Fungal Species Research Articles

Mycoremediation of Acetaminophen by the Fungi of Dothideomycetes sp. Isolate

Resources like air, water, and land are found to be contaminated by a broad variety of pharmaceutical contaminants. The release of industrial effluents, hospital wastewater, and untreated domestic water, which contains medications, colours, pharmaceutical blisters, and heavy metals that are ecotoxic even at concentrations of nanograms to micrograms, is a significant cause of water pollution. Mycoremediation is an operative approach is to detoxify these contaminants. The fungal strain was isolated from pharma industrial wastewater. Identification of the fungi was founded on the morphological and 18s rRNA sequencingtechnique. Isolate shared 99.82 % similarity with Zasmidium cellare and Cladosporium iridis strain CBS 138.40, suggesting the new species of fungi. The strain has received Gen bank accession number of OR081620.1. The fungal strain was found to degrade 85% of acetaminophen (1000 ppm) in 10 days when grown in mineral salt medium with acetaminophen as only carbon source. Glucose was found to enhance sporulation potential but not the degradative potential of the fungi. Identification of biodegradative metabolite was done by using thin layer chromatography, high performance liquid chromatography and liquid chromatography-mass spectroscopy. Mycodegradative metabolite of 4 Aminophenol was identified as acetaminophen.

Preliminary Bioactivity Assessment of Myrothecium Species (Stachybotryaceae) Crude Extracts against Aedes aegypti (Diptera: Culicidae): A First Approach from This Phytopathogenic Fungi.

Mosquitoes, as insect vectors, play a crucial role in transmitting viruses and parasites, leading to millions of human deaths in tropical and subtropical regions worldwide. This study aimed to evaluate the effects of ethanolic extracts of three species within the genus Myrothecium (M. roridum, M. dimerum, and M. nivale) on Aedes aegypti mosquito larvae to assess the inhibitory effect on growth and development, as well as to determine mortality. We quantify the average lethal concentrations and provide a qualitative characterization of the chemical groups responsible for their potential. Phytochemical screening revealed the presence of alkaloids, flavonoids, and terpenoids in the ethanolic extracts of the three fungal species. Tannins were found only in the extracts of M. dimerum and M. roridum. We observed a clear dependence of the effects of the crude extracts on mosquito larvae on the concentrations used and the duration of exposure. The toxic effect was observed after 48 h at a concentration of 800 ppm for both M. dimerum and M. nivale, while M. roridum showed effectiveness after 72 h. All three species within the genus Myrothecium exhibited 100% biological activity after 72 h of exposure at 600 ppm. At lower concentrations, there was moderate growth and development inhibitory activity in the insect life cycle. The study highlights the effectiveness of crude Myrothecium extracts in combating mosquito larvae, with effects becoming apparent between 48 and 72 h of exposure. This initial approach underscores the potential of the fungus's secondary metabolites for further in-depth analysis of their individual effects or synergies between them.

Myco-fabricated silver nanoparticle by novel soil fungi from Saudi Arabian desert and antimicrobial mechanism

Biological agents are getting a noticeable concern as efficient eco-friendly method for nanoparticle fabrication, from which fungi considered promising agents in this field. In the current study, two fungal species (Embellisia spp. and Gymnoascus spp.) were isolated from the desert soil in Saudi Arabia and identified using 18S rRNA gene sequencing then used as bio-mediator for the fabrication of silver nanoparticles (AgNPs). Myco-synthesized AgNPs were characterized using UV–visible spectrometry, transmission electron microscopy, Fourier transform infrared spectroscopy and dynamic light scattering techniques. Their antibacterial activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae were investigated. In atrial to detect their possible antibacterial mechanism, Sodium dodecyl sulfate (SDS-PAGE) and TEM analysis were performed for Klebsiella pneumoniae treated by the myco-synthesized AgNPs. Detected properties of the fabricated materials indicated the ability of both tested fungal strains in successful fabrication of AgNPs having same range of mean size diameters and varied PDI. The efficiency of Embellisia spp. in providing AgNPs with higher antibacterial activity compared to Gymnoascus spp. was reported however, both indicated antibacterial efficacy. Variations in the protein profile of K. pneumoniae after treatments and ultrastructural changes were observed. Current outcomes suggested applying of fungi as direct, simple and sustainable approach in providing efficient AgNPs.

