Fungal Pathogens Research Articles

Comparative Analysis of Some Morpho-cultural Characters of Venturia inaequalis (Apple Scab Pathogen) under Kashmir Conditions

Apple (Malus × domestica Borkh.) represents one of the most widely cultivated and economically important temperate fruit crops in the world. However, its cultivation faces significant challenges due to various fungal pathogens, particularly apple scab inflicted by Venturia inaequalis, which has a profound impact on the yield and quality of apple, resulting in substantial economic loss. The pervasive nature of V. inaequalis and its ability to damage crops poses a serious threat to sustainable apple production. In this study, the morpho-cultural characteristics of V. inaequalis were systematically analyzed and compared between the Shopian and Baramullah districts of the Kashmir Valley. Ten isolates from each district were grown on Malt Extract Agar (MEA) and Potato Dextrose Agar (PDA) for comparative analysis. The colonies were characterized by a compact hue ranging from greyish-green to blackish and exhibited a densely velvety texture. Mycelial growth was predominantly aerial, with some isolates displaying partial irregularity in colony margins, while others exhibited greater uniformity. The mean colony radii for potato dextrose agar and malt extract agar media was measured as 23.01 mm and 28.50 mm for the Shopian district, and 28.09 mm and 33.33 mm for the Baramullah district, respectively. The mean conidial spore density was recorded at 27.87 spores/cm² for the Baramullah district and 34.67 spores/cm² for the district Shopian, respectively. This variation could be attributed to several factors, including genetic diversity within the pathogen populations, differences in altitude, climate, and other environmental factors, the selection pressure exerted by fungicide applications, the emergence of novel pathogen races, and phenotypic plasticity driven by the differing agricultural practices and microclimatic conditions in the respective regions.

First Report of Alternaria alternata Causing Alternaria Leaf Spot of Black Nightshade in China.

Black nightshade (Solanum nigrum L.) belongs to Solanaceae family, which is distributed all over China and has become one of the malignant weeds in cotton field in Xinjiang (Zhang et al., 2020; Zhang et al., 2021). During July 2016 to August 2023, leaf spot was observed on black nightshade in Hami and Shihezi city, Xinjiang, China. The spots appeared as brown circular or irregular, with some small black dots surrounded with yellow halos. Twenty five black nightshade plants were collected from a cotton field (100 m2) for pathogen isolation in Shihezi, July 2023. Leaf samples (2×2 mm) from the boundary between diseased tissue and healthy tissue, sterilized in 70% ethanol for 45 s, dipped in 2% NaClO for 30 s, rinsed three times with sterile distilled water, placed on potato dextrose agar (PDA) after dried and cultivated at 25°C in the dark for 4 days. Twenty-five fungal isolates were obtained and purified by using the single-spore isolation method, and all isolates were found to be similar in morphology, appeared as dark brown and produced dark brown pigmentation on PDA. Conidiophores were black brown, 8 to 58 × 10 to 20 μm, with 1 to 5 transverse septa and 0 to 3 longitudinal septa, the results similar to those of Alternaria alternata isolated from buckwheat (Li et al., 2021). To confirm this identification, total genomic DNA of the isolates were extracted by CTAB method (Doyle et al., 1987). The internal transcribed spacer (rDNA-ITS), Alternaria major allergen gene (Alt α1) and glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) of the A. alternata 26 were amplified using the primers ITS1/ITS4 (Glass et al., 1995), Altα1-F/Altα1-R (Li et al., 2024) and gpd-F/gpd-R (5'-CAACGGCTTCGGTCGCAT- TG-3' / 5'-GCCAAGCAGTTGGTTGTG-3'), respectively. The rDNA-ITS, Altα1 and GAPDH gene sequences were deposited in GenBank (KX904867, PP263361, PP263360), and showed 100% identity (rDNA-ITS: 572 out of 609 bp; Alt α1: 514 out of 516 bp; GAPDH: 619 out of 619 bp) to A. alternata (KU179665; MW522975; MK451977), respectively. The multi-gene phylogenetic tree showed that the representative isolate was grouped with A. alternata and identified as A. alternata. Black nightshade was used for pathogenicity assay. Fifteen plants at 3 to 4-leaf stage were selected and sprayed with the conidial suspension (1.0×106 spores/mL) of A. alternata 26 after wound the underside of leaves (2 mm length) with inoculating needle. Five plants were sprayed with sterilized water as control. After inoculation, placed the plants in a plastic box and covered with plastic wrap to keep 80% relative humidity for 3 days at 25℃ in the greenhouse, then removed the plastic box. Symptoms of leaf spots appeared within 12 - 15 days that were similar to the symptoms observed in the field, and the fungal pathogen that was re-isolated from symptomatic leaves was identical to original pathogen on the basis of morphological and molecular analysis to fulfill Koch's postulates. This experiment was repeated three times, and the disease incidence was above 80% after inoculation at each time. To our knowledge, this is the first report of A. alternata causing leaf spot of black nightshade in China. This report will help us to monitoring distribution of the disease and providing theoretical basis for disease prevention and control.

