Dimorphic Fungus Research Articles

Puzzling paracoccidioidomycosis: Factors associated with the severity of Paracoccidioides lutzii infections

ObjectivesHistorically, the Brazilian Central-West region has had high numbers of paracoccidioidomycosis (PCM) cases caused by the dimorphic fungus Paracoccidioides lutzii. MethodsThis epidemiological, observational, analytical, cross-sectional study was performed to investigate the clinical and laboratory data of 44 PCM patients with a culture-proven P. lutzii infection. All patients were referred to the Systemic Mycosis Center, Júlio Muller University Hospital, Cuiabá, Brazil, during January 2017 to March 2020. The neutrophil to lymphocyte ratio (NLR) was calculated and dichotomized by its median value to include in the identification of factors associated with severity. ResultsAt admission, 13 (31.7%) patients showed the disseminated multifocal chronic form of PCM and 16 (36.4%) patients met the clinical severity criteria. Treatment prescribed on admission did not follow the recommendations of the Brazilian Guideline for the Clinical Management of Paracoccidioidomycosis in 26% of the severe PCM cases (prevalence ratio 0.26, 95% confidence interval 0.14–0.49; P < 0.0001). Patients with severe PCM had a higher NLR that was greater than the median (≥4.11). ConclusionsThe NLR biomarker complements the criteria for PCM severity. Applying the low-cost NLR test can greatly increase the diagnostic sensitivity when screening patients for PCM and contribute to better control of the disease, management of complications, and therapeutic strategies.

The Known Unknowns of the Immune Response to Coccidioides.

Coccidioidomycosis, otherwise known as Valley Fever, is caused by the dimorphic fungi Coccidioides immitis and C. posadasii. While most clinical cases present with self-limiting pulmonary infection, dissemination of Coccidioides spp. results in prolonged treatment and portends higher mortality rates. While the structure, genome, and niches for Coccidioides have provided some insight into the pathogenesis of disease, the underlying immunological mechanisms of clearance or inability to contain the infection in the lung are poorly understood. This review focuses on the known innate and adaptive immune responses to Coccidioides and highlights three important areas of uncertainty and potential approaches to address them. Closing these gaps in knowledge may enable new preventative and therapeutic strategies to be pursued.

Diagnosis of Pulmonary Infections Due to Endemic Fungi.

Endemic mycoses including Histoplasma, Blastomyces, Coccidioides, Paracoccidioides, and Talaromyces are dimorphic fungi that can cause a variety of clinical manifestations, including respiratory infections. Their pulmonary presentations are variable, and diagnosis is often delayed as they can mimic other infectious and non-infectious causes of pulmonary disease. Delay in diagnosis can lead to unnecessary antibiotic use, repeat hospitalizations, and increased morbidity and mortality. The diagnosis of endemic fungal pulmonary infections often relies on multiple diagnostic tests including culture, tissue histopathology, antigen assays, and antibody assays. Due to the increased use of immunosuppressive agents and the widening geographic ranges where these infections are being found, the prevalence of endemic fungal infections is increasing. Physicians need to be aware of the clinical manifestations of pulmonary infections due to endemic fungal in order to ensure that the proper diagnostic work up is obtained promptly. A high index of suspicion is particularly important in patients with suspected pulmonary infections who have failed to improve despite antibiotics in the appropriate setting. We present a review diagnostic testing for pulmonary infections due to endemic mycoses.

Transcriptional Analysis of Coccidioides immitis Mycelia and Spherules by RNA Sequencing.

Coccidioides immitis and C. posadasii are dimorphic fungi that transform from mycelia with internal arthroconidia in the soil to a tissue form known as a spherule in mammals. This process can be recapitulated in vitro by increasing the temperature, CO2 and changing other culture conditions. In this study, we have analyzed changes in gene expression in mycelia and young and mature spherules. Genes that were highly upregulated in young spherules include a spherule surface protein and iron and copper membrane transporters. Genes that are unique to Coccidioides spp. are also overrepresented in this group, suggesting that they may be important for spherule differentiation. Enriched GO terms in young spherule upregulated genes include oxidation-reduction, response to stress and membrane proteins. Downregulated genes are enriched for transcription factors, especially helix–loop–helix and C2H2 type zinc finger domain-containing proteins, which is consistent with the dramatic change in transcriptional profile. Almost all genes that are upregulated in young spherules remain upregulated in mature spherules, but a small number of genes are differentially expressed in those two stages of spherule development. Mature spherules express more Hsp31 and amylase and less tyrosinase than young spherules. Some expression of transposons was detected and most of the differentially expressed transposons were upregulated in spherules.