Incidence, Characterization and Pathogenicity of Seed-Borne Fungi of Lentil (<em>Lens culinaris</em> L.) in Pakistan

Lentil (Lens culinaris Medik.) is an important protein-rich Rabi pulse crop after chickpea in Pakistan. Its area and production in the country is decreasing drastically due to its susceptibility to various biotic and abiotic stresses. Among biotic-stresses, seed-borne fungi are the most imperative issue and the accurate identification and characterization of target pathogen isolates/races is crucial for the management of plant diseases in lentils. Therefore, the current study was performed to identify the fungi associated with lentil seeds, to confirm their pathogenicity and to assess their incidence. For this purpose, isolation of fungal pathogens was made on potato dextrose agar medium (PDA) from 56 Lens culinaris L. genotypes. These pathogens were characterized using morphological and molecular techniques and their pathogenicity was performed following blotter paper and agar plate method. Seven fungi belonging to five genera were isolated from lentil seeds (Fusarium avenaceum, Alternaria alternata, Aspergillus flavus, Aspergillus niger, Aspergillus tubingensis, Penicillium citrinum and Bipolaris sorokiniana) of different genotypes. Of these seven fungal species, F. avenaceum, A. tubingensis and B. sorokiniana are new records for lentil seeds from Pakistan. Aspergillus flavus was found to occur in high frequency followed by A. niger, F. avenaceum, P. citrinum, A. alternata, A. tubingensis and B. sorokiniana. Under pathogenicity test on blotter paper and on PDA F. avenaceum, A. flavus, A. niger, A. tubingensis were highly pathogenic, while A. alternata, P. citrinum and B. sorokiniana were found to be less pathogenic. The presence of well-known toxigenic fungal pathogens in lentil seeds suggests the possible risk of contamination of the seeds and enhances the possibility of pre- and post-infections of crop. Therefore, the present study will help to devise effective management strategies to reduce contamination in seeds and also to control the further spread of these pathogens to reduce crop losses.

New species, names, and type designation of rust fungi from New Zealand

ABSTRACT Rust fungi are obligate plant pathogens, usually restricted to a single host species or genus, although some species such as myrtle rust (Austropuccinia psidii) and some cereal and grass rusts (Puccinia spp.) can infect many members of the families Myrtaceae and Poaceae, respectively. The rust life cycle can involve up to five spore stages, both sexual and asexual. The cycle may be completed on a single host (autoecious rust) or require two, unrelated host plants (heteroecious rust) for completion. About 300 rust species are known in New Zealand, with approximately half comprising native rusts (usually pathogenic on native plants) and half being introduced rusts (usually pathogenic on exotic plants). In this paper, 26 new species of rust fungi discovered in New Zealand are described, comprising one species each of Aecidium and Milesina, two of Pucciniastrum, four of Uromyces and 18 of Puccinia. Puccinia ferocis-forsteri is a new heteroecious species producing uredinia and telia on the endemic Carex forsteri and aecia on endemic Urtica ferox. This is only the third native heteroecious rust fungus reported in New Zealand. In addition, two species are transferred to the genus Milesina, three to Uromyces and seven to Puccinia (including a nom. nov.), either following discovery of the sexual (telial) stage of these rusts or considering the most appropriate sexual genus. A new combination is proposed for Coleosporium based on morphology of its urediniospores and molecular data. Epitypes are designated for eight species, lectotypes are selected for five species and a neotype is designated. Apart from one new species on the introduced Fuchsia magellanica all the rusts are considered native to New Zealand. Where appropriate, DNA was extracted and the 28S region (nuclear large subunit ribosomal DNA [nuLSU]) was amplified with the resulting LSU sequences aligned with sequences from GenBank representing most of the major clades.