Infection by a fungal pathogen and mating behavior in Pacific treefrogs: a test of the Hamilton-Zuk hypothesis

Infection by a fungal pathogen and mating behavior in Pacific treefrogs: a test of the Hamilton-Zuk hypothesis

Cryptococcus neoformans infections: aspartyl protease potential to improve outcome in susceptible hosts.

Vaccination and immunotherapies against fungal pathogens still remain a challenge. Here, we show using an in vivo model based on outbred mice that development of antibodies against Pep1p, an antigenic protein of the fungal pathogen Cryptococcus neoformans, confers resistance to this fungal infection. In support of this observation, prophylactic or therapeutic immunization of the mice with recombinant Pep1p could improve their survival when infected with a lethal dose of C. neoformans. Moreover, passive therapy with monoclonal anti-Pep1p antibodies also enhanced survival of the mice from C. neoformans infection. The associated antifungal mechanisms were mounting of a protective immune response and the development of fungal specific antibodies that decrease the fungal burden due to an increase in their phagocytosis and/or inhibit the fungal multiplication. Together, our study demonstrates (a) the mode of host-fungal interaction and the immune response developed thereby play a crucial role in developing resistance against C. neoformans; (b) Pep1p, an aspartic protease as well as an antigenic protein secreted by C. neoformans, can be exploited for vaccination (both prophylactic and therapeutic) or immunotherapy to improve the host defense during this fungal infection.

Seeds Priming with Bio-Silver Nanoparticles Protects Pea (Pisum sativum L.) Seedlings Against Selected Fungal Pathogens

Nano-priming is a relatively new seed treatment technique using metal and metal oxide nanoparticles (NPs), and such application of NPs may support the plants' immunity. Recently we have shown that the that biologically synthesized silver nanoparticles (bio-AgNPs) used as short-term foliar treatment protect pea seedlings against D. pinodes and F. avenaceum. In the present study, the protection of peas against both fungal pathogens via seed priming with bio-AgNPs was analyzed. Moreover, the changes in the polar metabolic profiles of the seedlings caused by priming and infection were also compared. Seed priming with bio-AgNPs at concentrations of 50 and 100 mg/L considerably reduced the symptoms and infection levels of both pathogens by over 70% and 90% for F. avenaceum and D. pinodes, respectively. Pathogens infection and nano-priming affected the metabolic profile of pea seedlings. The major changes in the primary metabolism were observed among carbohydrates and amino acids. In turn, this may result in changes in the expression and accumulation of secondary metabolites. Therefore, further investigation of the effect of nano-priming should focus on the changes in the secondary metabolism.

On the structure of species-function participation in multilayer ecological networks

Understanding how biotic interactions shape ecosystems and impact their functioning, resilience and biodiversity has been a sustained research priority in ecology. Yet, traditional assessments of ecological complexity typically focus on species-species interactions that mediate a particular function (e.g., pollination), overlooking both the synergistic effect that multiple functions might develop as well as the resulting species-function participation patterns that emerge in ecosystems that harbor multiple ecological functions. Here we propose a mathematical framework that integrates various types of biotic interactions observed between different species. Its application to recently collected data of an islet ecosystem—reporting 1537 interactions between 691 plants, animals and fungi across six different functions (pollination, herbivory, seed dispersal, decomposition, nutrient uptake, and fungal pathogenicity)—unveils a non-random, nested structure in the way plant species participate across different functions. The framework further allows us to identify a ranking of species and functions, where woody shrubs and fungal decomposition emerge as keystone actors whose removal have a larger-than-random effect on secondary extinctions. The dual insight—from species and functional perspectives—offered by the framework opens the door to a richer quantification of ecosystem complexity and to better calibrate the influence of multifunctionality on ecosystem functioning and biodiversity.