An inexpensive point-of-care immunochromatographic test for Talaromyces marneffei infection based on the yeast phase specific monoclonal antibody 4D1 and Galanthus nivalis agglutinin.

Talaromyces marneffei is a thermally dimorphic fungus that causes opportunistic systemic mycoses in patients with AIDS or other immunodeficiency syndromes. The purpose of this study was to develop an immunochromatographic strip test (ICT) based on a solid phase sandwich format immunoassay for the detection of T. marneffei antigens in clinical urine specimens. The T. marneffei yeast phase specific monoclonal antibody 4D1 (MAb4D1) conjugated with colloidal gold nanoparticle was used as a specific signal reporter. Galanthus nivalis Agglutinin (GNA) was adsorbed onto nitrocellulose membrane to serve as the test line. Similarly, a control line was created above the test line by immobilization of rabbit anti-mouse IgG. The immobilized GNA served as capturing molecule and as non-immune mediated anti-terminal mannose of T. marneffei antigenic mannoprotein. The MAb4D1–GNA based ICT showed specific binding activity with yeast phase antigen of T. marneffei, and it did not react with other common pathogenic fungal antigens. The limit of detection of this ICT for T. marneffei antigen spiked in normal urine was approximately 0.6 μg/ml. The diagnostic performance of the ICT was validated using 341 urine samples from patents with culture- confirmed T. marneffei infection and from a control group of healthy individuals and patients with other infections in an endemic area. The ICT exhibited 89.47% sensitivity, 100% specificity, and 97.65% accuracy. Our results demonstrate that the urine-based GNA–MAb4D1 based ICT produces a visual result within 30 minutes and that the test is highly specific for the diagnosis of T. marneffei infection. The findings validate the deployment of the ICT for clinical use.

Metallothioneins reprograms T cell response towards pulmonary Histoplasma capsulatum through altering their metabolic profile

Abstract Histoplasma capsulatum (Hc) is a dimorphic fungus that is affected by environmental temperature. Hc is distributed worldwide. When inhaled, the microconidia and spores reach the lung and are converted to the pathogenic yeast phase within the phagosomes of macrophages. The Hc replicates intracellularly until these phagocytes are activated by cytokines, typically from activated T cells. Excluding iron, the effect of trace metals on Hc interaction with innate immune cells has been inadequately studied. The cells’ metabolic processes are altered, and the cytokine macro/microenvironment change in the pathogen’s face. Zn2+ metalloproteins constitute about 10% of the mammalian proteome, and many transcription factors and enzymes use this metal. Metalloproteins are linked to many energy-producing pathways, including carbohydrate metabolism, amino acid metabolism, and the redox chain. Our experiments showed that ablation of Metallothioneins 1 and 2 (MT1/2) decreases the immune system’s ability to clear Hc from the lungs. The numbers of T cells are not affected by the MT1/2 ablation; rather, the cells’ activity is decreased. MT1/2 ablated dendritic cells (DC) are not defective. MT1/2 ablated CD4 T cells show an alternative metabolic profile. MT1/2 ablated CD4 T cells show a decrease in their genome acetylation.

Disseminated coccidioidomycosis in a patient with juvenile idiopathic arthritis receiving infliximab

BackgroundCoccidioides immitis is a dimorphic fungus endemic to the arid climates of the Southwest United States, Mexico and parts of Central and South America. Human infection occurs through inhalation of spores with less than half of exposures progressing to a symptomatic state that primarily consists of pulmonary manifestations. Disseminated coccidioidomycosis is exceedingly rare, occurring in fewer than 1 % of symptomatic infections. Through hematogenous spread, the fungus can infect most organ systems and may be fatal without systemic antifungal treatment. Individuals with impaired cell-mediated immunity either from primary immunodeficiency disorders or secondary to immunosuppression with medications such as tumor necrosis factor alpha (TNF-α) inhibitors have increased risk of disseminated coccidioidomycosis and previous cases of coccidioidomycosis have been reported with biologic therapy.Case presentationWe present a case of disseminated coccidioidomycosis in a 16-year-old female with polyarticular juvenile idiopathic arthritis (JIA) being treated with prednisone, methotrexate, and infliximab. The patient presented with symptoms of meningeal irritation, bilateral choroidal lesions, and necrotizing peripheral pneumonia. Her infection was thought to be a reactivation of coccidioidomycosis given her history of resolved pneumonia that occurred after traveling to Arizona, New Mexico, and El Paso one year prior to presentation. Following diagnosis, she improved with discontinuation of her immunosuppressive medications and two weeks of intravenous amphotericin B and fluconazole with plans for lifetime treatment with fluconazole while immunosuppressed. Due to worsening arthritis, she will begin tofacitinib and continue close monitoring of chest x-rays and coccidioides antibody.ConclusionsPatients undergoing immunosuppressive therapy for rheumatological conditions are at increased risk of disseminated coccidioidomycosis and should be evaluated with high suspicion when presenting with atypical symptoms and history of travel to endemic regions.