The influence of car traffic on airborne fungal diversity in Tianjin, China

ABSTRACT Little is known about the effect of car traffic on airborne fungal communities. We investigated the environmental factors affecting the diversity and concentration of airborne fungi at high-traffic density junctions, in Tianjin, China. A total of 244 fungal strains belonging to 78 species and 45 genera of Ascomycota (78.69%) and Basidiomycota (21.31%) were isolated and identified using morphological and molecular analysis. Aspergillus was the species-richest genus, with 9 recorded species, followed by Alternaria and Cladosporium, both with 8 species. Coprinellus radians was the most abundant fungal species, with 31 isolated strains, followed by Alternaria alternata (26 strains), Cladosporium cladosporioides (21), Alternaria compacta (13), and Cladosporium tenuissimum (11). We found a higher diversity and concentration of airborne fungi in the analysed urban air environments when the road traffic was at its highest intensity. Higher level of car traffic resulted in higher concentrations of fungal particles in the air for various taxa, including Alternaria, Aspergillus, and Cladosporium, which are known to cause respiratory allergies and infections. This result suggests that reducing vehicular traffic could be an effective measure to control airborne fungal exposure and microbial pollution.

Soil type and precipitation level have a greater influence on fungal than bacterial diversity in serpentine and non‐serpentine biological soil crusts

AbstractSerpentine soils are characterized by nutrient imbalances and high levels of potentially toxic metals (PTMs). These soils host depauperate plant communities of species with specialized adaptations. Initial studies showed that South African serpentine soils harbor distinct biocrust algal and cyanobacterial species compared to adjacent non‐serpentine soils, with these communities further differing based on high and low precipitation levels. Here, we investigated the bacterial and fungal diversity of biological soil crusts from serpentine and non‐serpentine soils at two precipitation levels. The bacterial and fungal communities were characterized using 16S rDNA and ITS metabarcoding, respectively. No significant differences could be found in bacterial richness and community structure. Nevertheless, bacterial taxa such as Archangium, Candidatus Solibacter, Chthoniobacter, and Microvirga were more abundant in serpentine biocrusts or biocrusts receiving lower precipitation. The fungal community structure was distinct between serpentine and non‐serpentine soils (p = 0.027) and between high and low precipitation (p = 0.018). Furthermore, fungal diversity was lowest in the drier, serpentine biocrusts compared to non‐serpentine (p = 0.001) and serpentine crusts receiving higher precipitation (p = 0.002). The fungal genera, Ramimonilia and Vishniacozyma, which are known to be resistant or tolerant to PTMs and other environmental extremes, were significantly more abundant (p = 0.036 and p = 0.016, respectively) in serpentine biocrusts, with the latter indicating serpentine habitats. This study concluded that soil type influenced the fungal alpha diversity, specifically in the serpentine soil, resulting in a decrease in fungal species richness. Furthermore, precipitation influenced fungal beta diversity by shaping distinct fungal communities found in the biocrusts of serpentine and non‐serpentine soils.

Distinguishing between Wheat Grains Infested by Four Fusarium Species by Measuring with a Low-Cost Electronic Nose.