Bacterial Allies in Agricultural Defense: Evaluating Xenorhabdus and Photorhabdus Supernatants Against Phytophthora infestans and Monilinia laxa

Fungal plant pathogens represent a significant threat to global agriculture, affecting crop productivity and food security. Phytophthora infestans and Monilinia laxa are two such pathogens causing blights and brown rot, respectively, in economically vital crops like potato and stone fruits. Developing sustainable management strategies is crucial to mitigate these threats. Xenorhabdus and Photorhabdus bacteria produce various secondary metabolites with different biological activities. This study investigates the antifungal activity of cell-free supernatants from Xenorhabdus and Photorhabdus bacteria against P. infestans and M. laxa. Results demonstrate varying degrees of antifungal efficacy among bacterial species, with X. cabanillasii and X. szentirmaii exhibiting significant suppression of fungal growth. The findings underscore the importance of exploring biocontrol agents in integrated pest management practices.

Candida glabrata maintains two HAP1 ohnologs, HAP1A and HAP1B, for distinct roles in ergosterol gene regulation to mediate sterol homeostasis under azole and hypoxic conditions.

Invasive and drug-resistant fungal infections pose a significant public health concern. Candida glabrata, a human fungal pathogen, is often difficult to treat due to its intrinsic resistance to azole antifungal drugs and its capacity to develop clinical drug resistance. Therefore, understanding the pathways that facilitate fungal growth and environmental adaptation may lead to novel drug targets and/or more efficacious antifungal therapies. While the mechanisms of azole resistance in Candida species have been extensively studied, the roles of zinc cluster transcription factors, such as Hap1A and Hap1B, in C. glabrata have remained largely unexplored until now. Our research shows that these factors play distinct yet crucial roles in regulating ergosterol homeostasis under azole drug treatment and oxygen-limiting growth conditions. These findings offer new insights into how this pathogen adapts to different environmental conditions and enhances our understanding of factors that alter drug susceptibility and/or resistance.

Identification and Characterization of Bipolaris secalis and B. setariae Associated with Brown Stripe Disease on Saccharum officinarum in Yunnan, China.

Sugarcane (Saccharum officinarum L.) is the most important sugar crop that belongs to the Poaceae family and is mainly cultivated in tropical and subtropical regions worldwide. Brown stripe disease is a common and serious foliar fungal disease of sugarcane and has become a serious threat to sugarcane production in China. Yunnan Province is the second-largest sugar base in China. From 2022 to 2023, brown stripe-like symptoms from different sugarcane variety were observed in Yunnan. Thirty-nine Bipolaris-like isolates were obtained from symptomatic leaves of different sugarcane varieties and characterized by means of morphological identification combined with multilocus phylogeny comprising internal transcribed spacer rDNA (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the translation elongation factor-1α (TEF-1α) gene sequence date. The analysis provided strong support for the delineation and identification of two Bipolaris species associated with sugarcane brown stripe, including Bipolaris secalis and B. setariae. Pathogenicity text in potted sugarcane plants confirmed that these two Bipolaris species can cause brown stripe of sugarcane, thus fulfilling Koch's postulates. This study confirmed that B. secalis and B. setariae were the pathogens of brown stripe in Yunnan Province. To the best of our knowledge, this is the first to report B. secalis as causal agents of brown stripe of sugarcane. Collectively, this finding provides a basis for sugarcane brown stripe disease accurate diagnosis and would be helpful to the development of effective management strategies.

Fludioxonil does not antagonize propiconazole activity against Geotrichum candidum.

