Biodegradation of high molecular weight polycyclic aromatic hydrocarbons by Sarocladium terricola strain PYR-233 isolated from petrochemical contaminated sediment.

Polycyclic aromatic hydrocarbons (PAHs) were frequently found in sediment and were primarily treated through microbial degradation. Thus, efficient management of PAH pollution requires exploring the molecular degradation mechanisms of PAHs and expanding the pool of available microbial resources. A fungus (identified as Sarocladium terricola strain RCEF778) with the remarkable ability to degrade pyrene was screened from sediment near a petrochemical plant, and its growth and pyrene degradation characteristics were comprehensively investigated. The results showed that the fungus exhibited great effectiveness in pyrene degradation, with a degradation ratio of 88.97% at 21 days at the conditions: 35 °C, pH 7, 10 mg L-1 initially pyrene concentration, 3% supplementary salt, and glucose supplementation. The generation and concentration variation of the intermediate products were identified, and the results revealed that the fungus degraded pyrene through two pathways: by salicylic acid and by phthalic acid. Three sediments (M1, M2, M3), each exhibiting different levels of PAH pollution, were employed to examine the effectiveness of fungal degradation of PAHs in practical sediment samples. These data showed that with the fungus, the degradation ratios ranged from 13.64% to 23.50% for 2-3 rings PAHs, 40.93%-49.41% for 4 rings PAHs, and 39.59%-48.07% for 5-6 rings PAHs, which were significantly higher than those for the sediment without the fungus and confirmed the excellent performance of the fungal. Moreover, the Gompertz model was employed to analyze the degradation kinetics of 4-rings and 5-6 rings PAHs in these sediments, and the results demonstrated that the addition of the fungus could significantly increase the maximum degradation ratio, degradation start-up rate and maximum degradation rate of 4-rings and 5-6 rings PAHs and shorten the time required to reach the maximum degradation rate. This study not only supplied fungal materials but also established crucial theoretical foundations for the development of bioremediation technologies aimed at high molecular weight PAH-contaminated sediments.

Untargeted metabolomics and molecular docking studies on green silver nanoparticles synthesized by Sarocladium subulatum: Exploring antibacterial and antioxidant properties.

The biological synthesis of silver nanoparticles (Ag-NPs) with fungi has shown promising results in antibacterial and antioxidant properties. Fungi generate metabolites (both primary and secondary) and proteins, which aid in the formation of metal nanoparticles as reducing or capping agents. While several studies have been conducted on the biological production of Ag-NPs, the exact mechanisms still need to be clarified. In this study, Ag-NPs are synthesized greenly using an unstudied fungal strain, Sarocladium subulatum AS4D. Three silver salts were used to synthesize the Ag-NPs for the first time, optimized using a cell-free extract (CFE) strategy. Additionally, these NPs were assessed for their antimicrobial and antioxidant properties. Various spectroscopic and microscopy techniques were utilized to confirm Ag-NP formation and analyze their morphology, crystalline properties, functional groups, size, stability, and concentrations. Untargeted metabolomics and proteome disruption were employed to explore the synthesis mechanism. Computational tools were applied to predict metabolite toxicity and antibacterial activity. The study identified 40 fungal metabolites capable of reducing silver ions, with COOH and OH functional groups playing a pivotal role. The silver salt type impacted the NPs' size and stability, with sizes ranging from 40 to 52 nm and zeta potentials from -0.9 to -30.4 mV. Proteome disruption affected size and stability but not shape. Biosynthesized Ag-NPs using protein-free extracts ranged from 55 to 62 nm, and zeta potentials varied from -18 to -27 mV. Molecular docking studies and PASS results found no role for the metabolome in antibacterial activity. This suggests the antibacterial activity comes from Ag-NPs, not capping or reducing agents. Overall, the research affirmed the vital role of specific reducing metabolites in the biosynthesis of Ag-NPs, while proteins derived from biological extracts were found to solely affect their size and stability.

