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Invasive aspergillosis is a threat for immunocompromised patients. We present a case series of aggressive cerebral vasculitis caused by Aspergillus spp. infection in immunocompromised patients.We present a retrospective case series of three autopsy-proven invasive cerebral aspergillosis with diffuse vasculitis affecting large caliber cerebral vessels.Three patients were immunosuppressed: one on rituximab, one on corticosteroids, and one with a renal transplant. Two of these patients were diagnosed with cerebral aspergillosis on postmortem.Aspergillus cerebral vasculitis is a rare form of invasive aspergillosis that should be considered in an immunocompromised individual with suggestive lesions on imaging. It should be suspected as a possible cause of aseptic neutrophil meningitis.

Ochratoxin A (OTA), primarily produced by the fungi belonging to the species of Aspergillus and Penicillium, is one of the most common mycotoxins found in cereals and fruits. In addition to resulting in huge economic losses, OTA contamination also poses considerable threat to human and livestock health. Microbial degradation of mycotoxins has been considered with great potential in mycotoxins decontamination. In a previous study, Bacillus velezensis E2 was isolated by our laboratory and showed appreciable inhibitory effect on Aspergillus flavus growth and aflatoxin production in rice grains. In this study, B. velezensis E2 was investigated for its ability to remove OTA and biocontrol against the ochratoxigenic Aspergillus westerdijkiae fc-1. The results revealed that B. velezensis E2 has considerable inhibitory effect on A. westerdijkiae fc-1 both on PDA medium and pear fruits, with inhibitory rate of 51.7% and 73.9%, respectively. In addition, its ability to remove OTA was evaluated in liquid medium and the results showed that more than 96.1% of OTA with an initial concentration of 2.5 μg/mL could be removed by B. velezensis E2 in 48 h. Further experiments revealed that enzymatic transformation and alkaline hydrolysis might be the main mechanisms related to OTA degradation by B. velezensis E2, with ring open ochratoxin α (OP-OTα) as a possible degradation product. Our study indicated that the B. velezensis E2 strain could be a potential bacterial candidate in biodegradation of OTA and biocontrol against A. westerdijkiae fc-1.

CRISPR/Cas technology is a versatile tool for genome engineering in many organisms, including filamentous fungi. Cpf1 is a multi-domain protein of class 2 (type V) RNA-guided CRISPR/Cas endonuclease, and is an alternative platform with distinct features when compared to Cas9. However, application of this technology in filamentous fungi is limited. Here, we present a single CRISPR/Cpf1 plasmid system in Aspergillus aculeatus strain TBRC 277, an industrially relevant cell factory. We first evaluated the functionality of three Cpf1 orthologs from Acidaminococcus sp. BV3L6 (AsCpf1), Francisella tularensis subsp. novicida U112 (FnCpf1), and Lachnospiraceae bacterium (LbCpf1), in RNA-guided site-specific DNA cleavage at the pksP locus. FnCpf1 showed the highest editing efficiency (93 %) among the three Cpf1s. It was further investigated for its ability to delete a 1.7 kb and a 0.5 kb from pksP and pyrG genes, respectively, using two protospacers targeting these gene loci in a single crRNA array. Lastly, simultaneous editing of three sites within TBRC 277 genome was performed using three guide sequences targeting these two genes as well as an additional gene, kusA, which resulted in combined editing efficiency of 40 %. The editing of the NHEJ pathway by targeting kusA to generate a NHEJ-deficient strain of A. aculeatus TBRC 277 improved gene targeting efficiency and yielded more precise gene-editing than that of using wild-type strain. This promising genome-editing system can be used for strain improvement in industrial applications such as production of valuable bioproducts.

The discovery of itaconic acid as a product of citric acid pyrolytic distillation in 1837 opened the possibility of using it as a polymer building block. Itaconic acid, featuring two carboxylic acids and an unsaturated group, can potentially be used as a building block in several chemical syntheses, with a particular emphasis on polymer manufacture. The elucidation of biochemical pathways originating from itaconic acid, first in Aspergillus terreus and, recently, in several species of the Ustilago genus, has intensified and diversified research focused on microbe-based itaconic acid production, including at an industrial scale. These efforts include the engineering of naturally producing species/strains along with the exploration of other species that do not naturally produce itaconic acid but may offer potential benefits. The use of renewable wastes or sugar-enriched residues as substrates to produce itaconic acid, from a circular bioeconomy perspective, is another important aspect of the advancements in microbial itaconic acid production. This review provides an overview of the achievements as well as the challenges concerning the engineering of the producing strains/species, substrate selection, optimisation of bioreactor operation, and downstream itaconic acid purification methods. info:eu-repo/semantics/publishedVersion

Fungal corneal ulcers are an uncommon, yet challenging, cause of vision loss. In the United States, geographic location appears to dictate not only the incidence of fungal ulcers, but also the fungal genera most encountered. These patterns of infection can be linked to environmental factors and individual characteristics of fungal organisms. Successful management of fungal ulcers is dependent on an early diagnosis. New diagnostic modalities like confocal microscopy and polymerase chain reaction (PCR) are being increasingly used to detect and identify infectious organisms. Several novel therapies, including crosslinking and light therapy, are currently being tested as alternatives to conventional antifungal medications. We explore the biology of Candida, Fusarium, and Aspergillus, the three most common genera of fungi causing corneal ulcers in the United States and discuss current treatment regimens for the management of fungal keratitis.

The airway epithelium maintains tight barrier integrity to prevent penetration of pathogens; thus, impairment of the barrier function is an important and common histological feature in asthmatic patients. Proteolytic allergens from fungi, pollen, and house dust mites can disrupt epithelial barrier integrity, but the mechanism remains unclear. Aspergillus oryzae protease (AP)-induced mitochondrial reactive oxygen species (ROS) contribute to the epithelial inflammatory response. However, as mitochondrial ROS affect various cellular functions, such as metabolism, cell death, cell proliferation, and redox homeostasis through signal transduction, it is difficult to understand the detailed action mechanism of AP by measuring changes in a single gene or protein of a specific signaling pathway. Moreover, mitochondrial ROS can directly oxidize DNA to activate transcription, thereby affecting the expression of various genes at the transcriptional level. Therefore, we conducted whole-genome analysis and used a network-based approach to understand the effect of AP and AP-induced mitochondrial ROS in human primary airway epithelial cells and to evaluate the mechanistic basis for AP-mediated epithelial barrier dysfunction. Our results indicate that production of mitochondrial ROS following AP exposure induce mitochondrial dysfunction at an early stage. Over time, changes in genome expression were further expanded without remaining mitochondrial ROS. Specifically, genes involved in the apoptotic functions and intercellular junctions were affected, consequently impairing the cellular barrier integrity. This change was recovered by scavenging mitochondrial ROS at an early point after exposure to AP. In conclusion, our findings indicate that instantly increased mitochondrial ROS at the time of exposure to allergenic proteases consequently induces epithelial barrier dysfunction at a later time point, resulting in pathological changes. These data suggest that antioxidant therapy administered immediately after exposure to proteolytic antigens may be effective in maintaining epithelial barrier function.