Fludioxonil and propiconazole are frequently used fungicides for managing fungal diseases in specialty crops. They are often used in mixture to manage brown rot caused by Monilinia fructicola and sour rot caused by the yeast-like fungus Geotrichum candidum,respectively, on stone fruits. Research indicated, however, that fludioxonil can increase the expression of the ATP-binding cassette transporters CDR1 and CDR2 in the yeast Candida albicans, leading to increased resistance to DMI fungicides. In this study, mycelial growth assays and detached fruit assays were conducted with G. candidum isolates of varying sensitivity to propiconazole to investigate potential antagonism between the two active ingredients. Isolates sensitive, reduced-sensitive, and resistant to propiconazole were able to grow with or without 20 µg/ml fludioxonil on PDA containing up to 0, 3, and 30 μg/ml propiconazole and on water agar containing up to 7.5, 7.5, and 30 μg/ml propiconazole, respectively, and no antagonistic interactions were detected. Propiconazole controlled disease with or without 20 µg/ml fludioxonil equally on detached cherry fruit caused by isolates with different DMI sensitivity phenotypes. Our results indicated no detectable antagonism between fludioxonil and propiconazole that would interfere with the mixture's ability to manage sour rot of cherry.

Exceptional Rarity Unveiled: Isolated Fungal Osteomyelitis of Great Toe by Fusarium Solani-Unprecedented Presentation and Exquisite Management Insights

Fungal osteomyelitis is a rare but significant condition, often associated with immunocompromised states or severe systemic illnesses. This condition involves fungal pathogens, primarily Aspergillus, Candida, and Mucor species. Among these, Aspergillus is the most commonly implicated, especially in cases of vertebral osteomyelitis, although fungal infections involving bones like the distal phalanx of the toe are rare. In particular, Fusarium solani, though less frequently reported, has gained recognition for its role in causing localized osteomyelitis in both immunocompetent and immunocompromised patients. A case involving a 57-year-old male with no significant history of immunosuppression is presented. The patient developed localized pain and swelling in the left toe following a minor traumatic injury. Initial radiographic imaging revealed bone erosion, leading to a diagnosis of osteomyelitis. Cultures from bone biopsy identified Fusarium solani as the causative pathogen. This is noteworthy as the fungus, though commonly found in soil, water, and plants, rarely infects bone structures. The patient was treated with a combination of antifungal therapy, including voriconazole and surgical debridement. The rarity of Fusarium solani osteomyelitis and its unusual presentation in an immunocompetent host adds to the complexity of diagnosing fungal infections in osteomyelitis. Treatment typically involves a multidisciplinary approach that combines both medical and surgical interventions. The case highlights the importance of considering fungal pathogens, even in unusual clinical settings, especially when initial treatments fail to provide resolution. Early diagnosis and targeted therapy can improve outcomes, particularly in cases involving uncommon fungal pathogens such as Fusarium solani. This report emphasizes the need for awareness of fungal osteomyelitis in immunocompetent individuals and underscores the critical role of fungal cultures and imaging studies in guiding management.

Development and characterization of nanoemulsions containing Lippia origanoides Kunth essential oil and their antifungal potential against Candida albicans.

Nanoemulsions based on plant essential oils have shown promise as alternatives against fungal pathogens by increasing the solubility and bioavailability of the active compounds of essential oils, which can improve their efficacy and safety. In the present study, we aimed to prepare and characterize nanoemulsions of Lippia origanoides essential oil, and analyze their antifungal activity against C. albicans in planktonic and biofilm form. Additionally, we sought to verify their cytotoxicity. Alginate nanoemulsions were prepared with different concentrations of essential oil, sunflower oil and surfactant to investigate ideal formulations regarding stability and antifungal efficiency. The results showed the nanoemulsions remained stable for longer than 60 days, with acidic pH, particle sizes ranging from 180.17±6.86nm to 497.85±253.50nm, zeta potential from -60.47±2.25 to -43.63±12, and polydispersity index from 0.004 to 0.622. The photomicrographs revealed that the addition of sunflower oil influenced the formation of the particles, forming nanoemulsions. The antifungal results of the essential oil and nanoemulsions showed that the MIC ranged from 0.078 to 0.312mg mL-1. The nanoemulsions were more effective than the free essential oil in eradicating the biofilm, eliminating up to 89.7% of its mass. With regard to cytotoxicity, differences were found between the tests with VERO cells and red blood cells, and the nanoemulsions were less toxic to red blood cells than the free essential oil. These results show that nanoemulsions have antifungal potential against strains of C. albicans in planktonic and biofilm forms.

CRISPR-GRIT: Guide RNAs with Integrated Repair Templates Enable Precise Multiplexed Genome Editing in the Diploid Fungal Pathogen Candida albicans.