An electronic device based on the detection of volatile substances was developed in response to the need to distinguish between fungal infestations in food and was applied to wheat grains. The most common pathogens belong to the fungi of the genus Fusarium: F. avenaceum, F. langsethiae, F. poae, and F. sporotrichioides. The electronic nose prototype is a low-cost device based on commercially available TGS series sensors from Figaro Corp. Two types of gas sensors that respond to the perturbation are used to collect signals useful for discriminating between the samples under study. First, an electronic nose detects the transient response of the sensors to a change in operating conditions from clean air to the presence of the gas being measured. A simple gas chamber was used to create a sudden change in gas composition near the sensors. An inexpensive pneumatic system consisting of a pump and a carbon filter was used to supply the system with clean air. It was also used to clean the sensors between measurement cycles. The second function of the electronic nose is to detect the response of the sensor to temperature disturbances of the sensor heater in the presence of the gas to be measured. It has been shown that features extracted from the transient response of the sensor to perturbations by modulating the temperature of the sensor heater resulted in better classification performance than when the machine learning model was built from features extracted from the response of the sensor in the gas adsorption phase. By combining features from both phases of the sensor response, a further improvement in classification performance was achieved. The E-nose enabled the differentiation of F. poae from the other fungal species tested with excellent performance. The overall classification rate using the Support Vector Machine model reached 70 per cent between the four fungal categories tested.

Phenotypic characterization of HAM1, a novel mating regulator of the fungal pathogen Cryptococcus neoformans.

Fungal mating is a vital part of the lifecycle of the pathogenic yeast Cryptococcus neoformans. More than just ensuring the propagation of the species, mating allows for sexual reproduction to occur and generates genetic diversity as well as infectious propagules that can invade mammalian hosts. Despite its importance in the biology of this pathogen, we still do not know all of the major players regulating the mating process and if they are involved or impact its pathogenesis. Here, we identified a novel negative regulator of mating that also affects certain cellular characteristics known to be important for virulence. This gene, which we call HAM1, is widely conserved across the cryptococcal family as well as in many pathogenic fungal species. This study will open new avenues of exploration regarding the function of uncharacterized but conserved genes in a variety of pathogenic fungal species and specifically in serotype A of C. neoformans.

Fungal endo and exochitinase production, characterization, and application for Candida biofilm removal.

Chitinases are promising enzymes for a multitude of applications, including chitooligosaccharide (COS) synthesis for food and pharmaceutical uses and marine waste management. Owing to fungal diversity, fungal chitinases may offer alternatives for chitin degradation and industrial applications. The rapid reproduction cycle, inexpensive growth media, and ease of handling of fungi may also contribute to reducing enzyme production costs. Thus, this study aimed to identify fungal species with chitinolytic potential and optimize chitinase production by submerged culture and enzyme characterization using shrimp chitin. Three fungal species, Coriolopsis byrsina, Trichoderma reesei, and Trichoderma harzianum, were selected for chitinase production. The highest endochitinase production was achieved in C. byrsina after 168h cultivation (0.3 U mL- 1). The optimal temperature for enzyme activity was similar for the three fungal species (up to 45 and 55 ºC for endochitinases and exochitinases, respectively). The effect of pH on activity indicated maximum hydrolysis in acidic pH (4-7). In addition, the crude T. reesei extract showed promising properties for removing Candida albicans biofilms. This study showed the possibility of using shrimp chitin to induce chitinase production and enzymes that can be applied in different industrial sectors.

Diversity and Pathogenicity of Botryosphaeriaceae Species Isolated from Olives in Istria, Croatia, and Evaluation of Varietal Resistance.

During 2021 and 2022, a field investigation was conducted in Istria, Croatia, searching for trees exhibiting signs of Botryosphaeria dieback. Samples of symptomatic trees were collected from 26 different locations and analysed. Isolates that morphologically corresponded to species from the Botryosphaeriaceae family were selected, and detailed morphological characterisation and molecular identification of the isolates were conducted. Based on morphological characteristics and phylogenetic analysis using the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1-α) regions, six species of fungi from the Botryosphaeriaceae family were identified: Botryosphaeria dothidea (Moug. ex Fr.) Ces. & De Not.; Diplodia mutila (Fr.) Fr.; Diplodia seriata De Not.; Dothiorella iberica A.J.L. Phillips, J. Luque & A. Alves; Dothiorella sarmentorum (Fr.) A.J.L. Phillips, Alves & Luque; and Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. This is the first report of D. mutila, Do. sarmentorum, and Do. iberica causing Botryosphaeria dieback on olive trees in Croatia, and the first study investigating the resistance of Croatian olive varieties to species from the Botryosphaeriaceae family. Pathogenicity testing of selected isolates and assessment of variety resistance were conducted on four different olive varieties, namely Buža, Istarska bjelica, Leccino, and Rosinjola, using representative isolates of the mentioned species. The most aggressive species was found to be N. parvum. Olive varieties exhibited differences in susceptibility depending on the fungus they were infected with.