Background Indoor allergens are important components of airborne allergens, and studies have linked allergic respiratory diseases (ARDs) and exposure to aeroallergens. Objectives This study sought to identify fungi in house dust from four regions in Baghdad city, and to assess the risks of ARD and asthma associated with indoor fungi. Methods House dust was collected from 84 homes located in four major areas in Baghdad; locations 1, 2, 3 and 4 (north, west, south-west and south of Baghdad, respectively). The dust samples were cultured to identify fungal genera. Results Seventy-nine mold isolates belong to nine genera (Aspergillus, Rhizopus, Penicillium, Chrysonilia, Mucor, Fusarium, Cladosporium, Alternaria and Helminthosporium), as well as five yeast isolates, were identified. Aspergillus was the most common type of mold (56.0%), while Alternaria and Helminthosporium were the least frequent (each with 1%). The distribution of these fungi showed variation according to location, but the difference was not significant. ARD in general and asthma in particular were detected in 50.0 and 14.3% of households, respectively. Both types of allergy were significantly associated with mold genera and with the number of households and rooms, as well as the availability of gardens and trees. Conclusions The study indicated that fungi are significant indoor determinants of ARD and asthma in the city of Baghdad.

Primary cutaneous aspergillosis is an uncommon opportunistic infection, generally seen in immunocompromised individuals. However, many atypical presentations have been reported recently particularly with the increased domain of immunosuppression. Sporotrichoid pattern of aspergillosis where it invades deep lymphatics have rarely been reported and never in immunocompetent individuals previously. We hereby report a case of a 29 years old immunocompetent individual with no comorbidities who presented with multiple painful erythematous papules and nodules over the left upper limb in a sporotrichoid pattern. Microscopy revealed branched septate hyphae and culture on Sabouraud dextrose agar grew powdery greenish colonies which showed hyaline branched septate hyphae with brush-like conidiophores and globose conidia in chains. Histopathology from one of the nodules was consistent with deep fungal infection. A diagnosis of Aspergillus chivalieri was made based on 18S rRNA sequencing of the isolate. The patient showed a satisfactory response to oral Itraconazole over 12 weeks.

The co-culture strategy, which mimics natural ecology by constructing an artificial microbial community, is a useful tool to activate the biosynthetic gene clusters to generate new compounds. However, without optimization of fermentation conditions, the antagonism between the microbes often interferes with the production of secondary metabolites. In this study, the fermentation conditions of co-culture of Aspergillus sydowii and Bacillus subtilis were optimized by response surface methodology to increase the production of active metabolites against Staphylococcus aureus. After optimization, the inhibitory rate of the co-culture extract was 74.62%, which was 29.20% higher than that of the initial conditions. Meanwhile, a total of 15 newly biosynthesized metabolites were detected only in optimized co-culture, occupying 13.2% of all detected metabolites. The structures of the 12 metabolites with high variable importance in projection score were elucidated by the established LC-MS/MS approach integrated with various metabonomic tools. Among them, 7 metabolites were newly induced and the content of other 5 metabolites increased by 1.1-2.4 folds in optimized co-culture. The bioassay of metabolites in co-culture against S. aureus indicated that compounds (-)- (7S)- 10-hydroxysydonic acid, serine sydonate and macrolactin U' contributed much to the increment of antibacterial activity. This study demonstrated that optimizing the fermentation conditions of co-culture was beneficial to changing the metabolite profile and effective to induce the biosynthesis of active metabolites.

The cryptic Aspegillus species are rare, these microorganisms are usually more resistant to common antifungal therapies. Therefore, a correct identification is important when evaluating the impact of such species in aspergillosis.We aimed to describe the frequency, clinical and microbiological characteristics, and the outcomes of those cases of aspergillosis caused by cryptic species in a tertiary hospital.We retrospectively identified all microbiologically documented cases of aspergillosis between January 2013 and December 2018. Definitive species identification of clinically significant isolates was achieved via sequencing methods. The polymerase chain reaction (PCR) products were sequenced, and the results obtained were compared to sequences deposited in GenBank. Antifungal susceptibility testing was performed using the Sensititre® YeastOne® panel.A total of 679 Aspergillus isolates were recovered from 489 patients, of which 109 were clinically relevant. Ten (9.2%) isolates were identified as cryptic species: Aspergillus arcoverdensis (2), Aspergillus lentulus (2), Aspergillus ellipticus (2), Aspergillus alliaceus (1), Aspergillus nomius (1), Aspergillus tubingensis (1) and Aspergillus montevidensis (1). Most patients already suffered some type of immunosuppression. Half of these patients had required intensive care before the infection showed up, and most of them had a pulmonary infection. Mortality at the 100-day follow-up was 40%. Antifungal susceptibility testing was performed on three of the isolates (A. arcoverdensis, A. tubingensis and A. nomius), which showed high minimum inhibitory concentrations (MIC) for azoles and amphotericin B.The frequency of cryptic species in our centre was 9.2%. Most patients had some degree of immunosuppression, and the mortality rate was 40%.

Chronic pulmonary aspergillosis (CPA) is an infection of the lung usually caused by Aspergillus fumigatus in patients with pre-existing pulmonary diseases. Its diagnosis hinges on demonstrating IgG antibodies against A. fumigatus. Herein, we evaluated the performance of a newly introduced point of care test (POCT) kit, the LDBio Aspergillus IgG/IgM lateral flow assay (LFA) in India with the standard ImmunoCAP kit for diagnosing CPA.A total of 60 serum samples (30 CPA cases and 30 controls) were evaluated by the Aspergillus immunochromatographic test (ICT) IgG/IgM LFA. Fluorescent-enzyme immunoassay was used to determine specific A. fumigatus-IgG concentrations (positive27 mgA/L). Further, a systematic review and meta-analysis of studies (up to August 26, 2021) reporting the performance of LDBio ICT for the diagnosis of CPA was performed.A sensitivity of 86.7%, specificity of 90%, negative predictive value of 87.1%, positive predictive value of 89.7%, negative likelihood ratio of 0.15, positive likelihood ratio of 8.67, and was observed for the LDBio IC. There was good agreement between LDBio ICT and ImmunoCAP (88.3%) with a Cohen's Kappa score of 0.77. Our systematic review identified four studies and the pooled sensitivity of 90%, specificity of 91%, area under the curve of 0.94 and diagnostic odds ratio of 57.2, for CPA diagnosis by LDBio ICT.Aspergillus LDBio ICT assay exhibits good sensitivity and can be used to screen CPA cases.