Candida albicans, an opportunistic fungal pathogen, causes severe infections in immunocompromised individuals. Limited classes and overuse of current antifungals have led to the rapid emergence of antifungal resistance. Thus, there is an urgent need to understand fungal pathogen genetics to develop new antifungal strategies. Genetic manipulation of C. albicans is encumbered by its diploid chromosomes requiring editing both alleles to elucidate gene function. Although the recent development of CRISPR-Cas systems has facilitated genome editing in C. albicans, large-scale and multiplexed functional genomic studies are still hindered by the necessity of cotransforming repair templates for homozygous knockouts. Here, we present CRISPR-GRIT (Guide RNAs with Integrated Repair Templates), a repair template-integrated guide RNA design for expedited gene knockouts and multiplexed gene editing in C. albicans. We envision that this method can be used for high-throughput library screens and identification of synthetic lethal pairs in both C. albicans and other diploid organisms with strong homologous recombination machinery.

Sphingosine phosphate lyase insufficiency syndrome as a primary immunodeficiency state

Sphingosine phosphate lyase insufficiency syndrome (SPLIS) is a genetic disease associated with renal, endocrine, neurological, skin and immune defects. SPLIS is caused by inactivating mutations in SGPL1, which encodes sphingosine phosphate lyase (SPL). SPL catalyzes the irreversible degradation of the bioactive sphingolipid sphingosine-1-phosphate (S1P), a key regulator of lymphocyte egress. The SPL reaction represents the only exit point of sphingolipid metabolism, and SPL insufficiency causes widespread sphingolipid derangements that could additionally contribute to immunodeficiency. Herein, we review SPLIS, the sphingolipid metabolic pathway, and various roles sphingolipids play in immunity. We then explore SPLIS-related immunodeficiency by analyzing data available in the published literature supplemented by medical record reviews in ten SPLIS children. We found 93% of evaluable SPLIS patients had documented evidence of immunodeficiency. Many of the remainder of cases were unevaluable due to lack of available immunological data. Most commonly, SPLIS patients exhibited lymphopenia and T cell-specific lymphopenia, consistent with the established role of the S1P/S1P1/SPL axis in lymphocyte egress. However, low B and NK cell counts, hypogammaglobulinemia, and opportunistic infections with bacterial, viral and fungal pathogens were observed. Diminished responses to childhood vaccinations were less frequently observed. Screening blood tests quantifying recent thymic emigrants identified some lymphopenic SPLIS patients in the newborn period. Lymphopenia has been reported to improve after cofactor supplementation in some SPLIS patients, indicating upregulation of SPL activity. A variety of treatments including immunoglobulin replacement, prophylactic antimicrobials and special preparation of blood products prior to transfusion have been employed in SPLIS. The diverse immune consequences in SPLIS patients suggest that aberrant S1P signaling may not fully explain the extent of immunodeficiency. Further study will be required to fully elucidate the complex mechanisms underlying SPLIS immunodeficiency and determine the most effective prophylaxis against infection.

Systematic discovery of antibacterial and antifungal bacterial toxins.

Microorganisms use toxins to kill competing microorganisms or eukaryotic cells. Polymorphic toxins are proteins that encode carboxy-terminal toxin domains. Here we developed a computational approach to identify previously undiscovered, conserved toxin domains of polymorphic toxins within 105,438 microbial genomes. We validated nine short toxins, showing that they cause cell death upon heterologous expression in either Escherichia coli or Saccharomyces cerevisiae. Five cognate immunity genes that neutralize the toxins were also discovered. The toxins are encoded by 2.2% of sequenced bacteria. A subset of the toxins exhibited potent antifungal activity against various pathogenic fungi but not against two invertebrate model organisms or macrophages. Experimental validation suggested that these toxins probably target the cell membrane or DNA or inhibit cell division. Further characterization and structural analysis of two toxin-immunity protein complexes confirmed DNase activity. These findings expand our knowledge of microbial toxins involved in inter-microbial competition that may have the potential for clinical and biotechnological applications.