Life Cycle Dominates the Volatilome Character of Dimorphic Fungus Coccidioides spp.

ABSTRACTValley fever (coccidioidomycosis) is an endemic fungal pneumonia of the North and South American deserts. The causative agents of Valley fever are the dimorphic fungi Coccidioides immitis and C. posadasii, which grow as mycelia in the environment and as spherules within the lungs of vulnerable hosts. Current diagnostics for Valley fever are severely lacking due to poor sensitivity and invasiveness, contributing to a 23-day median time to diagnosis, and therefore, new diagnostic tools are needed. We are working toward the development of a breath-based diagnostic for coccidioidomycosis, and in this initial study, we characterized the volatile metabolomes (or volatilomes) of in vitro cultures of Coccidioides. Using solid-phase microextraction (SPME) and comprehensive two-dimensional gas chromatography coupled to time of flight mass spectrometry (GC×GC-TOFMS), we characterized the volatile organic compounds (VOCs) produced by six strains of each species during mycelial or spherule growth. We detected a total of 353 VOCs that were at least 2-fold more abundant in a Coccidioides culture than in medium controls and found that the volatile metabolome of Coccidioides is more dependent on the growth phase (spherules versus mycelia) than on the species. The volatile profiles of C. immitis and C. posadasii have strong similarities, indicating that a single suite of Valley fever breath biomarkers can be developed to detect both species.IMPORTANCE Coccidioidomycosis, or Valley fever, causes up to 30% of community-acquired pneumonias in highly populated areas of the U.S. desert southwest where the disease is endemic. The infection is difficult to diagnose by standard serological and histopathological methods, which delays appropriate treatment. Therefore, we are working toward the development of breath-based diagnostics for Valley fever. In this study, we characterized the volatile metabolomes (or volatilomes) of six strains each of Coccidioides immitis and C. posadasii, the dimorphic fungal species that cause Valley fever. By analyzing the volatilomes during the two modes of growth of the fungus—mycelia and spherules—we observed that the life cycle plays a significant role in the volatiles produced by Coccidioides. In contrast, we observed no significant differences in the C. immitis versus C. posadasii volatilomes. These data suggest that life cycle, rather than species, should guide the selection of putative biomarkers for a Valley fever breath test.

Quantification of oxygen transfer coefficients in simulated hydrocarbon-based bioprocesses in a bubble column bioreactor.

This study investigates the overall volumetric oxygen transfer coefficient (KLa) in multiphase hydrocarbon-based bioprocess under a range of hydrocarbon concentrations (HC), solid loadings (deactivated yeast) (SL) and superficial gas velocities (UG) in a bubble column reactor (BCR). KLa increased with increasing UG in the air-water system; due to an increase in the number of small bubbles which enhanced gas holdup. In air-water-yeast systems, the initial addition of yeast increased KLa significantly. Further increases in SL reduced KLa, due to increases in the bubble size with increasing SL. KLa decreased when HC was added in air-water-hydrocarbon systems. However, UG, SL and HC affected KLa differently in air-water-yeast-hydrocarbon systems: an indication of the complex interactions between the yeast and hydrocarbon phases which changed the system's hydrodynamics and therefore affected KL. This work illustrates the effect of the operating conditions (SL, HC and UG) on oxygen transfer behaviour in multiphase systems.

Emergomycosis, an Emerging Systemic Mycosis in Immunocompromised Patients: Current Trends and Future Prospects.