Nuclear factor-kappa B (NF-κB) signaling is an intracellular signaling pathway involved in inflammatory responses and the pathogenesis of various cancers, including ependymoma, which is a rare and chemotherapy-resistant glioma. Several isoforms of fusion proteins that consist of a nuclear protein, zinc finger translocation associated (ZFTA), and RELA (ZFTA-RELA), an NF-κB-signaling effector transcription factor, cause excessive activation of the NF-κB signaling pathway and result in supratentorial ependymomas (ST-EPN-RELA). As inhibitors of NF-κB activity induced by ZFTA-RELA are expected to be therapeutic agents for ST-EPN-RELA, we established an NF-κB responsive luciferase reporter cell line that expresses the most common isoform of ZFTA-RELA in a doxycycline-dependent manner. Using this reporter cell line, we screened fungus extracts for compounds that inhibit the NF-κB activity induced by ZFTA-RELA expression and identified aszonalenin, an alkaloid from Aspergillus novofumigatus. We also purified analogs of aszonalenin, namely acetylaszonalenin and epi-aszonalenin B and C. In a luciferase assay using cells constitutively expressing luciferase (counter assay), acetylaszonalenin and epi-aszonalenin C showed non-specific inhibition of the luciferase activity. Aszonalenin and epi-aszonalenin B inhibited the NF-κB responsive luciferase activity by expressing ZFTA-RELA more strongly than the luciferase activity in the counter assay. The upregulation of endogenous NF-κB responsive genes, such as CCND1, ICAM1, and L1CAM, by ZFTA-RELA expression was inhibited by epi-aszonalenin B, but not by aszonalenin. This study suggests that epi-aszonalenin B may be a lead compound for the therapeutic development of ST-EPN-RELA.

Blue mold caused by Penicillium italicum is a severe postharvest disease in citrus fruits. In this study, the fermentation product (FP-E) of Aspergillus aculeatus GC-09, an endophytic fungus isolated from a citrus plant, was found to exhibit antifungal activity against P. italicum with a MIC of 0.3125 mg/mL. The fungus A. aculeatus GC-09 was identified based on the studies of morphology and ITS nucleotide sequence. FP-E significantly inhibited the spore germination and mycelial growth of P. italicum. Scanning electron microscopy (SEM) results of P. italicum treated with FP-E showed shrunken, distorted and collapsed hyphae and conidiospores, indicative of the cell membrane damage, which was further confirmed by the propidium iodide (PI) fluorescent staining analysis. Consistent with the microscopy observation, FP-E led to the leakage of cellular constituents from P. italicum, which is evident from the increase in electrical conductivity and nucleic acid contents in the mycelial solution incubated with FP-E. In addition, FP-E treatment considerably increased the intracellular reactive oxygen species (ROS) content, and reduced the enzyme activities of both catalase (CAT) and peroxidase (POD) in P. italicum cells. Furthermore, orange fruits treated with FP-E showed fewer disease symptoms compared to the untreated fruits. These results suggested that the antifungal activity of FP-E might be associated with the disruption of cell membrane integrity, the accumulation of ROS level, and the reduction of the antioxidant enzymes activity of P. italicum. Therefore, A. aculeatus GC-09 might be a potential microbial resource for the biocontrol of citrus postharvest blue mold.

Pulmonary oxalosis can be fatal, and Aspergillus tubingensis is commonly resistant to azoles in Japan. We report a case of bronchopulmonary oxalosis caused by A. tubingensis in a non-neutropenic patient who was successfully treated with voriconazole monotherapy. The susceptibility of the isolates to voriconazole and the effective elimination of contagious necrotic tissue by expectoration seemed to be two major factors contributing to the patient's survival. According to the literature review, pulmonary oxalosis is associated with a high mortality rate over a short term. An exploration of detailed information about the genomic characteristics and drug susceptibility of Aspergillus isolates is important for the development of treatment strategies for this life-threatening disease.

Due to the dissemination of airborne conidia and spores, molds can contaminate various surfaces. In the food industry sector, their presence and development can have health and economic implications. In order to control these undesirable microorganisms, various approaches can be used but the main one relies on the use of disinfectants. The objective of this review is to report the existing studies on the effect of various disinfectant molecules (i.e., sodium hypochlorite, chlorine dioxide, ethanol and other alcohols, hydrogen peroxide, peracetic acid, and quaternary ammonium compounds) on the inactivation of fungal spores. These studies were sorted depending on the targeted fungal species. Noteworthy, in the food industry, four log and three log reductions are required to claim a fungicidal activity for suspension (European Standard 1650, 2019) and surface (European Standard 13697/IN1, 2019) treatments, respectively. Most of the presented studies concerned Penicillium and Aspergillus species (44 and 31% of the literature, respectively). In general, for a given disinfection procedure, ascospores were more resistant than conidia, and Aspergillus conidia were more resistant than Penicillium ones. However, the variability of encountered molds (e.g. species, strains, physiological state) and disinfection procedures (e.g. molecules, concentrations, contact time) affected the efficacy of disinfectants.

We present the case of a previously healthy 54-year-old man who was hospitalized for an Aspergillus fumigatus infection of an open window thoracotomy. Patient was successfully treated for 8 consecutives weeks with daily topical pleural liposomal amphotericine B administered by soaked gauzes combined with systemic therapy.

Microbial food alterations lead to unfit products for consumption, and their discarding, to significant economic losses for the food industry. During storage, fresh foods offer available niches for the survival and growth of undesirable microorganisms. In dairy products, data regarding spoilage and/or pathogenic bacteria is better documented than those for molds and yeasts. Dairy products are less susceptible to mold's contamination than products such as fruits and vegetables, due to their refrigerated storage; their elaboration from heat-treated milk and, for fermented ones, the dominant microbiota that acidifies the medium. However, even cheeses and yogurts may be susceptible to mold contamination. Atypical cases of yogurt samples containing spoilage microorganisms not previously reported (molds producing gas and bacteria of the genus Gluconobacter) in Argentinean fermented milks are presented here. For yogurt, in particular, the “classic” altering organisms were always being yeasts, and in other countries, molds belonging to the genus Aspergillus. Fil: Capra, María Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina Fil: Frisón, Laura Noemí. Universidad Nacional del Litoral. Facultad de Ingeniería Química; Argentina Fil: Chiericatti, Carolina Andrea Antonia. Universidad Nacional del Litoral. Facultad de Ingeniería Química; Argentina Fil: Binetti, Ana Griselda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina Fil: Reinheimer, Jorge Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina

As the dominant fungus during the fermentation of Fuzhuan brick tea, Aspergillus cristatus is easily induced to undergo a sexual cycle under low-salt stress. However, the underlying regulatory mechanism of sexual reproduction is unclear. Here, we report a P53-like transcription factor AcndtA, which encodes an NDT80 DNA binding protein and regulates fungal reproduction, pigmentation and the stress response. Both insertion and deletion mutants of AcndtA exhibited a complete blockade of cleistothecium formation, and overexpressing AcndtA strains (OE: AcndtA) exhibited significantly reduced cleistothecium production, indicating that AcndtA plays a vital role in sexual development. Osmotic stress tests showed that overexpression of AcndtA had a negative impact on growth and conidia production. Additionally, AcndtA insertion, deletion and overexpression mutants exhibited reduced pigment formation. All the above developmental defects were reversed by the re-introduction of the AcndtA gene in ΔAcndtA. Moreover, the growth of AcndtA mutants in carbon-limited medium was better than that of the WT and OE: AcndtA strains, indicating that AcndtA is involved in carbon metabolism. Transcriptional profiling data showed that AcndtA regulated the expression of several genes related to development, osmotic stress and carbon metabolism.

