and Diaporthe). FUNGuild analysis showed that chitosan and potassium sorbate treatment could inhibit the relative abundance of plant pathogenic fungi. As a result, chitosan maintained the physicochemical properties of jujube by inhibiting the respiration rate, and the combination of chitosan and potassium sorbate reduced the decay rate of jujube by inhibiting pathogenic fungi. Therefore, chitosan and potassium sorbate coating is a promising approach to improve the storage quality and delay the decay of jujube fruit after harvest.

Work in nursing homes and occupational exposure to endotoxin and bacterial and fungal species.

Indoor microbial exposure may cause negative health effects. Only little is known about the occupational microbial exposure in nursing homes and the factors that influence the exposure. The exposure in nursing homes may be increased due to close contact with elderly persons who may carry infectious or antimicrobial-resistant microorganisms and due to handling of laundry, such as used clothing and bed linen. We investigated the microbial exposure in 5 nursing homes in Denmark, by use of personal bioaerosol samples from different groups of staff members taken during a typical working day, stationary bioaerosol measurements taken during various work tasks, sedimented dust samples, environmental surface swabs, and swabs from staff members' hands. From the samples, we explored bacterial and fungal concentrations and species composition, endotoxin levels, and antimicrobial resistance in Aspergillus fumigatus isolates. Microbial concentrations from personal exposure samples differed among professions, and geometric means (GM) were 2,159 cfu/m3 (84 to 1.5 × 105) for bacteria incubated on nutrient agar, 1,745 cfu/m3 (82 to 2.0 × 104) for bacteria cultivated on a Staphylococcus selective agar, and 16 cfu/m3 air for potential pathogenic fungi incubated at 37 °C (below detection limit to 257). Bacterial exposures were elevated during bed making. On surfaces, the highest bacterial concentrations were found on bed railings. The majority of bacterial species found were related to the human skin microflora, such as different Staphylococcus and Corynebacterium species. Endotoxin levels ranged from 0.02 to 59.0 EU/m3, with a GM of 1.5 EU/m3. Of 40 tested A. fumigatus isolates, we found one multiresistant isolate, which was resistant towards both itraconazole and voriconazole, and one isolate resistant towards amphotericin B. In conclusion, we give an overview of the general microbial exposure in nursing homes and show that microbial exposures are higher for staff with more care and nursing tasks compared with administrative staff.

Soil amendments impact root‐associated fungal communities of balsam poplar on a phosphogypsum reclamation site

During reclamation organisms like fungi directly interact at the soil–plant interface and can potentially impact plant growth, health, and overall revegetation success. Therefore, a study was conducted to determine how topsoil amendments (mycorrhizae, biochar, compost, manure) influence root‐associated fungal communities and growth of fast growing Populus balsamifera trees on a phosphogypsum (PG) reclamation site in Alberta, Canada. Tree height, diameter, biomass, and diversity of Ascomycota and Basidiomycota were assessed using Illumina MiSeq sequencing targeting the fungal internal transcribed spacer region 1 (ITS1) and ITS2. Topsoil amendments and root location (topsoil vs. PG within a treatment), greatly affected the root fungal community. Basidiomycota preferentially grew in PG, and had a high treatment specific root fungal community with highest diversity of ectomycorrhizal fungi and basidiomycete yeast in biochar PG and compost PG. Populus balsamifera roots were associated with a high diversity of Ascomycota, found equally in roots in topsoil and PG, with a trend towards higher diversity and abundance in PG. The core genus found across all samples was Fusarium, a potential plant pathogen. All topsoil amendments strongly reduced abundance of Fusarium and other Ascomycota in poplar roots. Biochar and compost reduced number and abundance of potential fungal pathogens and increased number of potential plant growth promoting Basidiomycota, indicating favorable growth conditions for P. balsamifera on a phosphygypsum reclamation site by application of these amendments. This study indicates compost and biochar can be effective soil amendments to support P. balsamifera establishment and growth on a PG reclamation site.

Red clover root-associated microbiota is shaped by geographic location and choice of farming system.

