Tristan W. Wang

Candida auris is associated with life-threatening invasive disease due to high level of drug resistance. We present a clinical case of C. auris multi-drug resistance development in a single patient acquired during antifungal treatment. Five isolates were prospectively recovered from a transplant patient receiving antifungal therapy over a one-year period. While isolates were initially only resistant to fluconazole, the terminal isolate became resistant to caspofungin and amphotericin B with significant increase in micafungin MIC. Sequencing of ERG11 and FKS1 genes identified mutations associated with fluconazole and echinocandin resistance in the multi-drug-resistant isolate, underscoring the threat of therapy-induced development of resistance.

Candida auris is an emerging nosocomial fungal pathogen associated with life-threatening invasive disease due to its persistent colonization, high level of transmissibility and multi-drug resistance. Aggregative and non-aggregative growth phenotypes for C. auris strains with different biofilm forming abilities, drug susceptibilities and virulence characteristics have been described. Using comprehensive transcriptional analysis we identified key cell surface adhesins that were highly upregulated in the aggregative phenotype during in vitro and in vivo grown biofilms using a mouse model of catheter infection. Phenotypic and functional evaluations of generated null mutants demonstrated crucial roles for the adhesins Als4112 and Scf1 in mediating cell-cell adherence, coaggregation and biofilm formation. While individual mutants were largely non-aggregative, in combination cells were able to co-adhere and aggregate, as directly demonstrated by measuring cell adhesion forces using single-cell atomic force spectroscopy. This co-adherence indicates their role as complementary adhesins, which despite their limited similarity, may function redundantly to promote cell-cell interaction and biofilm formation. Functional diversity of cell wall proteins may be a form of regulation that provides the aggregative phenotype of C. auris with flexibility and rapid adaptation to the environment, potentially impacting persistence and virulence.

The oral cavity remains an underappreciated site for SARS-CoV-2 infection despite the myriad of oral conditions in COVID-19 patients. Recently, SARS-CoV-2 was shown to replicate in the salivary gland cells causing tissue inflammation. Given the established association between inflammation and microbiome disruption, we comparatively profiled oral microbial differences at a metagenomic level in a cohort of hospitalized COVID-19 patients and matched healthy controls. Specifically, we aimed to evaluate colonization by the opportunistic fungal pathogen Candida albicans, the etiologic agent of oral candidiasis. Comprehensive shotgun metagenomic analysis indicated that, overall, COVID-19 patients exhibited significantly reduced bacterial and viral diversity/richness; we identified 12 differentially abundant bacterial species to be negatively associated with COVID-19, and the functional pathways of certain bacteria to be highly associated with COVID-19 status. Strikingly, C. albicans was recovered from approximately half of the COVID-19 subjects but not from any of the healthy controls. The prevalence of Candida is likely linked to immune hypo-dysregulation caused by COVID-19 favoring Candida proliferation, warranting investigations into the interplay between Candida and SARS-CoV2 and potential therapeutic approaches directed toward oral candidiasis. Collectively, our findings prompt a reassessment of oral opportunistic infection risks during COVID-19 disease and their potential long-term impacts on oral health.

Candida auris is an emerging nosocomial fungal pathogen associated with life-threatening invasive disease due to its persistent colonization, high level of transmissibility and multi-drug resistance. Aggregative and non-aggregative growth phenotypes for C. auris strains with different biofilm forming abilities, drug susceptibilities and virulence characteristics have been described. Using comprehensive transcriptional analysis we identified key cell surface adhesins that were highly upregulated in the aggregative phenotype during in vitro and in vivo grown biofilms using a mouse model of catheter infection. Phenotypic and functional evaluations of generated null mutants demonstrated crucial roles for the adhesins Als5 and Scf1 in mediating cell-cell adherence, coaggregation and biofilm formation. While individual mutants were largely non-aggregative, in combination cells were able to co-adhere and aggregate, as directly demonstrated by measuring cell adhesion forces using single-cell atomic force spectroscopy. This co-adherence indicates their role as complementary adhesins, which despite their limited similarity, may function redundantly to promote cell-cell interaction and biofilm formation. Functional diversity of cell wall proteins may be a form of regulation that provides the aggregative phenotype of C. auris with flexibility and rapid adaptation to the environment, potentially impacting persistence and virulence.

The postharvest fruit pathogen Paecilomyces niveus produces ascospores that can survive some pasteurization temperatures, spoil fruit products, and contaminate them with patulin, an FDA-regulated mycotoxin. Preventing P. niveus from entering food systems requires a robust detection method to effectively determine sources of P. niveus spoilage and disease inoculum. We designed a new robust and culture-independent detection method using species-specific primers (PnPATf/r) based on the patK gene, encoding a 6-methylsalicylic acid synthase, in P. niveus , for use in a rapid qPCR assay. Primer specificity was validated using 24 different P. niveus isolates and 16 other important food spoilage and fruit pathogenic fungi. The threshold for detection of qPCR was 18 genome equivalents. To further validate our new detection method, we demonstrate its use in detecting P. niveus in infected fruits, infested soils and ciders, and in fruit arising from apple blossoms sprayed with a P. niveus spore suspension. Results from this study may help fruit producers address spoilage and patulin contamination by this food spoiling fungus. Highlights: New primers specific to Paecilomyces niveus (PnPATf/r) were developed based on the patK gene A qPCR assay to detect P. niveus was validated, and shown to be able to detect quantities of P. niveus DNA as low as 18 genome equivalents The new qPCR assay was used to investigate the ability of P. niveus ascospores to infect strawberry fruits and enter apple fruits through apple blossom infestation