Filamentous fungi Aspergillus and Fusarium species are major causes of visual impairment and blindness in immune competent individuals. Once conidia penetrate the corneal epithelium and enter the stroma, they undergo germination, and exposure of cell wall components induces a pronounced neutrophil-rich cellular infiltrate. In this review, we discuss Aspergillus and novel Fusarium virulence factors that are required for corneal infection, and describe the multiple functions of neutrophils in limiting hyphal growth in the cornea. This review will also discuss the role of neutrophils as an important source of cytokines in fungal keratitis, and highlight recent studies identifying unique characteristics of neutrophil secretion of IL-1α and IL-1β.

Biodeterioration is related to several biological systems and metabolic activities of fungi and bacteria. Fungal contamination is determined by the availability of water and temperature. Aspergillus is a fungus with great enzymatic capacity and with a high biodeterioration power. For this reason, it represents a threat to the conservation of cultural heritage objects. In this work, an exhaustive search of all the species involved was carried out, describing in detail the deterioration produced in different objects (wooden objects, glasses, paper, paintings, textiles, audiovisuals, glasses, human remains) as well preventive treatments. Like an opportunistic fungus it might cause some illness related to workers, a situation that is also addressed. Since the number of species increased from 180 to over 400 in the last 20 years, a taxonomy update has been done.

The CRISPRoff system was recently introduced as a programmable epigenetic memory writer that can be used to silence genes in human cells. The system makes use of a dead Cas9 protein (dCas9) that is fused with the ZNF10 KRAB, Dnmt3A, and Dnmt3L protein domains. The DNA methylation resulting from the CRISPRoff system can be removed by the CRISPRon system that consists of dCas9 fused to the catalytic domain of Tet1. Here, the CRISPRoff and CRISPRon systems were applied for the first time in a fungus. The CRISPRoff system resulted in an inactivation up to 100 % of the target genes flbA and GFP in Aspergillus niger. Phenotypes correlated with the degree of gene silencing in the transformants and were stable when going through a conidiation cycle, even when the CRISPRoff plasmid was removed from the flbA silenced strain. Introducing the CRISPRon system in a strain in which the CRISPRoff plasmid was removed fully reactivated flbA showing a phenotype similar to that of the wildtype. Together, the CRISPRoff and CRISPRon systems can be used to study gene function in A. niger.

Aspergillus neoniger NCIM 1400 whose cell-free fraction was earlier established for transglycosylation activity conferred by α-glucosidase gene (agdA), was subjected to sequence analysis. Preliminary results revealed certain dynamics in the intron splicing mechanism, and to ascertain these molecular events, a detailed study was carried. The electrophoresis results from the cDNA portion (B-fragment) of agdA showed multiple bands, indicating the amplification of one or more fragments. The sequence results of cDNA cloned vector revealed the retention type of alternative splicing in the agdA. The splicing mechanism of agdA in NCIM 1400 was compared to different A. niger strains, which harbours agdA orthologues, using PCR. It was observed that effective intron splicing leads to higher α-glucosidase activity from these selected Aspergillus spp. To explore the dynamics of intron retention in A. neoniger NCIM 1400, time-course analysis of intron retention, enzyme activity, and sugar consumption were carried over a period of 168 h of fungal growth. RT-qPCR results revealed that introns retention was not detected during the initial growth phase when the maltose and its hydrolysed product, glucose were consumed. Here we demonstrate that exhaustion of maltose causes increase in retention of introns in the mRNA transcripts of agdA gene, and this could be the possible mode of regulating this gene.

Summary The objective of this study was to review the antifungal susceptibility of clinical mould isolates performed by the New Zealand Mycology Reference Laboratory. Isolates were either local or referred for testing from other New Zealand laboratories. All isolates were tested by the broth colorimetric microdilution method, Sensititre YeastOne (SYO). Epidemiological cut-off values (ECVs) derived from either the Clinical and Laboratory Standards Institute (CLSI) method or SYO were used to determine the proportion of non-wild type (non-WT) isolates, i.e., those with an increased likelihood to harbour acquired mechanisms of resistance. A total of 614 isolates were tested. Most isolates (55%) were from the respiratory tract followed by musculoskeletal tissue (17%), eye (10%) and abdomen (5%). The azoles had similar activity except for voriconazole which was less active against the Mucorales. The echinocandins had good activity against Aspergillus spp., other hyaline moulds and dematiaceous isolates but were inactive against Fusarium spp., Lomentospora prolificans and the Mucorales. Amphotericin B had best activity against the Mucorales. The two least susceptible groups were Fusarium spp. and L. prolificans isolates. Three Aspergillus isolates were non-WT for amphotericin B, and four non-WT for azoles. Non-WT were not encountered for caspofungin. Non-Aspergillus isolates in New Zealand have susceptibility patterns similar to those reported elsewhere. In contrast to a growing number of other countries, azole resistance was rare in A. fumigatus sensu stricto. Non-WT isolates were uncommon. The results provide a baseline for monitoring emerging antifungal resistance in New Zealand and support current Australasian treatment guidelines for invasive fungal infections.

Many genes responsible for melanin biosynthesis in fungi were physically linked together. The PKS gene clusters in most of the melanin-producing fungi were regulated by the Cmr1. It was found that a close rearrangement of the PKS gene clusters had evolved in most of the melanin-producing fungi and various functions of melanin in them were beneficial to their adaptation to the changing environments. The melanin-producing fungi had undergone at least five large-scale differentiations, making their PKS gene clusters be quickly evolved and the fungi be adapted to different harsh environments. The recent gene losses and expansion were remarkably frequent in the PKS gene clusters, leading to their rapid evolution and adaptation of their hosts to different environments. The PKS gene and the CMR1 gene in them were subject to a strong co-evolution, but the horizontal gene transfer events might have occurred in the genome-duplicated species, Aspergillus and Penicillium.