Unveiling the hidden world: How arbuscular mycorrhizal fungi and its regulated core fungi modify the composition and metabolism of soybean rhizosphere microbiome

BackgroundThe symbiosis between arbuscular mycorrhizal fungi (AMF) and plants often stimulates plant growth, increases agricultural yield, reduces costs, thereby providing significant economic benefits. AMF can also benefit plants through affecting the rhizosphere microbial community, but the underlying mechanisms remain unclear. Using Rhizophagus intraradices as a model AMF species, we assessed how AMF influences the bacterial composition and functional diversity through 16 S rRNA gene sequencing and non-targeted metabolomics analysis in the rhizosphere of aluminum-sensitive soybean that were inoculated with pathogenic fungus Nigrospora oryzae and phosphorus-solubilizing fungus Talaromyces verruculosus in an acidic soil.ResultsThe inoculation of R. intraradices, N. oryzae and T. verruculosus didn’t have a significant influence on the levels of soil C, N, and P, or various plant characteristics such as seed weight, crude fat and protein content. However, their inoculation affected the structure, function and nutrient dynamics of the resident bacterial community. The co-inoculation of T. verruculosus and R. intraradices increased the relative abundance of Pseudomonas psychrotolerans, which was capable of N-fixing and was related to cry-for-help theory (plants signal for beneficial microbes when under stress), within the rhizosphere. R. intraradices increased the expression of metabolic pathways associated with the synthesis of unsaturated fatty acids, which was known to enhance plant resistance under adverse environmental conditions. The inoculation of N. oryzae stimulated the stress response inside the soil environment by enriching the polyene macrolide antifungal antibiotic-producing bacterial genus Streptomyces in the root endosphere and upregulating two antibacterial activity metabolic pathways associated with steroid biosynthesis pathways in the rhizosphere. Although inoculation of pathogenic fungus N. oryzae enriched Bradyrhizobium and increased soil urease activity, it had no significant effects on biomass and N content of soybean. Lastly, the host niches exhibited differences in the composition of the bacterial community, with most N-fixing bacteria accumulating in the endosphere and Rhizobium vallis only detected in the endosphere.ConclusionsOur findings demonstrate that intricate interactions between AMF, associated core fungi, and the soybean root-associated ecological niches co-mediate the regulation of soybean growth, the dynamics of rhizosphere soil nutrients, and the composition, function, and metabolisms of the root-associated microbiome in an acidic soil.

Green Medicine: Advancing Antimicrobial Solutions with Diverse Terrestrial and Marine Plant-Derived Compounds

Infectious diseases continue to cause global morbidity and mortality. The rise of drug-resistant pathogens is a major challenge to modern medicine. Plant-based antimicrobials may solve this issue; hence, this review discussed in detail plant-sourced antimicrobial drugs as an alternative toward bacterial, fungal, and viral pathogens. Plant-derived chemicals from various sources such as marine, medicinal, and non-medicinal sources have diverse antimicrobial properties. Complex chemical profiles from these sources allow these molecules to interact with several targets in the microbial pathogens. Due to their multi-component composition, these compounds are more effective and less likely to acquire resistance than single-target antibiotics. Medicinal herbs have long been used for their antimicrobial properties; however, non-medicinal plants have also been identified for their antimicrobial properties. Other interesting new pathways for the identification of antimicrobials include marine plants, which contain a wide variety of metabolites that are both distinct and varied. We have conducted a thorough literature search for the medicinal, non-medicinal, and marine plant-derived molecules with antimicrobial roles from databases which include Scopus, PubMed, Google Scholar, and Web of Science. The review also discussed the synergistic potential of combining these plant-derived compounds with traditional antimicrobial drugs to attenuate the microbial pathogenesis. Based on the existing research and advancements, the review article emphasizes the importance of continuing research into plant-based antimicrobials from these many sources and integrating them with existing therapies to combat the rising threat of drug-resistant infections.

Single-Cell Transcriptomics Unveils Skin Cell Specific Antifungal Immune Responses and IL-1Ra- IL-1R Immune Evasion Strategies of Emerging Fungal Pathogen Candida auris.