Reproductive strategy response of the fungi Sarocladium and the evaluation for remediation under stress of heavy metal Cd(II).

Cadmium (Cd) is documented as one of the most lethal metals and poses a major threat to all life forms in the environment due to its toxic effects. Bioremediation of hazardous metals has received considerable and growing interest over the years. The functional fungi with tolerance to the heavy metal Cd were screened from the mining soil samples. Two fungi isolates from coal mine soil were characterized as Sarocladium sp. M2 and Sarocladium sp. M6 based on morphological and partial ITS sequencing analysis. M2 and M6 exhibited high levels of resistance to cadmium, and they were investigated for their micro-morphology and application in heavy metal removal with different concentration Cd(II) (0, 50, 100, 150 and 200 mg/L). The colony morphology of M2 and M6 gradually become very similar to that of bacteria with the increase of cadmium concentration (150-200 mg/L). Micro-morphological studies showed that Cd(II) exposure caused the disappearance of conidial heads and the occurrence of hyphae breakage (100-200 mg/L Cd(II), which is consistent to the colony morphology results. The surface/volume ratio of the spores decreased with the presence of Cd(II). The removal potential of fungi for cadmium was quantified by atomic absorption spectrometry. M2 and M6 showed great potential as bioremediators for highly Cd(II)-contaminated environment. The highest Cd(II) biosorption capacity was 5.13 ± 0.21 mg/g for M2 and 6.04 ± 0.21 mg/g for M6. The highest heavy metal sorption by M2 removed 57.11% ± 4.45% Cd(II) while that of M6 removed 48.35% ± 1.44% Cd(II) in 200 mg/L initial concentration Cd(II). To the best of our knowledge, this is the first report that cadmium induced the change of reproduction mode of the Sarocladium, from conidia to arthrospores, which made the colony morphological modifications, from the fungi colony morphology to the bacteria colony morphology. The arthrospore-modified (hyphae breakage) seemed to accumulate greater amounts of heavy metals than filamentous hyphae formation.

Evaluation of copper-tolerant fungi isolated from Sarcheshmeh copper mine of Iran.

Mycoremediation, a subset of bioremediation, is considered an advanced method to eliminate environmental contaminations. To identify tolerant fungi to copper contamination and study the related gene expression, sampling was carried out from the soil of "Sarcheshmeh Copper Mine," which is one of the biggest open-cast copper mines in the world. A total of 71 fungal isolates were obtained and purified. Afterward, the inhibitory effect of different concentrations (1000, 1500, 3500, 4000, and 5500 ppm) of copper sulfate on mycelial growth was evaluated. Results indicated that only 5500 ppm of copper sulfate inhibited fungal growth compared to the control. Based on the bioassay experiments, three isolates including S3-1, S3-21, and S1-7, which were able to grow on solid and broth medium containing 5500 ppm of copper sulfate at different pH conditions, were selected and identified using molecular approaches. Also, laccase and metallothionein gene expression has been assessed in these isolates. According to the molecular identification using ITS1-5.8S- ITS2 region, isolates S3-1 and S1-7 were identified as Pleurotus eryngii, and isolate S3-21 belonged to the genus Sarocladium. In addition, P. eryngii showed laccase gene expression reduction after 8 days of exposure to copper sulfate. While in the genus Sarocladium, it increased (almost 2 times) from 6 to 8 days. Besides, metallothionein gene expression has increased from 6 to 8 days of copper sulfate treatment compared to the control which reveals its role in copper tolerance of all studied isolates. In this study, Pleurotus eryngii and Sarocladium sp. are introduced as heavy metal tolerant fungi and the related gene expression to copper tolerance was studied for the first time in Iran.

The genome and transcriptome of Sarocladium terricola provide insight into ergosterol biosynthesis.