Recently, the global emergence of emergomycosis, a systemic fungal infection caused by a novel dimorphic fungus Emergomyces species has been observed among immunocompromised individuals. Though initially classified under the genus Emmonsia, a taxonomic revision in 2017 based on DNA sequence analyses placed five Emmonsia-like fungi under a separate genus Emergomyces. These include Emergomyces pasteurianus, Emergomyces africanus, Emergomyces canadensis, Emergomyces orientalis, and Emergomyces europaeus. Emmonsia parva was renamed as Blastomyces parvus, while Emmonsia crescens and Emmonsia sola remained within the genus Emmonsia until a taxonomic revision in 2020 placed both the species under the genus Emergomyces. However, unlike other members of the genus, Emergomyces crescens and Emergomyces sola do not cause disseminated disease. The former causes adiaspiromycosis, a granulomatous pulmonary disease, while the latter has not been associated with human disease. So far, emergomycosis has been mapped across four continents: Asia, Europe, Africa and North America. However, considering the increasing prevalence of HIV/AIDS, it is presumed that the disease must have a worldwide distribution with many cases going undetected. Diagnosis of emergomycosis remains challenging. It should be considered in the differential diagnosis of histoplasmosis as there is considerable clinical and histopathological overlap between the two entities. Sequencing the internal transcribed spacer region of ribosomal DNA is considered as the gold standard for identification, but its application is compromised in resource limited settings. Serological tests are non-specific and demonstrate cross-reactivity with Histoplasma galactomannan antigen. Therefore, an affordable, accessible, and reliable diagnostic test is the need of the hour to enable its diagnosis in endemic regions and also for epidemiological surveillance. Currently, there are no consensus guidelines for the treatment of emergomycosis. The recommended regimen consists of amphotericin B (deoxycholate or liposomal formulation) for 1–2 weeks, followed by oral itraconazole for at least 12 months. This review elaborates the taxonomic, clinical, diagnostic, and therapeutic aspects of emergomycosis. It also enumerates several novel antifungal drugs which might hold promise in the treatment of this condition and therefore, can be potential areas of future studies.

009 Hyperthermia controls DAB2 transcription in macrophage through inducing the separation of cJun and cFos heterodimers

Sporotrichosis is an emergent subcutaneous mycosis of humans and some animals caused by dimorphic fungi of the Sporothrix schenckii. Hyperthermia can effectively treat sporotrichosis by regulating the phenotypic regulation of macrophages which is critical for controlling tissue inflammation and resolution. The adaptor protein disabled homolog 2 (DAB2), a regulator of phenotypic polarization in macrophages, have been identified to inhibit an inflammatory phenotype of the macrophages. However, whether and how hyperthermia act on the immune regulation of macrophages through DAB2 needs more research and complement. In this study, mouse bone marrow extracted primary macrophages (BMDMs) and cell line ANA-1 were used to investigate the regulation of DAB2 gene transcription by hyperthermia in vitro. Immunofluorescence was used to examine sub-localization of AP-1 complex within the cells under the 42°C conditions. Chromatin immunoprecipitation was used to detect whether 42°C stimulation affect the binding of AP-1complex to DAB2 gene. Conditional DAB2 knockout mice were used to evaluate the role of DAB2 in Sporotrichosis. Our results show that 42°C stimulation downregulated the expression of c-Jun and c-Fos, and led to the separation of c-Jun and c-Fos dimers, causing the downregulated transcription of DAB2. In conclusion, topical hyperthermia treatment can inhibit the transcription of DAB2 gene, promote macrophage M1 polarization, and promote the treatment of sporotrichosis via AP-1 complex.

A global view on fungal infections in humans and animals: infections caused by dimorphic fungi and dermatophytoses.

Fungal infections are still underappreciated and their prevalence is underestimated, which renders them a serious public health problem. Realistic discussions about their distribution, symptoms, and control can improve management and diagnosis and contribute to refinement of preventive actions using currently available tools. This article represents an overview of dermatophytes and endemic fungi that cause infections in humans and animals. In addition, the impact of climate change on the fungal spread is discussed. The endemic fungal infections characterized in this article include coccidioidomycosis, histoplasmosis, blastomycosis, lobomycosis, emergomycosis and sporotrichosis. Moreover the geographic distribution of these fungi, which are known to be climate sensitive and/or limited to endemic tropical and subtropical areas, is highlighted. In turn, dermatophytes cause superficial fungal infections of skin, hairs and nails, which are the most prevalent mycoses worldwide with a high economic burden. Therefore, the possibility of causing zoonoses and reverse zoonoses by dermatophytes is highly important. In conclusion, the article illustrates the current issues of the epidemiology and distribution of fungal diseases, emphasizing the lack of public programmes for prevention and control of these types of infection.

High prevalence and mortality due to Histoplasma capsulatum in the Brazilian Amazon: An autopsy study.