Candida auris is an emerging multidrug-resistant fungal pathogen that preferentially colonizes and persists in skin tissue, yet the host immune factors that regulate the skin colonization of C. auris in vivo are unknown. In this study, we employed unbiased single-cell transcriptomics of murine skin infected with C. auris to understand the cell type-specific immune response to C. auris. C. auris skin infection results in the accumulation of immune cells such as neutrophils, inflammatory monocytes, macrophages, dendritic cells, T cells, and NK cells at the site of infection. We identified fibroblasts as a major non-immune cell accumulated in the C. auris infected skin tissue. The comprehensive single-cell profiling revealed the transcriptomic signatures in cytokines, chemokines, host receptors (TLRs, C-type lectin receptors, NOD receptors), antimicrobial peptides, and immune signaling pathways in individual immune and non-immune cells during C. auris skin infection. Our analysis revealed that C. auris infection upregulates the expression of the IL-1RN gene (encoding IL-1R antagonist protein) in different cell types. We found IL-1Ra produced by macrophages during C. auris skin infection decreases the killing activity of neutrophils. Furthermore, C. auris uses a unique cell wall mannan outer layer to evade IL-1R-signaling mediated host defense. Collectively, our single-cell RNA seq profiling identified the transcriptomic signatures in immune and non-immune cells during C. auris skin infection. Our results demonstrate the IL-1Ra and IL-1R-mediated immune evasion mechanisms employed by C. auris to persist in the skin. These results enhance our understanding of host defense and immune evasion mechanisms during C. auris skin infection and identify potential targets for novel antifungal therapeutics.

Candida albicans isolates contain frequent heterozygous structural variants and transposable elements within genes and centromeres.

The human fungal pathogen Candida albicans poses a significant burden on global health, causing high rates of mortality and antifungal drug resistance. C. albicans is a heterozygous diploid organism that reproduces asexually. Structural variants (SVs) are an important source of genomic rearrangement, particularly in species that lack sexual recombination. To comprehensively investigate SVs across clinical isolates of C. albicans, we conducted long read sequencing and genome-wide SV analysis in three distantly related clinical isolates. Our work included a new, comprehensive analysis of transposable element (TE) composition, location and diversity. SVs and TEs are frequently close to coding sequences and many SVs are heterozygous, suggesting that SVs might impact gene and allele-specific expression. Most SVs are uniquely present in only one clinical isolate, indicating that SVs represent a significant source of intra-species genetic variation. We identified multiple, distinct SVs at the centromeres of Chromosome 4 and Chromosome 5, including inversions and transposon polymorphisms. These two chromosomes are often aneuploid in drug resistant clinical isolates, and can form isochromosome structures with breakpoints near the centromere. Further screening of 100 clinical isolates confirmed the widespread presence of centromeric SVs in C. albicans, often appearing in a heterozygous state, indicating that SVs are contributing to centromere evolution in C. albicans Together, these findings highlight that SVs and TEs are common across diverse clinical isolates of C. albicans and that the centromeres of this organism are important sites of genome rearrangement.

Micromanagement: conditions influencing antipathogen function of the skin microbiome in spotted salamanders, Ambystoma maculatum

The emerging fungal pathogen Batrachochytrium salamandrivorans (Bsal) threatens the diversity of amphibians, particularly in North America where it is projected to invade. Amphibian skin defenses include a mucosal layer containing microorganisms that can potentially modulate host response to pathogens such as Bsal. In this study, we focused on the composition of the skin microbiome across life stages of spotted salamanders (Ambystoma maculatum). We also evaluated the stress hormone corticosterone and skin microbiome response to inoculations with Bsal and probiotics at both the larval and juvenile developmental stages, and the response to different environmental conditions. Results indicated that both bacterial and fungal communities found on the skin significantly differed in structure and diversity between life stages of A. maculatum. Exposure to three different probiotics (Bacillus thuringiensis, Chryseobacterium rhizoplanae, and Penicillium sp.) and Bsal evoked shifts in the microbiome of larvae and juveniles, and the metabolite profile of the larval mucosal layer of A. maculatum. Despite changes in the microbiome, all tested probiotics and Bsal were unable to persist on the skin. Larval bacterial microbiomes shifted in response to Bsal and B. thuringiensis with no significant impacts on antifungal function or bacteria richness, however fungi strongly responded to Bsal and B. thuringiensis application. This indicates that developmental shifts in the microbiome can be initiated by microbial applications such as B. thuringiensis, a widely used mosquito larvicide. Overall, experimental results indicate that life stage, growth and development, and environmental conditions appeared to be the main factors driving changes in the amphibian skin microbiome and potential anti-Batrachochytrium function.