Sarocladium terricola is a species of ascomycete fungus that has been recognized as a biocontrol agent for managing animal and plant pathogens, and exhibits significant potential as a feed additive. In this study, we utilized a combination of short-read Illumina sequencing and long-read PacBio sequencing to sequence, assemble, and analyze the genome of S. terricola. The resulting genome consisted of 11 scaffolds encompassing 30.27 Mb, with a GC content of 54.07%, and 10,326 predicted protein coding gene models. We utilized 268 single-copy ortholog genes to reconstruct the phylogenomic relationships among 26 ascomycetes, and found that S. terricola was closely related to two Acremonium species. We also determined that the ergosterol content of S. terricola was synthesized to nearly double levels when cultured in potato dextrose media compared to bean media (4509 mg/kg vs. 2382 mg/kg). Furthermore, transcriptome analyses of differentially expressed genes suggested that the ergosterol synthesis genes ERG3, ERG5, and ERG25 were significantly up-regulated in potato dextrose media. These results will help us to recognize metabolic pathway of ergosterol biosynthesis of S. terricloa comprehensivelly.

Cloning and sequence analysis of 4-O-α-d-isomaltooligosaccharylmaltooligosaccharide 1,4-α-isomaltooligosaccharohydrolase from Sarocladium kiliense U4520.

A novel enzyme, 4-O-α-d-isomaltooligosaccharylmaltooligosaccharide 1,4-α-isomaltooligosaccharohydrolase (IMM-4IH), was previously discovered from Sarocladium kiliense U4520. In order to identify the factors underlying the unique substrate specificity of IMM-4IH, we endeavored to determine the amino acid sequence of the enzyme. By comparing the partial amino acid sequence of the enzyme to whole genome sequencing data of S. kiliense U4520, the IMM-4IH gene was estimated. The putative gene was expressed in Pichia pastoris, and its activity and properties were found to be consistent with those of the native enzyme. Comparing the amino acid sequence of IMM-4IH with those in the CAZy database led to classification in the glycoside hydrolase family 49 (GH49). Several amino acids important for catalysis (Asp406, Asp425, and Asp426) and substrate recognition at subsites + 1 and -3 were estimated by multiple sequence alignment analysis. These results provide important information for characterizing IMM-4IH and other GH49 enzymes.

Sustainable production of biopesticides for common bean root rot control within the biorefinery approach: a Technology Readiness Level 3 experimental proof of concept.

Biopesticides are recognized as an efficient alternative to synthetic pesticides for pest and disease crop management. However, their commercial production processes use grains, generating large amounts of organic waste, even when agriculture waste or byproducts are the feedstock of choice. Frequently, these organic wastes are rich in nutrients that, after adequate treatment, can be used as nitrogen and carbohydrate sources for secondary metabolite production produced by microorganisms during submerged fermentation. In this sense, this study aimed to prove the concept that biopesticides could be produced under a full biorefinery process, using the entire biomass of an underexplored agroindustrial waste-damaged bean-as the main feedstock. A combination of sequential processes, including solid state fermentation, hydrolysis, and submerged fermentation, were designed for the production of two biopesticides (conventional-fungal conidia and second-generation secondary metabolite-cerulenin) from a high potential biological control agent strain Sarocladium oryzae BRM 59907. The combined processes, using damaged common bean grain as the main feedstock, provided biopesticides and organic fertilizer production that successfully controlled common bean root rot disease. This work proved to be possible the biopesticide production using a full biorefinery concept, inside the same productive chain, contributing to a sustainable environment and economy, together with animal and human health safety.

A colorimetric hydroxy naphthol blue based loop-mediated isothermal amplification detection assay targeting the ß-tubulin locus of Sarocladium oryzae infecting rice seed.