This study evaluated the red clover (Trifolium pratense) root-associated microbiota to clarify the presence of pathogenic and beneficial microorganisms in 89 Swedish field sites. 16S rRNA and ITS amplicon sequencing analysis were performed on DNA extracted from the red clover root samples collected to determine the composition of the prokaryotic and eukaryotic root-associated microbe communities. Alpha and beta diversities were calculated and relative abundance of various microbial taxa and their co-occurrence were analyzed. Rhizobium was the most prevalent bacterial genus, followed by Sphingomonas, Mucilaginibacter, Flavobacterium, and the unclassified Chloroflexi group KD4-96. The Leptodontidium, Cladosporium, Clonostachys, and Tetracladium fungal genera known for endophytic, saprotrophic, and mycoparasitic lifestyles were also frequently observed in all samples. Sixty-two potential pathogenic fungi were identified with a bias toward grass pathogens and a higher abundance in samples from conventional farms. We showed that the microbial community was mainly shaped by geographic location and management procedures. Co-occurrence networks revealed that the Rhizobiumleguminosarum bv. trifolii was negatively associated with all fungal pathogenic taxa recognized in this study.

Biogas Residues Improved Microbial Diversity and Disease Suppression Function under Extent Indigenous Soil Microbial Biomass.

Indigenous soil microbial biomass (ISMB) plays a key role in maintaining essential functions and biodiversity of soil health. One of the critical unknowns is how the indigenous microorganisms respond to different fertilizers which is directly related to agricultural production. Therefore, we used Mi-Seq sequencing and network analyses to compare the response of ISMB to biogas residue and chemical fertilizers. The results showed that crop production was profoundly influenced by levels of ISMB present and is further dependent on the strategy of fertilizer application. Higher ISMB primarily manifests through retention of richer microbial abundance, a balanced community structure, and tightened co-occurrence within a certain proportion of Nitrospirae, Rhizophlyctidaceae, and Gemmatimonadetes. Compared to chemical fertilizer, biogas residue resulted in higher production with more strongly linked nodes such as Actinobacteria, Chloroflexi and Gemmatimonadetes. Under the same level of ISMB, the microbial diversity was richer and co-occurrence was tighter when biogas residues were applied compared with chemical fertilizer. In addition, the higher level of ISMB with biogas residue applied had a lower abundance of potential fungal pathogens in both bulk and rhizosphere soil compared with chemical fertilizer. This study provides critical data to understand the influence of ISMB and biogas residue on soil ecological system.

The Potential Fungal Pathogens of Euonymus japonicus in Beijing, China.

Euonymus japonicus tolerates the dry and frigid climate of Beijing, China, and effectively filters out particles during the winter. However, fungal infestation frequently causes extreme illness and can even lead to shrub death. In this study, 104 diseased E. japonicus specimens were collected from seven districts in Beijing. Seventy-nine isolates were identified as 22 fungal species in seven genera. The species were Aplosporella hesperidica, A. javeedii, A. prunicola, Botryosphaeria dothidea, Colletotrichum aenigma, Co. euonymi, Co. euonymicola, Co. gloeosporioides, Cytospora ailanthicola, C. albodisca, C. diopuiensis, C. discotoma, C. elaeagni, C. euonymicola, C. euonymina, C. haidianensis, C. leucostoma, C. sophorae, C. zhaitangensis, Diaporthe eres, Dothiorella acericola, and Pestalotiopsis chaoyangensis. On the basis of morphological and phylogenetic analyses, Colletotrichum euonymi, Co. euonymicola, Cytospora zhaitangensis, and Pestalotiopsis chaoyangensis were introduced as novel species. Colletotrichum euonymi, Co. euonymicola, and Pestalotiopsis chaoyangensis were subsequently confirmed as pathogens of E. japonicus leaves by pathogenicity testing. This study provides an important assessment of the fungi associated with diseases of E. japonicus in Beijing, China.

Caspian Sea Mycosands: The Variety and Abundance of Medically Important Fungi in Beach Sand and Water.