Fungal cryopreservation across 61 genera: Practical application and method evaluation

ABSTRACT Fungi occupy important environmental, cultural, and socioeconomic roles. However, biological research of this diverse kingdom has lagged behind that of other phylogenetic groups. This is partially the result of the notorious difficulty in culturing a diverse array of filamentous fungal species due to their (i) often unpredictable growth, (ii) unknown preferences for culturing conditions, and (iii) long incubation times compared with other microorganisms such as bacteria and yeasts. Given the complexity associated with concurrently culturing diverse fungal species, developing practical methods for preserving as many species as possible for future research is vital. The widely accepted best practice for preserving fungal tissue is the use of cryogenic biobanking at −165 C, allowing for the preservation and documentation of stable genetic lineages, thus enabling long-term diversity-centered research. Despite the extensive literature on fungal cryopreservation, substantial barriers remain for implementation of cryogenic biobanks in smaller mycological laboratories. In this work, we present practical considerations for the establishment of a fungal culture biobank, as well as provide evidence for the viability of 61 fungal genera in cryogenic storage. By providing a pragmatic methodology for cryogenically preserving and managing many filamentous fungi, we show that creating a biobank can be economical for independently owned and operated mycology laboratories, which can serve as a long-term resource for biodiversity, conservation, and strain maintenance.

Crecimiento micelial y producción de lacasa y peroxidasas por Pleurotus ostreatus y Agrocybe aegerita en fermentación líquida

Mycelial growth of Pleurotus ostreatus and Agrocybe aegerita in malt extract broth was evaluated. Laccase, manganese peroxidase (MnP), lignin peroxidase (LiP) and unspecific peroxygenase (UnP) activities were determined in both strains, using biochemical and polyacrylamide gel electrophoresis techniques. No strain showed MnP, LiP, and UnP activities. P. ostreatus had higher biomass production and laccase yield values than those shown by A. aegerita. P. ostreatus produced approximately twice as much biomass (Xmax) compared to A. aegerita (9.12 and 5.27 g/L, respectively). Maximum laccase activity (Emax) was nine-fold higher in P. ostreatus than in A. aegerita (106.5 and 12.23 U/L, respectively). P. ostreatus and A. aegerita showed 3 bands (estimated to be approximately 42, 55, 85 and 212 kDa) and one band (estimated to be approximately 35.2 kDa) with laccase activity, respectively. These results show that laccases are constitutively produced in both fungal species, whereas MnP, LiP and UnP are inducible enzymes. P. ostreatus has a great potential in the biotechnological production of laccases.

Novel powdery mildew of cotton (Gossypium hirsutum) caused by Phyllactinia gossypina sp. nov. in Brazil.

Cotton (Gossypium hirsutum, Malvaceae) is the most important fiber crop in the world. There are published records of many fungal pathogens attacking Gossypium spp., causing numerous diseases, including powdery mildews. Recently, in 2022, non-cultivated spontaneous G. hirsutum plants bearing powdery mildews symptoms were found at roadsides in two municipalities of the state of Minas Gerais (Brazil): Varginha and Ubá. Such localities are situated ca. 260km apart, suggesting a broader distribution of this fungus-host association in Brazil. Samples were taken to the laboratory, and an Ovulariopsis-like, asexual stage of Phyllactinia, was identified forming amphigenous colonies, that were more evident, white and cottony, abaxially. Morphological and molecular data- of the ITS and LSU regions- have shown that colonies from those two samples were of the same fungus species, belonging to a previously unknown species of Erysiphaceae (Ascomycota). The fungus fits into the Phyllactinia clade and is described herein as the new species Phyllactinia gossypina sp. nov. This new species belongs to the 'basal Phyllactinia group', a lineage that includes species known only from the Americas. This report expands the list of pathogenic fungi on cotton. It is early to anticipate whether this new powdery mildew represents a threat to cultivated cotton, which is a major crop in Brazil. Nevertheless, further studies about its infectivity to commercial cotton varieties are recommended, since all known Erysiphaceae are specialized obligate plant parasites and several species cause major losses to important crops.