Sarocladium oryzae is a widely prevalent seed-borne pathogen of rice. The development of a rapid and on-site detection method for S. oryzae is therefore important to ensure the health of rice seeds. Loop-mediated isothermal amplification (LAMP) is ideal for field-level diagnosis since it offers quick, high-specific amplification of target template sequences at a single temperature. We designed primers based on the ß-tubulin region of S. oryzae. The LAMP technique devised was extremely sensitive, detecting the presence of the S. oryzae template at concentrations as low as 10 fg in 30 minutes at 65°C. The assay specificity was confirmed by performing the experiment with genomic DNA isolated from 22 different phytopathogens. Through the addition of hydroxy naphthol blue in the reaction process prior to amplification, a colour shift from violet to deep sky blue was seen in the vicinity of the target pathogen only. Finally, the LAMP assay was validated using live infected tissues, weeds and different varieties of seeds collected from different locations in Tamil Nadu, India. If developed into a detection kit, the LAMP assay developed in this study has potential applications in seed health laboratories, plant quarantine stations, and on-site diagnosis of S. oryzae in seeds and plants.

Colorimetric loop-mediated isothermal amplification assay for detection and ecological monitoring of Sarocladium oryzae, an important seed-borne pathogen of rice.

Accurate and timely disease detection plays a critical role in achieving sustainable crop protection. Globally, rice has been a staple crop for centuries plagued by the diseases that greatly hamper its productivity. Sheath rot, an emerging disease of rice caused by the seed-borne pathogen Sarocladium oryzae, has reportedly caused heavy losses to agricultural produce in recent years. Our study has led to the development and validation of a LAMP assay for early detection of S. oryzae, the causal agent of sheath rot from the live-infected tissues, seeds, weeds, and environmental samples. The assay could detect as low as 1.6 fg/µl of the pathogen in 15 min. The assay was implemented to bio-surveil the presence of this pathogen by testing it on three weed species (Echinochloa colona, Echinochloa crus-galli, and Cyperus teneriffae) growing around the rice fields. The results showed the presence of the pathogen in two of the weed species viz. E. colona and E. crus-galli. The assay was used to test 13 different rice varieties for the presence of S. oryzae in seeds. In total, three of the varieties did not show the presence of S. oryzae in their seeds while the rest were found to harbor the pathogen. The developed assay can effectively be used to detect and screen the presence of S. oryzae in live samples including seeds and field soil.

Identification of Genetic Diversity, Pyrrocidine-Producing Strains and Transmission Modes of Endophytic Sarocladium zeae Fungi from Zea Crops.

Genotyping by sequencing (GBS) was used to reveal the inherent genetic variation within the haploid fungi Sarocladium zeae isolated from diverse Zea germplasm, including modern Zea mays and its wild progenitors-the teosintes. In accordance with broad host relationship parameters, GBS analysis revealed significant host lineages of S. zeae genetic diversity, indicating that S. zeae genetic variation may associate with different evolutionary histories of host species or varieties. Based on a recently identified PKS-NRPS gene responsible for pyrrocidine biosynthesis in S. zeae fungi, a novel PCR assay was developed to discriminate pyrrocidine-producing S. zeae strains. This molecular method for screening bioactive strains of S. zeae is complementary to other approaches, such as chemical analyses. An eGFP-labelled S. zeae strain was also developed to investigate the endophytic transmission of S. zeae in Z. mays seedlings, which has further improved our understanding of the transmission modes of S. zeae endophytes in maize tissues.

Discovery of a novel glucanohydrolase, 4-α-isomaltooligosylglucose 4-glucanohydrolase, that can be used for efficient production of isomaltose.

We discovered a novel enzyme in our pursuit of an improved method for the production of isomaltose. The enzyme, 4-α-isomaltooligosylglucose 4-glucanohydrolase from Sarocladium kiliense U4520, recognizes the panose motif (α-d-Glcp-(1 â†’ 6)-α-d-Glcp-(1 â†’ 4)-d-Glcp) and hydrolyzes the α-1,4-glucosidic bond on the reducing end side with respect to the α-1,6-glucosidic bond. The structure on the non-reducing end of the panose motif is important for the recognition of the substrate by the enzyme, and the substrate specificity is unique and distinguished from previously reported enzymes. The enzyme catalyzes the hydrolysis of panose with a kcat/Km of 31.2 s-1mM-1, and catalysis results in anomeric inversion. These enzymatic properties suggest that this enzyme will pair well with 1,4-α-glucan 6-α-glucosyltransferase from Bacillus globisporus N75 in the efficient production of isomaltose from starch.