Samples from a total of 67 stations, distributed amongst 32 cities along the Caspian Sea coastline, were collected during the summer of 2021 on sunny days. The samples were collected from each station, including both dry/wet sand and shoreline water. The grown samples were primarily analyzed for the macro/microscopic morphologic features of the fungi. Moreover, identification by PCR-RFLP was performed for yeasts, dermatophytes, and Aspergillus sp. strains. Antifungal susceptibility tests were performed for probable-isolated Aspergillus and Candida sp. A total of 268 samples were collected, from which 181 (67.54%) isolates were recovered. Yeast-like fungi and potential pathogenic black fungi were detected in 12 (6.6%) and 20 (11%) of the sand (dry/wet) samples. Potential pathogenic hyaline fungi were identified in 136 (75.1%) samples, in which Aspergillus sp. was the predominant genus and was detected in 76/136 (47.8%) samples as follows: A. section Flavi n = 44/76 (57.9%), A. section Nigri n = 19/76 (25%), A. section Nidulantes n = 9/76 (11.8%), and A. section Fumigati n = 4/76 (5.3%). The most effective azole antifungal agent was different per section: in A. section Fumigati, PSZ; in Aspergillus section Nigri, ITZ and ISZ; in A. section Flavi, EFZ; and in A. section Nidulantes, ISZ. Candida isolates were susceptible to the antifungals tested.

Shaping effects of rice, wheat, maize, and soybean seedlings on their rhizosphere microbial community.

The rhizosphere microbiome plays critical roles in plant growth and is an important interface for resource exchange between plants and the soil environment. Crops at various growing stages, especially the seedling stage, have strong shaping effects on the rhizosphere microbial community, and such community reconstruction will positively feed back to the plant growth. In the present study, we analyzed the variations of bacterial and fungal communities in the rhizosphere of four crop species: rice, soybean, maize, and wheat during successive cultivations (three repeats for the seedling stages) using 16S rRNA gene and internal transcribed spacer (ITS) high-throughput sequencing. We found that the relative abundances of specific microorganisms decreased after different cultivation times, e.g., Sphingomonas, Pseudomonas, Rhodanobacter, and Caulobacter, which have been reported as plant-growth beneficial bacteria. The relative abundances of potential plant pathogenic fungi Myrothecium and Ascochyta increased with the successive cultivation times. The co-occurrence network analysis showed that the bacterial and fungal communities under maize were much more stable than those under rice, soybean, and wheat. The present study explored the characteristics of bacteria and fungi in crop seedling rhizosphere and indicated that the characteristics of indigenous soil flora might determine the plant growth status. Further study will focus on the use of the critical microorganisms to control the growth and yield of specific crops.

Effects of micro-positive pressure environment on nitrogen conservation and humification enhancement during functional membrane-covered aerobic composting

Aerobic composting is a humification process accompanied by nitrogen loss. This study is the first research systematically investigating and elucidating the mechanism by which functional membrane-covered aerobic composting (FMCAC) reduces nitrogen loss and enhances humification. The variations in bioavailable organic nitrogen (BON) and humic substances (HSs) in different composting systems were quantitatively studied, and the functional succession patterns of fungal groups were determined by high-throughput sequencing and FUNGuild. The FMCAC improved oxygen utilization and pile temperature, increased BON by 29.95 %, reduced nitrogen loss by 34.00 %, and enhanced humification by 26.09 %. Meanwhile, the FMCAC increased the competitive advantage of undefined saprotroph and significantly reduced potential pathogenic fungi (<0.10 %). Structural equation modeling indicated that undefined saprotroph facilitated the humification process by increasing the production of BON and storing BON in stable humic acid. Overall, the FMCAC increased the safety, stability, and quality of the final compost product.

Microbiome analysis and biocontrol bacteria isolation from rhizosphere soils associated with different sugarcane root rot severity.