Assembly and comparative analyses of the Geosiphon pyriformis metagenome.

Geosiphon pyriformis, a representative of the fungal sub-phylum Glomeromycotina, is unique in its endosymbiosis with cyanobacteria within a fungal cell. This symbiotic relationship occurs in bladders containing nuclei of G. pyriformis, Mollicutes-like bacterial endosymbionts (MRE), and photosynthetically active and dividing cells of Nostoc punctiforme. Recent genome analyses have shed light on the biology of G. pyriformis, but the genome content and biology of its endosymbionts remain unexplored. To fill this gap, we gathered and examined metagenomic data from the bladders of G. pyriformis, where N. punctiforme and MRE are located. This ensures that our analyses are focused on the organs directly involved in the symbiosis. By comparing this data with the genetic information of related cyanobacteria and MREs from other species of Arbuscular Mycorrhizal Fungi, we aimed to reveal the genetic content of these organisms and understand how they interact at a genetic level to establish a symbiotic relationship. Our analyses uncovered significant gene expansions in the Nostoc endosymbiont, particularly in mobile elements and genes potentially involved in xenobiotic degradation. We also confirmed that the MRE of Glomeromycotina are monophyletic and possess a highly streamlined genome. These genomes show dramatic differences in both structure and content, including the presence of enzymes involved in environmental sensing and stress response.

Inventory of Macroscopic Fungi in Protected Forest Areas Malaka District Chartery

Mushrooms are eukaryotic organisms with spores, without chlorophyll that reproduce sexually and asexually, mushrooms can be used by humans as food ingredients and ingredients for traditional and modern medicine. The purpose of this study was to determine the type and habitat characteristic of macroscopic fungi conducted in the kateri procted forest area Malaka Regency. Using exploration and identification methods where the plant identification process is a process of matching a plant according to a particular taxonomi on each research transect line. There are 28 species of macroscopic fungi namely Dacrymyces stilatus, Auricularia auricula-judae, Xylaria hypoxylon, Daldinia concentrica, Microporus xanthopus, Earliella scabrosa, Pycnoporus sanguineus, Polyporus alveolaris, Lentinus squarrosulus, Hexagonia tenuis, Trametes pubescens, Tyromyces chioneus, Marasmius oreades, Marasmius haematocephalus, Baespora myosura, Marasmius rotula, Marasmius andorsaceus, Trogia infundibiliformis, Leucocoprinus brebissonii, Lepiota helveola. Agaricus micromegathus, Bolbitius vitelinus, Mycena albicocapilaris, Crepidotus variabilis, Parasola plicatilis, Coprinopsis fragilisimus, Lycoperdon pyriforme, Pleurotus djamor. Divided into 5 Orders namely Dacrymycetales, Auriculariales, Xylariales, Polyporales, Agaricales. And 12 family namely Dacrymycetaceae, Auriculariaceae, Xylariaceae, Polyporaceae, Marasmiaceae, Agaricaceae, Bolbitiaceae, Mycenaceae, Crepidotaceae, Psathyrellaceae, Schizophyllaceae, Pleurotaceae. The most abundant habitat Characteristics were on rotten wood branches as many 9 species, on dead wood stem many as 8 species, in leaf little many as 4 species and in soil with hums many as 7 species.