Potassium Deficiency in Rice Aggravates Sarocladium oryzae Infection and Ultimately Leads to Alterations in Endophyte Communities and Suppression of Nutrient Uptake.

Sheath rot disease is an emerging fungal disease in rice, whose infection causes severe yield loss. Sarocladium oryzae (S. oryzae) is the major causal agent. Previous study has demonstrated that rice deficiency in potassium (K) aggravates S. oryzae infection. However, the effects of S. oryzae infection on the nutrient-uptake process, endophyte communities, and hormone level of host plant under K-deficiency condition remain unclear, the mechanism of K mediated S. oryzae infection needs to be further study. The present study analyzed alterations in the endophytic community and nutrient-uptake process of host plants through an exogenous inoculation of S. oryzae in pot and hydroponics experiments. S. oryzae infection sharply increased the relative abundance of Ascomycota and decreased the Shannon and Simpson index of the endophytic community. Compared with the K-sufficient rice infected with S. oryzae, K-starved rice infected with S. oryzae (-K + I) increased the relative abundance of Ascomycota in leaf sheaths by 52.3%. Likewise, the -K + I treatment significantly decreased the Shannon and Simpson indexes by 27.7 and 25.0%, respectively. Sufficient K supply increased the relative abundance of Pseudomonas spp. in the host plant. S. oryzae infection profoundly inhibited the nutrient uptake of the host plant. The accumulation of oleic acid and linoleic acid in diseased rice decreased the biosynthesis of jasmonic acid (JA), and the content of JA was lowest in the -K + I treatment, which suppressed K+ uptake. These results emphasize the importance of K in resistance to S. oryzae infection by modulating endophyte community diversity and enhancing the nutrient-uptake capacity of the host plant.

First Report of Fusarium Sheath Rot of Rice Caused by Fusarium incarnatum-equiseti Species Complex in the United States.