To explore the causal pathogen and the correlated rhizosphere soil microecology of sugarcane root rot, we sampled the sugarcane root materials displaying different disease severity, and the corresponding rhizosphere soil, for systematic root phenotype and microbial population analyses. We found that with increased level of disease severity reflected by above-ground parts of sugarcane, the total root length, total root surface area and total volume were significantly reduced, accompanied with changes in the microbial population diversity and structure in rhizosphere soil. Fungal community richness was significantly lower in the rhizosphere soil samples from mildly diseased plant than that from either healthy plant, or severely diseased plant. Particularly, we noticed that a peculiar decrease of potential pathogenic fungi in rhizosphere soil, including genera Fusarium, Talaromyces and Neocosmospora, with increased level of disease severity. As for bacterial community, Firmicutes was found to be of the highest level, while Acidobacteria and Chloroflexi of the lowest level, in rhizosphere soil from healthy plant compared to that from diseased plant of different severity. FUNGuild prediction showed that the proportion of saprophytic fungi was higher in the rhizosphere soil of healthy plants, while the proportion of pathogenic fungi was higher in the rhizosphere soil of diseased plants. By co-occurrence network analysis we demonstrated the Bacillus and Burkholderia were in a strong interaction with Fusarium pathogen(s). Consistently, the biocontrol and/or growth-promoting bacteria isolated from the rhizosphere soil were mostly (6 out of 7) belonging to Bacillus and Burkholderia species. By confrontation culture and pot experiments, we verified the biocontrol and/or growth-promoting property of the isolated bacterial strains. Overall, we demonstrated a clear correlation between sugarcane root rot severity and rhizosphere soil microbiome composition and function, and identified several promising biocontrol bacteria strains with strong disease suppression effect and growth-promoting properties.

Reducing plant pathogens could increase crop yields after plastic film mulching

Soil fungi are closely associated with crop growth in agricultural ecosystems through processes such as nutrient uptake and pathogenesis. Plastic film mulching (PM) plays a dominant role in increasing crop yields in dryland agriculture worldwide. The functional guilds of soil fungi under PM and their effects on crops remain unclear. In this study, we explored the absolute abundance, diversity, community composition, and functional guilds of soil fungi after short-term (2 years) and long-term (10 years) mulching experiments. Short-term mulching caused a 37 %–51 % decrease in absolute fungal abundance owing to abrupt changes in the microenvironment. The response of the fungal community to PM varied with sites, with the effect being more pronounced under poor hydrothermal conditions (314 mm). The abundance of potential fungal pathogens decreased under PM; for example, Gibberella (maize ear rot) abundance was 45 % and 72 % lower under short- and long-term mulching, respectively, when compared with that in control. In contrast, the abundance of plant biocontrol fungi increased under PM; for instance, Glomeromycota abundance increased twofold under long-term mulching. Although PM did not alter the complexity and stability of fungal co-occurrence network, competition among fungi increased in the absence of sufficient carbon (C) sources. Long-term mulching reduced phytopathogen guilds by 12 %–77 % and increased arbuscular mycorrhizal fungi (AMF) guilds by 89 %–94 %. Structural equation modeling suggested that PM altered fungal functional guilds mainly by shaping the structure of the fungal community, and fungal pathogens decreased with increased AMF functional guilds, inducing higher maize yields. These results showed for the first time, from a microbial perspective, that pathogens reduction owing to PM could explain 4.4 % of maize yield variation, providing theoretical guidance to accomplish sustainability of continuous maize mulching.

Sapwood mycobiome varies across host, plant compartment and environments in Nothofagus forests from Northern Patagonia.

Global forests are increasingly being threatened by altered climatic conditions and increased attacks by pests and pathogens. The complex ecological interactions among pathogens, microbial communities, tree hosts and the environment are important drivers of forest dynamics. Little is known about the ecology of forest pathology and related microbial communities in temperate forests of the southern hemisphere. In this study, we used next-generation sequencing to characterize sapwood-inhabiting fungal communities in North Patagonian Nothofagus forests and assessed patterns of diversity of taxa and ecological guilds across climatic, site and host variables (health condition and compartment) as a contribution to Nothofagus autecology. The diversity patterns inferred through the metabarcoding analysis were similar to those obtained through culture-dependent approaches. However, we detected additional heterogeneity and greater richness with culture-free methods. Host species was the strongest driver of fungal community structure and composition, while host health status was the weakest. The relative impacts of site, season, plant compartment and health status were different for each tree species; these differences can be interpreted as a matter of water availability. For Nothofagus dombeyi, which is distributed across a wide range of climatic conditions, site was the strongest driver of community composition. The microbiome of N. pumilio varied more with season and temperature, a relevant factor for forest conservation in the present climate change scenario. Both species carry a number of potential fungal pathogens in their sapwood, whether they exhibit symptoms or not. Our results provide insight into the diversity of fungi associated with the complex pathobiome of the dominant Nothofagus species in southern South America.