A study was carried out to investigate the characterization of a novel Aspergillus sulphureus JCM01963 xylanase (AS-xyn10A) with a carbohydrate binding module (CBM) and its application in degrading alkali pretreated corncob, rapeseed meal and corn stover alone and in combination with a commercial cellulase. In this study, the 3D structure of AS-xyn10A, which contained a CBM at C-terminal. AS-xyn10A and its CBM-truncated variant (AS-xyn10A-dC) was codon-optimized and over-expressed in Komagaella phaffii X-33 (syn. Pichia pastoris) and characterized with optimal condition at 70 °C and pH 5.0, respectively. AS-xyn10A displayed high activity to xylan extracted from corn stover, corncob, and rapeseed meal. The concentration of hydrolyzed xylo-oligosaccharides (XOSs) reached 1592.26 μg/mL, 1149.92 μg/mL, and 621.86 μg/mL, respectively. Xylobiose was the main product (~70%) in the hydrolysis mixture. AS-xyn10A significantly synergized with cellulase to improve the hydrolysis efficiency of corn stover, corncob, and rapeseed meal to glucose. The degree of synergy (DS) was 1.32, 1.31, and 1.30, respectively. Simultaneously, XOSs hydrolyzed with AS-xyn10A and cellulase was improved by 46.48%, 66.13% and 141.45%, respectively. In addition, CBM variant decreased the yields of xylo-oligosaccharide and glucose in rapeseed meal degradation. This study provided a novel GH10 endo-xylanase, which has potential applications in hydrolysis of biomass.

The mycotoxin patulin is primarily produced as a secondary metabolite by numerous fungal species and predominantly by Aspergillus, Byssochlamys, and Penicillium species. It is generally associated with fungal infected food materials. Penicillium expansum is considered the only fungal species liable for patulin contamination in pome fruits, especially in apples and apple-based products. This toxin in food poses serious health concerns and economic threat, which has aroused the need to adopt effective detection and mitigation strategies. Understanding its origin sources and biosynthetic mechanism stands essential for efficiently designing a management strategy against this fungal contamination. This review aims to present an updated outline of the sources of patulin occurrence in different foods and their biosynthetic mechanisms. It further provides information regarding the detrimental effects of patulin on human and agriculture as well as its effective detection, management, and control strategies.

The increased availability and production of lignocellulosic agroindustrial wastes has originated proposals for their use as raw material to obtain biofuels (ethanol and biodiesel) or derived products. However, for biomass generated from lignocellulosic residues to be successfully degraded, in most cases it requires a physical (thermal), chemical, or enzymatic pretreatment before the application of microbial or enzymatic fermentation technologies (biocatalysis). In the context of enzymatic technologies, fungi have demonstrated to produce enzymes capable of degrading polysaccharides like cellulose, hemicelluloses and pectin. Because of this ability for degrading lignocellulosic material, researchers are making efforts to isolate and identify fungal enzymes that could have a better activity for the degradation of plant cell walls and agroindustrial biomass. We performed an in silico analysis of alpha-glucoronidase in 82 accessions of the genus Aspergillus. The constructed dendrograms of amino acid sequences defined the formation of 6 groups (I, II, III, IV, V, and VI), which demonstrates the high diversity of the enzyme. Despite this ample divergence between enzyme groups, our 3D structure modeling showed both conservation and differences in amino acid residues participating in enzyme-substrate binding, which indicates the possibility that some enzymes are functionally specialized for the specific degradation of a substrate depending on the genetics of each species in the genus and the condition of the habitat where they evolved. The identification of alpha-glucuronidase isoenzymes would allow future use of genetic engineering and biocatalysis technologies aimed at specific production of the enzyme for its use in biotransformation.

Lung granulomas are uncommon in Thailand. The disease typically develops from an occupational environment and is mostly caused by infection. Herein is a case report of a female patient, aged 48, working as a nurse in an Accident and Emergency Department at a hospital. Eighteen years prior to admission the patient was diagnosed with myasthenia gravis and pulmonary tuberculosis. The chest X-ray and CT scans showed a solitary pulmonary nodule in the lower left lung. The patient received an open thoracotomy with a left lobectomy. Granulomatous and nonseptate hyphae were found in the pathology diagnosis. The patient was thus diagnosed as having a lung granuloma. The galactomannan antigen test was positive. The solitary pulmonary nodule—found from the use of a Polymerase Chain Reaction (PCR) test—was an Aspergillus spp. The fungus culture was collected from air samples. The air samples were collected by the impaction technique using a microbial air sampler. Three types of Aspergillus spp. were found as well as Penicillium spp. and Monilia sitophila. The Aspergillus spp. was a match for the patient's disease. The patient was diagnosed as having a lung granuloma possibly Aspergillus nodule which was caused by airborne Aspergillus spp. from the occupational environment.

Keratomycosis or mycotic keratitis is recognized as one of the major causes of ophthalmic morbidity worldwide. The most common organisms linked to keratomycosis include Candida spp., Fusarium spp., and Aspergillus spp. However, varieties of saprobic fungi have been reported as causative agents of keratomycosis. Amongst these are members of the genus Colletotrichum. Herein we present the first reported case of C. chlorophyti infection in a post-corneal transplant patient, suggesting an increasing role for Colletotrichum species as emerging human pathogens, particularly in the transplant population.

A new azaphthalide derivative, (S)-3-hydroxy-2,7-dimethylfuro [3,4-b]pyridin-5(7H)-one (1), and a new phthalide derivative, (S)-7-hydroxy-3-((S)-1-hydroxyethyl)isobenzofuran-1(3H)-one (2), were isolated from the coral-derived fungus Aspergillus sp. SCSIO41405, together with eight known compounds (3–10). Their structures, including absolute configurations, were determined by NMR and MS spectroscopic analyses, optical rotation, calculated electronic circular dichroism (ECD) and X-ray crystal data. The isolated compounds were examined for antibacterial and enzyme inhibition activities. Compounds 7 and 8 exhibited weak antibacterial activities, and 10 displayed moderate inhibition on pancreatic lipase with an IC50 value of 15.6 μg/mL.

Aspergillus species are ubiquitous in the environment. Aspergillosis is acquired by inhalation of Aspergillus spores. In normal hosts, spore inhalation rarely causes lung disease. Pulmonary Aspergillosis covers a wide spectrum of clinical syndromes depending on the interaction between Aspergillus and the host (immune-status, prior bronchopulmonary disease). It runs the gamut from invasive Aspergillosis to Aspergillus bronchitis. Invasive Aspergillosis usually occurs in severely immunocompromised patients, typically in neutropenic but also in non-neutropenic patients. Chronic pulmonary Aspergillosis affects patients with chronic structural lung disease such as COPD or previous mycobacterial lung disease, but without other significant immunocompromise. Aspergillus bronchitis affects patients with bronchial disease such as bronchiectasis. Allergic bronchopulmonary Aspergillosis affects patients with bronchial asthma or cystic fibrosis, and is due to an allergic response to Aspergillus.