Fungal diseases, including sheath rot (Sarocladium oryzae), cause significant losses of yield and milling quality of rice (Oryza sativa). In August 2021, symptoms like sheath rot were observed on 20% of rice plants (cv. Presidio) in 1-hectare field in Eagle Lake, Texas. Initial lesions occurred on the upper flag leaf sheaths and were oblong or irregular oval, with gray to light brown centers, and a dark reddish-brown diffuse margin. Lesions enlarged, coalesced, and covered a large area of the sheath. Infection led to panicle rot with kernels turning dark brown. Unlike sheath rot, sheath infection also led to inside culm infection with irregular dark brown lesions. Infected tissue pieces were sterilized with 1% NaOCl for 2 min, followed by 75% ethanol for 30 s, washed in sterile H2O three times, air dried and incubated on PDA at 27℃. Fungal isolates were obtained from 15 diseased plant samples and their singled-spored fungal colonies were whitish, loosely floccose and produced light yellow pigmentation. On carnation leaf agar, macroconidia were slightly curved and tapered at the ends, with 3 to 5 septa, and measured 17.5 to 34.3 × 3.1 to 5.0 µm. Microconidia were ovoid, usually with 0 to 1 septum and were 4.0 to 15.5 × 2.5 to 4.5 µm. Spherical shaped chlamydospores were produced in chain. These morphological characteristics were consistent to those described for Fusarium incarnatum-equiseti species complex (O'Donnell et al. 2009), including F. incarnatum (Wang et al. 2021) and F. equiseti (Avila et al. 2019). For molecular identification, DNA of a representative isolate was extracted and ITS, LSU, and EF1 of the fungus were amplified using the primers of ITS1/ITS4 (Wang et al. 2014), D1/D2 domain region of LSU (Fell et al. 2000), and EF1 (Wang et al. 2014), respectively, and sequenced. The ITS sequence (OL344049) was 99.61% identical to F. incarnatum-equiseti species complex (FD_01692) in Fusarium-ID database and 99.61% identical to F. equiseti (LC514690, KY523100, MW016539) and F. incarnatum (MH979697) in NCBI database. The LSU sequence (OK559512) was 98.77% similar to F. equiseti (MN877913, MN368509) and F. incarnatum (MH877332, MH877326); the EF1 sequence (OK570044) was 99.27% similar to F. equiseti (MK278902) in NCBI database. A phylogenetic analysis based on the concatenated nucleotide sequences grouped this isolate in the F. incarnatum-equiseti species complex clade at 100% bootstrap support. To evaluate pathogenicity, a conidial suspension of 1 x 106 conidia/ml or sterilized water (the controls) was injected into the sheaths and young panicles of three rice plants (cv. Presidio) at boot. Treated plants were maintained in a greenhouse at 25 to 30℃. After 3 weeks, typical symptoms, like those observed in the field, developed on the inoculated plants but not on the controls. The same fungus was consistently re-isolated from the diseased plants. To our knowledge, this is the first report of Fusarium sheath rot caused by F. incarnatum-equiseti species complex in rice in the U. S. F. incarnatum-equiseti species complex has been reported to be associated with panicle infection in wild rice (O. latifolia) in Brazil (Tralamazza et al. 2021). F. incarnatum has also been reported to cause panicle rot in China (Wang et al. 2021). F. proliferatum has been reported to cause Fusarium sheath rot in India (Prabhukarthikeyan et al. 2021) and the U. S. (Cartwright et al. 1995). This research demonstrates the potential of different pathogens being involved in causing sheath rot of rice.

A sensitive, accurate, and high-throughput gluco-oligosaccharide oxidase-based HRP colorimetric method for assaying lytic polysaccharide monooxygenase activity.

BACKGROUND: The AA9 (auxiliary activities) family of lytic polysaccharide monooxygenases (AA9 LPMOs) is a ubiquitous and diverse group of enzymes in the fungal kingdom. They catalyse the oxidative cleavage of glycosidic bonds in lignocellulose and exhibit great potential for biorefinery applications. Robust, high-throughput and direct methods for assaying AA9 LPMO activity, which are prerequisites for screening LPMOs with excellent properties, are still lacking. Here, we present a gluco-oligosaccharide oxidase (GOOX)-based horseradish peroxidase (HRP) colorimetric method for assaying AA9 LPMO activity. RESULTS: We cloned and expressed a GOOX gene from Sarocladium strictum in Trichoderma reesei, purified the recombinant SsGOOX, validated its properties, and developed an SsGOOX-based HRP colorimetric method for assaying cellobiose concentrations. Then, we expressed two AA9 LPMOs from Thielavia terrestris, TtAA9F and TtAA9G, in T. reesei, purified the recombinant proteins, and analysed their product profiles and regioselectivity towards phosphoric acid swollen cellulose (PASC). TtAA9F was characterized as a C1-type (class 1) LPMO, while TtAA9G was characterized as a C4-type (class 2) LPMO. Finally, the SsGOOX-based HRP colorimetric method was used to quantify the total concentration of reducing lytic products from the LPMO reaction, and the activities of both the C1- and C4-type LPMOs were analysed. These LPMOs could be effectively analysed with limits of detection (LoDs) less than 30 nmol/L, and standard curves between the A515 and LPMO concentrations with determination coefficients greater than 0.994 were obtained. CONCLUSIONS: A novel, sensitive and accurate assay method that directly targets the main activity of both C1- and C4-type AA9 LPMOs was established. This method is easy to use and could be performed on a microtiter plate for high-throughput screening of AA9 LPMOs with desirable properties.