The Effect of Different Remediation Treatments on Soil Fungal Communities in Rare Earth Tailings Soil

Extensive mining of rare earth deposits has caused severe soil erosion, resulting in the degradation of plant–soil systems and the reduction in microbial diversity. Combined ecological remediation technology is the key method of vegetation reconstruction and ecological restoration in abandoned tailings. In this study, the effects of different cover crops–biochar–organic fertilizer and biochar–organic fertilizer treatments on soil fungal communities in rare earth tailings soil were analysed using high-throughput sequencing technology. Linear discriminant analysis effect size (LEfSe) was used to analyse saprophytic, mycorrhizal, and potential pathogenic fungi in soils after different combined remediations. Moreover, the effects of soil environmental factors on fungal community species’ composition were analysed by redundancy analysis (RDA) and variance partitioning analysis (VPA) after different combined remediations. LEfSe indicated a risk of citrus pathogenicity by Diaporthaceae indicator fungi after biochar–organic fertilizer combined treatment. RDA and VPA revealed that pH was the main environmental factor affecting the fungal community in the different combined remediation treatments. Additionally, the Paspalum wettsteinii cover crops–biochar–organic fertilizer and biochar–livestock manure treatments were more conducive to arbuscular mycorrhizal fungi recruitment. We also clarified the fungal community composition structure, soil environmental factors, and fungal community relationships in rare earth tailings soil after different combined remediation treatments.

Domestic refrigerators: An overlooked breeding ground of antibiotic resistance genes and pathogens

Domestic refrigerator is a widely used appliance to keep food fresh and retard food spoilage in household. However, our understanding of microbial health risk associated with food under such circumstance still remains very poor. Here, typical types of food (vegetable, fish, and pork) were kept in a domestic refrigerator at 4 °C for 3–30 days. Temporal dynamics of antibiotic resistome, pathogens, bacterial and fungal communities during this period were investigated via high-throughput quantification and Illumina sequencing technologies. Results showed that a large number (21–134) of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) were detected across the three food types, including 10.06 % of high-risk ARGs classified by their risk ranks. Moreover, four bacterial pathogens (i.e., Bacillus cereus, Cronobacter spp., Klebsiella pneumoniae and Staphylococcus aureus) targeted by marker genes including the pathogen-specific genes or virulence factor genes, and some potential fungal pathogens (e.g., Fusarium, Candida, and Aspergillus) were detected, indicating the occurrence of microbial risk even at the normally regarded safe storage temperature. Among all food types, the total bacterial density and ARG abundances in fish rapidly increased after only 3 days, much faster than vegetable and pork after 10 days. In addition, fish samples contained the highest ARG and pathogen abundances, indicating its potentially higher health risk than other food types. Finally, the shifts of ARG pattern were mainly contributed by bacterial communities and MGEs. This study highlights that food preserved in refrigerator at 4 °C could still be an unneglected microbial risk, and raises awareness of improving food safety in domestic environment.

Molecular detection of Aspergillus in respiratory samples collected from patients at higher risk of chronic pulmonary aspergillosis