To improve utilization of American-ginseng-leaf ginsenosides, the PPD-ginsenosides from American-ginseng-leaves was reacted by a ginsenosidase type-I from Aspergillus sp.agl-84 to prepare minor ginsenosides C-K and C-Mx. PPD-ginsenosides, containing over 79 % of Rb3 and Rd, were separated from American-ginseng-leaves; the yield was 10.5 %. The molecular weight of ginsenosidase type-I from Aspergillus sp.agl-84 was approximately 75.2 kDa; Michaelis constants Km and Vmax were 14.8 mM and 58.6 mM/h for Rb3, 15.4 mM and 56.8 mM/h for Rd. The enzyme reaction velocities on 3-O-glucosides of Rb3 and Rd were higher than that of other Aspergillus strain ginsenosidase type-I to prove that the enzyme was more suitable for conversion of PPD-ginsenosides from American-ginseng-leaves into C-K and C-Mx. According to enzyme reaction dynamic, 2.5 % PPD-ginsenosides were reacted at 45 °C and pH 5.0 for 24 h by a crude ginsenosidase type-I from Aspergillus sp.agl-84 strain. Here, 4.1 g C-K and 4. 2 g C-Mx were obtained from 20 g PPD-ginsenosides separating from 190 g American-ginseng-leaves. C-K molar yield was 34.3 %, C-Mx molar yield was 29.0 %, the total molar yield was 63.3 %. Therefore, minor ginsenosides C-K and C-Mx were successfully prepared from PPD-ginsenosides of American-ginseng-leaves using crude enzyme from Aspergillus sp.agl-84 at low cost.

Clinical mycology is in continuous development. The appearance of new clinical guidelines has made it possible to improve the approach to opportunistic fungal infections, especially in immunosuppressed patients (oncohematological and/or transplant recipients). At the same time, the development of new diagnostic tools and new antifungals with a greater spectrum of action and fewer side effects have led to faster diagnoses and treatments that are more effective. Along with these advances, there has been a change in the epidemiology of invasive fungal infection (IFI), with the appearance of new patients (e.g., COPD, liver cirrhosis, post-influenza) and new microorganisms (Candida auris, Lomentospora prolificans, mucorales), and resistant fungi (isolates of Aspergillus resistant to azoles) which the clinician must take into account when choosing the treatment of a patient with an IFI. In this paper we will briefly review the advances in recent decades and the emerging problems.

Los herbicidas a base de glifosato (HBG) son los más aplicados a nivel mundial en la producción de maíz. La aparición de malezas resistentes a glifosato condujo a la aplicación repetida de altas dosis. Además de las condiciones ambientales, la presencia de HBG afectan el desarrollo de especies de Aspergillus y la producción de aflatoxina B1(AFB1). El objetivo de este trabajo fue evaluar la influencia de un HBG comercial en el crecimiento de cepas de Aspergillus flavus y Aspergillus parasiticus y su producción de AFB1 en granos de maíz, bajo diferentes condiciones de actividad de agua (aW). Se evaluaron las siguientes concentraciones de ingrediente activo glifosato: 20, 50, 200 y 500 mM. La fase de latencia y la velocidad decrecimiento no se modificaron con 20 y 50 mM (dosis recomendadas a campo) a 0,98 y 0,95 de aW; mientras que el crecimiento disminuyó en todas las condiciones de aW cuando las concentraciones de HBG fueron de 200 y 500 mM. En general, cuando aumentó la concentración de HBG, disminuyó la producción de AFB1. Hubo un aumento significativo en la acumulación de toxina solo en una condición de aW (0,95) a los 21 días con 50 mM de HBG en A. flavus, y con 20 y 50 mM en A. parasiticus. Estos resultados muestran que concentraciones de HBG similares a las dosis recomendadas para uso a campo no incrementan el crecimiento de Aspergillus sección Flavi en granos de maíz, pero sí la producción de AFB1, bajo condiciones favorables de disponibilidad de agua y temperatura. Glyphosate-based herbicides (GBH) are the main pesticides applied worldwide on maize production. Glyphosate-resistant weeds led to the repeated application of high doses of the pesticide. In addition to environmental conditions, the presence of GBH affects the development of Aspergillus species and aflatoxin B1(AFB1) production under in vitro conditions. The aim of this work was to evaluate the influence of a commercial GBH on growth and AFB1 production by Aspergillus flavus and Aspergillus parasiticus strains under different water activity (aW) conditions. The following concentrations of active ingredient glyphosate were evaluated:20, 50, 200 and 500 mM. The lag phase prior to growth and growth rate did not change at 20 and 50 mM (that is, at field recommended doses) at 0.98 and 0.95 aW; however, at increasing GBH concentrations, between 200 and 500 mM, the growth rate decreased at all aW conditions. In general, as the GBH concentration increased, AFB1production decreased. However, a significant increase in toxin accumulation was found only at one of the aW conditions (0.95) at 21days with 50 mM of GBH in A. flavus and 20 and 50 mM of GBH in A. parasiticus. These results show that, even though Aspergillus section Flavi growth did not increase, AFB1 production increased on maize grains at GBH concentrations similar to those of field recommended doses under favorable water availability and temperature conditions. Fil: Benito, Nicolas. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Magnoli, Karen. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Carranza, Cecilia Soledad. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Aluffi, Melisa Egle. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Magnoli, Carina Elizabeth. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Barberis, Carla Lorena. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina

The essential oil Pickering emulsions stabilized by zein and chitosan nanoparticles (ZCPs) were investigated. ZCP stabilized Pickering emulsions encapsulating citral and/or cinnamaldehyde (ZCP-CT, ZCP-CN, ZCP-CTCN) were constructed as an effective antimicrobial system, and the stability and antifungal properties were evaluated. When the ratio of zein to chitosan was 20:1, the emulsion showed good stability within 15 days and good sustained release ability during 9-day storage experiment. All Pickering emulsions could reduce fungal growth and ochratoxin A production. The ZCP-CN inhibited 62.3 ± 0.5 % Aspergillus westerdijkiae and reduced mycotoxin production by 83.8 ± 1.0 % in corn within 15 days. Meanwhile, the ZCP-CTCN inhibited 58.6 ± 1.0 % Aspergillus carbonarius within 8 days, and the inhibition rate of mycotoxin was up to 99.8 ± 0.3 % in grape. The Pickering emulsions could extend the release time of essential oil and improve the anti-fungal efficiency, which provide new ideas for the development of new food mildew inhibitor.

Invasive aspergillosis (IA) affects more than 300,000 people annually worldwide with a case fatality rate reaching 80%. However, in Africa despite the presence of risk factors for the development of IA, the burden of these fungal infections remained unknown. This systematic review aimed to update the available information on the epidemiology and the therapeutic management of IA in Africa. The published papers were systematically searched on major medical databases from September 20 to October 10, 2021. The list of references of eligible articles and the Google scholar database were also checked in order to search for possible eligible articles. Results were reported following the Preferred Reporting Items for Systematic and Meta-analyses (PRISMA) guidelines. The search yielded 1864 articles of which 29 met the inclusion criteria. This systematic review showed the existence of IA in Africa. The prevalence of IA can reach 27% with a fatality rate of more than 60%. The most common clinical form of IA found was invasive pulmonary aspergillosis. The main predisposing conditions identified were neutropenia, HIV/AIDS, renal transplant recipients, and renal failure. Aspergillus section Flavi and Nigri were the main Aspergillus species identified and Aspergillus section Fumigati was uncommon. The main management strategy for IA cases was to start antifungal therapy only after a failure of broad-spectrum antibiotic therapy. This review provided evidence of the existence of invasive aspergillosis in Africa and especially a high rate of undiagnosed invasive aspergillosis cases.