Biodegradation of Difenoconazole Using Fungal-Bacterial Consortia

Abstract Pesticides are used all over the world in agricultural operations to reduce the damage of pests and to improve the productivity of agricultural crops. The excessive use of these chemical compounds has led to the deterioration of soil and increase the level of pollution of water and air resources. Moreover, the continuous use and non-target toxicity of pesticides has become a major concern to the agricultural ecosystem that directly hinders the productivity of agricultural products. Therefore, this study aimed to isolate and identify fungal and bacterial species with the ability to biodegrade fungicide via conducting preliminary tests to find out which of them have the ability to grow in a treated medium with a fungicide Difenoconazole, which belongs to the group of the pesticides Triazoles. The fungi and bacteria that succeeded in growing were as follow Aspergillus flavus, Aspergillus ochraceus, Bacillus cereus S1 and Bacillus cereus S2 were identified by extracting their genomic DNA and applying the primers ITS and 16s rRNA for both examined fungi and bacteria, respectively. The fungi were laboratory adapted to multiple concentrations reaching the highest concentration of 550 mgL-1, while bacteria reached 1200 mgL-1 laboratory to break down the pesticide. The results of the combinations tested as the B. cereus S1 and B. cereus S2 were the fastest growing in the liquid medium PDB at the concentration of 325 mgL-1. While the combination A. flavus and B. cereus S1 had the highest biodegrading ability at the same concentration, which led to the disappearance of the pesticide Difenoconazole turbidity in the liquid medium as a result of its consumption by fungal and bacterial isolates. As for the treatment of the pesticide in the solid PDA medium to know its effect on the growth of fungi and the formation of spores, the results revealed that fungal isolates increase their vegetative growth when the concentration increases, the production of fungal spores decreases, and indicates the ability of the fungi to use the pesticide as an energy source.

Environmental Fungal Contamination Characterization of Three Inpatient Units Utilizing Optimized Detection Methods

Background: Environmental sampling and detection methods for fungi in healthcare settings are not well-established. We previously refined methods for fungal sampling and detection in a controlled laboratory environment and aimed to validate them in a real-world healthcare setting. Methods: We performed a microbiological analysis of air and surfaces in three inpatient units at a tertiary care center. Surface samples were obtained with foam sponges from 3 locations in patient rooms (Patient bedrails, bathroom floor, HVAC export) and 5 locations in units (HVAC exports 3x, clean linen storage, soiled linen storage). Air samples were taken with an active air sampler directly below HVAC exports. Sponges were processed using the stomacher technique. Samples underwent DNA extraction followed by qPCR with FungiQuant primers targeting the 18S rRNA gene. Amplicons from positive samples were sequenced (NextSeq 1000, 300bp PE) and SmartGene databases were used to interpret sequence data. For comparison to culture methods, samples were also plated onto Sabouraud and HardyCHROM Candida + auris medias. Fungal growth underwent DNA extraction, 18S PCR and Sanger sequencing for genus and species identification. Results: A total of 85 samples were obtained, from 15 patient rooms and three units resulting in 61 surface and 24 air samples. Patients in study rooms had a median age of 53, 9 (60%) were male, and no patients had an invasive fungal infection during their hospital encounter. 44 (53%) and 39 (46%) samples were positive for fungi via qPCR and culture, respectively. Of the 44 positive qPCR samples, microbiome analyses identified at least one fungi to the species, genus and family levels in 43 (98%), 28 (64%), 18 (41%) samples, respectively (Table 1). 114 total isolates were identified of which the most common were Mallassezia restricta (30 [26%]), Malassezia globose (29 [25%]), and Pennicillium paradoxum (4 [4%]). 39 genera were identified of which the most common were Mucor (19 [49%]) and Candida (8 [21%]). Of the 39 culture positive samples, 90 total isolates were recovered. The most common species were Paradendryphiella arenariae (19 [21%]), Aspergillus niger (12 [13%]) and Penicillium commune (12 [13%]). Conclusion: These results demonstrate the presence of diverse fungal species in both air and surface samples across inpatient units. Higher sensitivity was noted utilizing qPCR, however, identified genera and species were markedly different between qPCR and culture methods. Larger studies are needed to assess the efficacy of qPCR for fungal detection in the healthcare environment.