Biocontrol Ability and Production of Volatile Organic Compounds as a Potential Mechanism of Action of Olive Endophytes against Colletotrichum acutatum.

Olive anthracnose, mainly caused by Colletotrichum acutatum, is considered a key biotic constraint of the olive crop worldwide. This work aimed to evaluate the ability of the endophytes Aureobasidium pullulans and Sarocladium summerbellii isolated from olive trees to reduce C. acutatum growth and anthracnose symptoms, and to assess A. pullulans-mediated changes in olive fruit volatile organic compounds (VOCs) and their consequences on anthracnose development. Among the endophytes tested, only A. pullulans significantly reduced the incidence (up to 10-fold) and severity (up to 35-fold) of anthracnose in detached fruits, as well as the growth (up to 1.3-fold), sporulation (up to 5.9-fold) and germination (up to 3.5-fold) of C. acutatum in dual culture assays. Gas chromatography-mass spectrometry analysis of olives inoculated with A. pullulans + C. acutatum and controls (olives inoculated with C. acutatum, A. pullulans or Tween) led to the identification of 37 VOCs, with alcohols being the most diversified and abundant class. The volatile profile of A. pullulans + C. acutatum revealed qualitative and quantitative differences from the controls and varied over the time course of microbial interactions. The most significant differences among treatments were observed at a maximal reduction in anthracnose development. At this stage, a set of VOCs, particularly Z-3-hexen-1-ol, benzyl alcohol and nonanal, were highly positively correlated with the A. pullulans + C. acutatum treatment, suggesting they play a critical role in anthracnose reduction. 6-Methyl-5-hepten-2-one and 2-nonanone were positively associated with the C. acutatum treatment and thus likely have a role in pathogen infection.

Non-valvular Infective Endocarditis Caused by Sarocladium kiliense in an Immunocompromised Patient with Aplastic Anemia.

Sarocladium kiliense is ubiquitous in the human environment and is an emerging opportunistic pathogen, especially among immunocompromised hosts. A 77-year-old man diagnosed with aplastic anemia suffered from non-valvular endocarditis. After he passed away, fungal hyphae were observed in several lesions on a postmortem examination. Polymerase chain reaction (PCR) and a DNA sequence analysis revealed S. kiliense as the causative organism. This is the first case report of non-valvular fungal endocarditis caused by S. kiliense identified by PCR and a DNA sequence analysis in an immunocompromised patient. Although rare, invasive fungal infection caused by S. kiliense should be considered in immunocompromised hosts.

Case Report: A Case of Sarocladium strictum Meningoencephalitis in an Immunocompetent Patient After Invasive Operation.

As an opportunistic phytopathogen, Sarocladium strictum has only been shown to cause neurological disease in immunocompromised patients, where antifungal therapy was not effective. We report a case of Sarocladium strictum meningoencephalitis in an apparently immunocompetent young woman who presented with severe headache and slight fever after undergoing transnasal endoscopic repair of cerebrospinal fluid rhinorrhea. Chronic sinusitis and suspicious intracranial fungal lesions were observed on enhanced magnetic resonance imaging (MRI). Both culture and metagenomic next-generation sequencing of her cerebrospinal fluid were positive for Sarocladium strictum. After local debridement, treatment with amphotericin B plus voriconazole and Ommaya reservoir implantation, the patient improved significantly. Unfortunately, her symptoms worsened again despite plenty of antifungal therapy for a month.

Fatal disseminated infection due to Sarocladium kiliense in a diabetic patient with COVID-19.

Sarocladium kiliense is a soil saprophytic mold with worldwide distribution, which can infect humans and other mammals, sporadically. The clinical manifestations include mycetoma, onychomycosis, keratomycosis, pneumonia, and arthritis. Here, we present a disseminated infection due to S. kiliense in a diabetic patient infected to coronavirus disease 2019 (COVID-19) from Isfahan, Iran.