Bacterial or virus co-infections with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been reported in many studies, however, the knowledge on Aspergillus co-infection among patients with coronavirus disease 2019 (COVID-19) was limited. This literature review aims to explore and describe the updated information about COVID-19 associated with pulmonary aspergillosis. We found that Aspergillus spp. can cause co-infections in patients with COVID-19, especially in severe/critical illness. The incidence of IPA in COVID-19 ranged from 19.6% to 33.3%. Acute respiratory distress syndrome requiring mechanical ventilation was the common complications, and the overall mortality was high, which could be up to 64.7% (n = 22) in the pooled analysis of 34 reported cases. The conventional risk factors of invasive aspergillosis were not common among these specific populations. Fungus culture and galactomannan test, especially from respiratory specimens could help early diagnosis. Aspergillus fumigatus was the most common species causing co-infection in COVID-19 patients, followed by Aspergillus flavus. Although voriconazole is the recommended anti-Aspergillus agent and also the most commonly used antifungal agent, aspergillosis caused by azole-resistant Aspergillus is also possible. Additionally, voriconazole should be used carefully in the concern of complicated drug-drug interaction and enhancing cardiovascular toxicity on anti-SARS-CoV-2 agents. Finally, this review suggests that clinicians should keep alerting the possible occurrence of pulmonary aspergillosis in severe/critical COVID-19 patients, and aggressively microbiologic study in addition to SARS-CoV-2 via respiratory specimens should be indicated.

Only a few known epoxide hydrolases (EHs) displayed activity towards o-nitrostyrene oxide (4a), presumably owing to the large steric hindrance caused by o-nitro substituent. Therefore, excavating EHs with high activity and enantio- and/or regio-selectivity towards racemic (rac-) 4a is essential but challenging. Here, AuEH2 from Aspergillus usamii was expressed in E. coli BL21(DE3). E. coli/Aueh2, an E. coli transformant expressing AuEH2, possessed EH activities of 16.2–184 U/g wet cell towards rac-styrene oxide (1a) and its derivatives (2a–13a), and the largest enantiomeric ratio of 96 towards rac-4a. The regioselectivity coefficients, βR and βS, of AuEH2 were determined to be 99.2% and 98.9%, suggesting that it regiopreferentially attacks the Cβ in the oxirane rings of (R)- and (S)-4a. Then, the nearly perfect kinetic resolution of 20 mM rac-4a in pure water was carried out using 20 mg/mL wet cells of E. coli/Aueh2 at 25 °C for 50 min, retaining (S)-4a with over 99% ees and 48.9% yields, while producing (R)-o-nitrophenyl-1,2-ethanediol (4b) with 95.3% eep and 49.8% yieldp. To elucidate the molecular mechanism of AuEH2 with high enantiopreference for (R)-4a, its crystal structure was solved by X-ray diffraction and the molecular docking of AuEH2 with (R)- or (S)-4a was simulated.

Crude oil biodegrading microorganism considers the key role for environmental preserving. In this investigation, crude oil biodegrading fungal strains have been isolated in polluted soil of crude-oil at khurais oil ground in Kingdom of Saudi Arabia. Among of 22 fungal isolates, only three isolates reflected potential capability for oil degradation. These isolates were identified and submitted to GenBank as (A1) Aspergillus polyporicola (MT448790), (A2) Aspergillus spelaeus (MT448791) and (A3) Aspergillus niger (MT459302) through internal-transcribed spacer-regions (ITS1&ITS2) for sequencing in molecular marker. Comparing with controls, strain (A1) Aspergillus niger was superior for biodegradation ability (58%) comparing with Aspergillus polyporicola and Aspergillus spelaeus degrading were showed 47 and 51 % respectively. Employed CO2 evolution as indicator for petroleum oil biodegradation by the fungal isolates reflected that, Aspergillus niger emission highest CO2 (28.6%) comparing with Aspergillus spelaeus and Aspergillus polyporicola which showed 13 % and 12.4 % respectively. capability of Aspergillus sp. to tolerate and adapted oil pollutants with successful growth rate on them, indicated that it can be employed as mycoremediation agent for recovering restoring ecosystem when contaminated by crude oil.

The stability of asbestos in natural environment with its biological aggressiveness and toxicity leading to carcinogenesis is related to its fibrous structure, dimensions and presence of iron in its chemical composition. In an effort to reduce its toxicity by limiting its biopersistence, physico-chemical methods have proved futile. A probable solution highlighted in the present investigation is bioremediation. Capitalizing the iron chelating ability of siderophores, as a tool for reducing asbestos toxicity is the main feature under study. A comparative assessment of the siderophore synthesizing ability and their iron chelation potential showed that the identified bacterial species namely Bacillus subtilis and Bacillus atrophaeus as well as the fungal species Aspergillus tubingenesis and Coemansia reversa harboured a similar effectivity towards reducing asbestos toxicity. A microcosm approach was thus envisaged as a probable tool for the same.

Rice is a staple food consumed across the world and its purity is essential to manage human health. However, fungal mycotoxins are one of the real challenges for maintaining the purity of rice intended for human consumption. The present study was designed to detect the fungal isolates present in the imported rice cultivars obtained from the local markets of the Kingdom of Saudi Arabia and the mycotoxins they produce using molecular approaches. The morphological analysis and PCR-based sequencing of ITS-rDNA specify that the isolates in the contaminated rice belonged to Aspergillus and Penicillium genera. The phylogenetic analysis based on sequences of ITS-rDNA regions of the isolates confirmed that the Aspergillus isolates were homological to A. flavus and A. nidulans. Whereas, the Penicillium isolates were homological to P. commune and P. chrysogenum. The identification of mycotoxins using LC-MS revealed that the isolates of the genus Aspergillus produced aflatoxin and ochratoxin. The isolates of Penicillium genus produced citreoviridin and citrinin. The levels of mycotoxins varied among each variety of the rice samples tested in total. Ochratoxin was produced at the least concentration (184.7 µg/kg) in the contaminated samples. Citrinin (23170.5 µg/kg) was produced at higher volumes in comparison to other mycotoxins. These levels of mycotoxin contamination are alarming and raise significant queries regarding the purity of the imported rice grains throughout the period of harvest and storage. Therefore, the development of practical control is required for consumer safety.