Multikingdom characterization of gut microbiota in patients with rheumatoid arthritis and rheumatoid arthritis-associated interstitial lung disease.

Rheumatoid arthritis-associated interstitial lung disease (RA-ILD) is a serious and common extra-articular disease manifestation. Patients with RA-ILD experience reduced bacterial diversity and gut bacteriome alterations. However, the gut mycobiome and virome in these patients have been largely neglected. In this study, we performed whole-metagenome shotgun sequencing on fecal samples from 30 patients with RA-ILD, and 30 with RA-non-ILD, and 40 matched healthy controls. The gut bacteriome and mycobiome were explored using a reference-based approach, while the gut virome was profiled based on a nonredundant viral operational taxonomic unit (vOTU) catalog. The results revealed significant alterations in the gut microbiomes of both RA-ILD and RA-non-ILD groups compared with healthy controls. These alterations encompassed changes in the relative abundances of 351 bacterial species, 65 fungal species, and 4,367 vOTUs. Bacteria such as Bifidobacterium longum, Dorea formicigenerans, and Collinsella aerofaciens were enriched in both patient groups. Ruminococcus gnavus (RA-ILD), Gemmiger formicilis, and Ruminococcus bromii (RA-non-ILD) were uniquely enriched. Conversely, Faecalibacterium prausnitzii, Bacteroides spp., and Roseburia inulinivorans showed depletion in both patient groups. Mycobiome analysis revealed depletion of certain fungi, including Saccharomyces cerevisiae and Candida albicans, in patients with RA compared with healthy subjects. Notably, gut virome alterations were characterized by an increase in Siphoviridae and a decrease in Myoviridae, Microviridae, and Autographiviridae in both patient groups. Hence, multikingdom gut microbial signatures showed promise as diagnostic indicators for both RA-ILD and RA-non-ILD. Overall, this study provides comprehensive insights into the fecal virome, bacteriome, and mycobiome landscapes of RA-ILD and RA-non-ILD gut microbiota, thereby offering potential biomarkers for further mechanistic and clinical research.

Evaluation of antifungal activity of plant extracts against plant pathogen associated with sweet orange (Citrus sinensis L.), in Jimma, Western Ethiopia

BackgroundSweet oranges are a vital agricultural commodity, but they are susceptible to various pathogenic fungi that can cause significant economic losses. This study aimed to evaluate the antifungal activity of plant extracts against pathogenic fungi attacking sweet oranges in Jimma, Ethiopia.Materials and methodsVarious plant extracts were obtained from locally available plant species and tested against selected pathogenic fungi using standard in vitro antifungal assays. The results revealed varying degrees of antifungal activity among the plant extracts, with some showing promising inhibitory effects against the targeted fungi. Statistical analysis indicated significant differences in antifungal efficacy among the tested extracts.ResultsThe study found that Penicilium spp., Aspergillus spp., Rhizopus spp., Mucor spp., and Fusarium spp, were the predominant fungal species. The study found that the extracts of Vernonia amygdalina, Ginger oficealae, and Pterolobium stalatam were effective in inhibiting mycelial growth against these pathogens. This suggests the use of these medicinal plants for the preservation and control of sweet orange fruit in most of the Ethiopian region.ConlusionThese findings suggest the potential of certain plant extracts as natural alternatives to synthetic fungicides for managing fungal diseases in sweet orange cultivation. Further research is needed to elucidate the mechanisms of action of these plant extracts and optimize their application in agricultural practices. This study contributes to the development of sustainable and eco-friendly strategies for plant disease management in the region.