ObjectiveAspergillosis diagnosis depends on the detection of Aspergillus in biological samples ─ usually using cultural and immunoenzyme techniques ─ but their sensitivity and specificity varies. We aimed to study the prevalence of Aspergillus in patients at higher risk of chronic pulmonary aspergillosis (i.e., HIV-infected patients and individuals with active or previous tuberculosis), and to determine the potential role of molecular approaches to increase detection of Aspergillus in respiratory samples. MethodsThe DNA extracted from 43 respiratory samples that had been previously analyzed by immunoenzyme and/or cultural techniques was amplified by real-time multiplex PCR, and the results of these methods were compared. We also sequenced the ITS1 region and the calmodulin gene in 10 respiratory samples to perform a pilot metagenomic study to understand the ability of this methodology to detect potential pathogenic fungi in the lung mycobiome. ResultsReal-time Aspergillus PCR test exhibited a higher positivity rate than the conventional techniques used for aspergillosis diagnosis, particularly in individuals at risk for chronic pulmonary aspergillosis. The metagenomic analysis allowed for the detection of various potentially pathogenic fungi. ConclusionsMolecular techniques, including metagenomics, have great ability to detect potentially pathogenic fungi rapidly and efficiently in human biological samples.

Significant differences in intestinal fungal community of hooded cranes along the wintering periods.

The intestinal microbiota play vital roles for health of wild birds in many ways. Migratory birds with unique life history might increase the risk of pathogenic transmission across the regions. However, few studies have clarified the fungal community structure and inferred the potential pathogens in guts of migratory birds. The high-throughput sequencing method was applied to analyze the fungal community structure and detect the potential fungal pathogens in guts of hooded cranes among different wintering stages. Significant differences were found in gut fungal community composition of hooded cranes among three wintering stages, with the lowest fungal diversity in the late wintering stage. In the late stage, hooded cranes harbored higher relative abundance of plant saprotroph, contributing to food digestion for hosts. Hooded cranes were associated with the lowest diversity and relative abundance of animal pathogens in the late wintering stage. There was an increasing trend of deterministic process for gut fungal community assembly, suggesting that hosts interaction with their fungal communities changed by enhanced gut selection/filtering along wintering periods. Hooded crane was associated with the strongest gut selection/filtering to obtain defined gut fungal community with retaining probiotics (i.e., plant saprotroph) and exclusion of certain pathogens in the late wintering stage. Overall, these results demonstrated that hooded cranes might regulate their gut microbiota to enhance digestion and decrease gut pathogens in preparation for long-term migration.

Biogas slurry application alters soil properties, reshapes the soil microbial community, and alleviates root rot of Panax notoginseng.

BackgroundPanax notoginseng is an important herbal medicine in China, where this crop is cultivated by replanting of seedlings. Root rot disease threatens the sustainability of P. notoginseng cultivation. Water flooding (WF) is widely used to control numerous soilborne diseases, and biogas slurry shows positive effects on the soil physiochemical properties and microbial community structure and has the potential to suppress soilborne pathogens. Hence, biogas slurry flooding (BSF) may be an effective approach for alleviating root rot disease of P. notoginseng; however, the underlying mechanism needs to be elucidated.MethodsIn this study, we conducted a microcosm experiment to determine if BSF can reduce the abundance of pathogens in soil and, alleviate root rot of P. notoginseng. Microcosms, containing soil collected from a patch of P. notoginseng showing symptoms of root rot disease, were subjected to WF or BSF at two concentrations for two durations (15 and 30 days), after which the changes in their physicochemical properties were investigated. Culturable microorganisms and the root rot ratio were also estimated. We next compared changes in the microbial community structure of soils under BSF with changes in WF and untreated soils through high-throughput sequencing of bacterial 16S rRNA (16S) and fungal internal transcribed spacer (ITS) genes amplicon.ResultsWF treatment did not obviously change the soil microbiota. In contrast, BSF treatment significantly altered the physicochemical properties and reshaped the bacterial and fungal communities, reduced the relative abundance of potential fungal pathogens (Fusarium, Cylindrocarpon, Alternaria, and Phoma), and suppressed culturable fungi and Fusarium. The changes in the microbial community structure corresponded to decreased root rot ratios. The mechanisms of fungal pathogen suppression by BSF involved several factors, including inducing anaerobic/conductive conditions, altering the soil physicochemical properties, enriching the anaerobic and culturable bacteria, and increasing the phylogenetic relatedness of the bacterial community.ConclusionsBSF application can reshape the soil microbial community, reduce the abundance of potential pathogens, and alleviate root rot in P. notoginseng. Thus, it is a promising practice for controlling root rot disease in P. notoginseng.