Invasive aspergillosis (IA) is associated with a high mortality rate in kidney-transplant recipients. Azole-resistance is increasing in Aspergillus fumigatus. We report a clinical case of a kidney-transplant recipient with cerebellar and pulmonary aspergillosis caused by azole-resistant Aspergillus parafelis (molecular identification through β-tubulin sequence). The patient experienced an effective resolution after three surgical procedures and associated antifungal therapy. This case highlights that azole-resistant aspergillosis should be considered in every patient with IA as long as susceptibility testing results are not known. Therefore, in selected patients with IA and central nervous system involvement, empirical combination antifungal therapy could be considered. Sí

Background Voriconazole (VRC) is widely recommended as the first-line therapy for invasive aspergillosis. However, surveillance studies have demonstrated that there is an increase in the frequency of azole resistance among Aspergillus fumigates isolates. In recent years, more studies on effective synergisms between natural agents and antifungal drugs have been published. Aims To evaluate the synergistic antifungal effect of glabridin (Gla) and VRC against A. fumigatus isolates. Methods Potential interactions between Gla and VRC were studied by using a microdilution checkerboard method based on the CLSI reference technique. To assess the interaction of drugs the fractional inhibitory concentration index (FICI) was calculated based on the Loewe Additivity model. Results The minimum inhibitory concentrations (MIC) obtained with Gla alone were relatively high (MIC50 16 μg/ml). However, our results showed synergistic interaction between Gla and VRC against A. fumigatus strains, with FICI range values between 0.15 and 0.5. Conclusions Synergistic activity of Gla and VRC against both VRC-sensitive and -resistant A. fumigatus isolates may lead to design new antifungal agents, especially for inhibiting those azole-resistant strains.

Utilization of agricultural wastes as cheap natural resources for production of bioactive products is currently attracting global attention. For this purpose, this study focused on isolation of Aspergillus wewitschiae MN056175 as promising producer of inulinase, then investigating physiochemical, kinetics and thermodynamics of the obtained inulinase, and its ability to extract bioactive fructo-oligosaccharides (FOS) from Cynara scolymus leaves (artichoke leaves, AL). A. wewitschiae MN056175 inulinase gave the maximum activity at temperature 60 °C and inulin concentration 1%. The kinetics including Km and Vmax were determined to be 105.26 mg·ml−1 and 83.33 μmol·ml−1·min−1, respectively. The thermodynamics including, Ea (activation energy) and Ed (activation energy for denaturation) were determined to be 21.82 and 73.21 kJ·mol−1, Kd, T1/2, D-value, ΔH°, ΔG° and ΔS° at 40, 50 and 60 °C which indicated the stability of A. wewitschiae MN056175 inulinase. Moreover, this inulinase was capable of hydrolyzing Cynara scolymus leaves into reducing sugar and 15 FOS with different DP, total carbohydrate, and protein content under different conditions designed by central composite design (CCD). The 15 AL FOS showed different high antioxidant and prebiotic activities. Central FOS with probiotic bacteria exhibited significant antimicrobial activity against tested gram positive bacteria in a way higher than those recorded against gram negative bacteria.

BACKGROUND MicroRNAs (miRNAs) are small non-coding RNAs of ∼22 nucleotides that play a crucial role in post-transcriptional regulation of gene expression. Dysregulation of miRNA expression has been shown during microbial infections. We sought to identify miRNAs that distinguish invasive aspergillosis (IA) from non-IA in lung transplant recipients (LTRs). METHODS We used NanoString nCounter Human miRNA, version 3, panel to measure miRNAs in bronchoalveolar lavage (BAL) samples from LTRs with Aspergillus colonization (ASP group) (n = 10), those with Aspergillus colonization and chronic lung allograft dysfunction (CLAD) (ASPCLAD group) (n = 7), those with IA without CLAD (IA group) (n = 10), those who developed IA with CLAD (IACLAD group) (n = 9), and control patients (controls) (n = 9). The miRNA profile was compared using the permutation test of 100,000 trials for each of the comparisons. We used mirDIP to obtain their gene targets and pathDIP to determine the pathway enrichment. RESULTS We performed pairwise comparisons between patient groups to identify differentially expressed miRNAs. A total of 5 miRNAs were found to be specific to IA, including 4 (miR-145-5p, miR-424-5p, miR-99b-5p, and miR-4488) that were upregulated and the pair (miR-4454 + miR-7975) that was downregulated in IA group vs controls. The expression change for these miRNAs was specific to patients with IA; they were not significantly differentiated between IACLAD and IA groups. Signaling pathways associated with an immunologic response to IA were found to be significantly enriched. CONCLUSIONS We report a set of 5 differentially expressed miRNAs in the BAL of LTRs with IA that might help in the development of diagnostic and prognostic tools for IA in LTRs. However, further investigation is needed in a larger cohort to validate the findings.

The fungal α-glucan synthases (Agss) are multi-domain proteins catalyzing biosynthesis of cell wall α-1,3-glucan which determines cell wall integrity or fungal pathogenicity and pullulan which is a maltotriosyl polymer made of α-1,4 and α-1,6 bound glucose units. The Agss family can be divided into 11 groups, some of which lost the original functions due to accumulation of harmful mutations or gene loss. Schizosaccharomyces pombe kept five kinds of Agss in the genome while Aspergillus spp. and Penicillium spp. lost one or two or three kinds of Agss. All the human, animal and plant pathogens kept only one single kind of Ags or only one active Ags for synthesis of cell wall α-1,3-glucan, a virulence factor. While the genus Aureobasidium spp. contained three kinds of Agss, of which only some of the Ags2 was involved in pullulan biosynthesis. Although many Agss contained Big_5 domain, only the Big_5 domain with conserved amino acids LQS from some strains of A. melanogenum could catalyze pullulan biosynthesis. This whole amino acid sequence and phylogenetic differences may cause non-α-1,3-glucan synthesizing activity of some fungal Agss.

The growing global awareness for environmental protection has inspired the exploration on producing active packaging films from bio-based materials. In present work, three types of active agents were studied by incorporating thymol(T), kesum(K), and curry(C) (10% wt.) into polylactic acid (PLA) to produce PLA-10T, PLA-10K, and PLA10-C packaging films via solvent casting method. The morphology, functional chemistry, thermal stability, permeability, and antimicrobial properties were evaluated for PLA films. Functional chemical analysis confirmed the presence of interfacial bonding between aromatic groups of active agents and PLA carbonyl group. PLA-10K exhibited the highest thermal resistance comparing to PLA-10T and PLA-10C while water vapor barrier was enhanced after incorporation of active agents. Qualitative observation had indicated that chicken meat could be preserved in the active films until 15 days, while odourless and firm texture properties retained in food sample. For disc diffusion assay (in vitro), it showed positive results against Gram-positive bacteria (Staphylococcus aureus) whereas with negative results against Gram-negative bacteria (Escherichia coli) and Aspergillus Brasiliensis due to embedded active agents within PLA matrix. We concluded that produced active agents filled PLA films potential to use in food packaging application to enhance the shelf life of meats, fruits and vegetables product.