Hard apple cider is considered to be a low-risk product for food spoilage and mycotoxin contamination due to its alcoholic nature and associated food sanitation measures. However, the thermotolerant mycotoxin-producing fungus Paecilomyces niveus may pose a significant threat to hard cider producers. Pa. niveus is known to infect apples ( Malus x domestica ), and previous research indicates that it can survive thermal processing and contaminate finished apple juice with the mycotoxin patulin. To determine if hard apple cider is susceptible to a similar spoilage phenomenon, cider apples were infected with Pa. niveus or one of three patulin-producing Penicillium species and the infected fruits underwent benchtop fermentation. Cider was made with lab inoculated Dabinett and Medaille d’Or apple cultivars, and patulin was quantified before and after fermentation. Results show that all four fungi can infect cider apples and produce patulin, some of which is lost during fermentation. Only Pa. niveus was able to actively grow throughout the fermentation process. To determine if apple cider can be treated to hinder Pa. niveus growth, selected industry-grade sanitation measures were tested, including chemical preservatives and pasteurization. High concentrations of preservatives inhibited Pa. niveus growth, but apple cider flash pasteurization was not found to significantly impact spore germination. This study confirms that hard apple cider is susceptible to fungal-mediated spoilage and patulin contamination. Pa. niveus should be of great concern to hard apple cider producers due to its demonstrated thermotolerance, survival in fermentative environments, and resistance to sanitation measures. Highlights: Apple fruits of traditional cider cultivars Dabinett and Medaille d’Or were found to be susceptible to infection by three patulin-producing Penicillium spp. and Paecilomyces niveus Pa. niveus can grow in finished fermented hard cider at 5.22% ethanol Patulin levels in cider were reduced by fermentation but still exceeded 50 µg/kg, a maximum limit set by various regulatory agencies Pa. niveus was observed to be able to grow in low concentrations of three preservatives: potassium sorbate, sulfur dioxide, and sodium benzoate

Paecilomyces rot of apples is a postharvest disease caused by Paecilomyces niveus, a problematic spoiling agent of fruit juices and derivatives. Processing fruits infected with Paecilomyces rot can lead to juices contaminated with P. niveus ascospores. These ascospores are heat resistant and may survive food processing and germinate in finished products. Because the fungus produces the mycotoxin patulin, juice spoilage by P. niveus is an important health hazard. Little is known about the disease biology and control mechanisms of this recently described postharvest disease. The range of fruit products contaminated by P. niveus and patulin led us to hypothesize that the host range of Paecilomyces rot is broader than previously thought. Following Koch’s postulates, we determined that multiple untested rosaceous fruits and popular apple cultivars are susceptible to Paecilomyces rot infection and that these infected fruits contain significant levels of patulin. We also observed that two closely related food spoiling fungi, Paecilomyces fulvus and Paecilomyces variotti, were unable to infect, cause symptoms in, or grow in wounded fruits. Therefore, we challenge the assumption that P. niveus spoilage inoculum is introduced to foods solely through environmental sources, and we show that other economically important rosaceous fruits, peaches, pears, sweet cherries, and sour cherries, are susceptible to infection and can also be sources of spoilage inoculum. Our results highlight the unique abilities of P. niveus to infect a variety of fruits, produce patulin, and form resistant spores capable of spoiling normally shelf-stable products.

Desiccation tolerance was a key trait that allowed plants to colonize land. However, little is known about the transition from desiccation tolerant non-vascular plants to desiccation sensitive vascular ones. Filmy ferns (Hymenophyllaceae) represent a useful system to investigate how water-stress strategies differ between non-vascular and vascular stages within a single organism because they have vascularized sporophytes and nonvascular gametophytes that are each capable of varying degrees of desiccation tolerance. To explore this, we surveyed sporophytes and gametophytes of 19 species (22 taxa including varieties) of filmy ferns on Moorea (French Polynesia) and used chlorophyll fluorescence to measure desiccation tolerance and light responses. We conducted phylogenetically informed analyses to identify differences in physiology between life stages and growth habits. Gametophytes had similar or less desiccation tolerance (ability to recover from 2 days desiccation at − 86 MPa) and lower photosynthetic optima (maximum electron transport rate of photosystem II and light level at 95% of that rate) than sporophytes. Epiphytes were more tolerant of desiccation than terrestrial species in both life stages. Despite their lack of greater physiological tolerances, gametophytes of several species occurred over a wider elevational range than conspecific sporophytes. Our results demonstrate that filmy fern gametophytes and sporophytes differ in their physiology and niche requirements, and point to the importance of microhabitat in shaping the evolution of water-use strategies in vascular plants.

Paecilomyces rot of apples is a postharvest disease caused by Paecilomyces niveus , a problematic spoiling agent of fruit juices and derivatives. The fungus produces ascospores that can survive food processing and germinate in finished fruit products. Processing apple fruits infected with Paecilomyces rot can lead to P. niveus contaminated juices. Because the fungus produces the mycotoxin patulin, juice spoilage by P. niveus is an important health hazard. Little is known about the disease biology and control mechanisms of this recently described postharvest disease. Following Koch’s postulates, we determined that a range of previously untested rosaceous fruits and popular apple cultivars are susceptible to Paecilomyces rot infection. We also observed that two closely related food spoiling fungi, Paecilomyces fulvus and Paecilomyces variotti , were unable to infect, cause symptoms in, or reproduce in wounded fruits. Our results highlight the unique abilities of Paecilomyces niveus to infect a variety of fruits, produce patulin, and form highly-resistant spores capable of spoiling normally shelf-stable products.