BackgroundHistoplasmosis is acquired by inhalation of spores of the dimorphic fungus Histoplasma spp. Although this pathogen is distributed worldwide, it is more prevalent in the Americas. However, the real burden of histoplasmosis remains undefined in many endemic regions.MethodologyWe conducted a series of 61 autopsies to individuals who died in a hospital in the Brazilian Amazon focused on infectious diseases. We performed a detailed histological and microbiological evaluation with genetic characterization of Histoplasma strains with the aim to evaluate the contribution of histoplasmosis to morbidity and mortality. Additionally, we assessed the clinicopathological correlation.Principal findingsEvidence of Histoplasma infection was detected in 21 patients (34%). Eight cases were disseminated infections, all of them occurred in HIV-positive patients. Six cases were localized histoplasmosis, limited to the lungs. In seven patients Histoplasma DNA was detected by PCR in patients with no histological lesions. Histoplasma infection was detected in 38% of HIV-positive patients and was a major contributor to death in 22% of them. Lungs, liver and spleen were affected in all cases of disseminated histoplasmosis. Phylogenetic analysis of the strains suggested a high diversity of Histoplasma species circulating in the Brazilian Amazon. Histoplasmosis was clinically missed in 75% of the disseminated infections.ConclusionsThe high incidence of histoplasmosis, the low index of clinical suspicion, and the severity of the disseminated disease highlight the need of proactively implementing sensitive routine screening methods for this pathogen in endemic areas. Antifungal prophylaxis against Histoplasma should be encouraged in the severely immunocompromised HIV patients in these areas. In conclusion, substantial mortality is associated with disseminated histoplasmosis among HIV-positive patients in the Brazilian Amazon.

Adenosine Triphosphate Released by Candida albicans Is Associated with Reduced Skin Infectivity

Skin is a barrier tissue that is exposed to both commensal microorganisms and pathogens. Candida albicans is a dimorphic fungus that typically grows as a commensal at barrier surfaces but can also become pathogenic (Gow et al., 2011). Pathogenic cutaneous C. albicans infection induces type-17 immunity that provides host defense (Kashem and Kaplan, 2016). The innate response of C. albicans skin infections is triggered by the recognition of the pathogen-derived toll-like receptor and Dectin-1 ligands (Netea et al., 2008).

Paracoccidioidomycosis in the northern region of Rio Grande do Sul

Background and Objectives: Paracoccidioidomycosis (PCM) is a systemic disease caused by the dimorphic fungus Paracoccidioides brasiliensis found in the tropical and subtropical regions of Latin America. This study aimed to perform a retrospective analysis of PCM cases from the northern region of Rio Grande do Sul, Brazil. Methods: A total of 200 records of PCM cases from 1995 to 2015 were analyzed, diagnosed at the local reference pathology laboratory. Results: Of the patients, 185 were male and 15 female. Patients ranged in age from 31 to 80 years, the largest proportion (35,5%) being aged between 51 and 60 years and living or working in the countryside. Clinical samples were mostly obtained from the oral cavity, followed by the oropharynx, lungs, brain, skin, and prostate. Conclusion: PCM is endemic in the south of Brazil, as the local economy is largely based on agricultural activities, favoring the contact of the population with P. brasiliensis. Due to the great similarity between PCM symptoms and other respiratory diseases, the differential diagnosis is essential for the correct treatment of the disease and to avoid its progression.

Antifungal Susceptibility Profiles of Olorofim (Formerly F901318) and Currently Available Systemic Antifungals against Mold and Yeast Phases of Talaromyces marneffei.

In vitro antifungal susceptibility profiling of 32 clinical and environmental Talaromyces marneffei isolates recovered from southern China was performed against olorofim and 7 other systemic antifungals, including amphotericin B, 5-flucytosine, posaconazole, voriconazole, caspofungin, and terbinafine, using CLSI methodology. In comparison, olorofim was the most active antifungal agent against both mold and yeast phases of all tested Talaromyces marneffei isolates, exhibiting an MIC range, MIC50, and MIC90 of 0.0005 to 0.002 μg/ml, 0.0005 μg/ml, and 0.0005 μg/ml, respectively.

Nasopharyngeal histoplasmosis– A rare case report

Histoplasmosis is a dimorphic fungus which is widely distributed among western countries like America and in some parts of the tropical region. It is acquired by inhaling microconidia. Depending on the immune status patient may develop acute or chronic pulmonary diseases or disseminated infection. The disseminated form of nasopharyngeal region can present as ulceration, rhino sinusitis, nasal obstruction, dyspnoea, hoarseness of voice and clinically it can mimic carcinoma. Microscopically, the differential diagnosis of histoplasmosis includes Leishmaniasis, Cryptococci neoformans, Candida glabrata, Pneumocystis jiroveci and Penicillium marneffei infections. The diagnosis of histoplasma is dependent on histopathological examination and demonstration by special stains, where the latter serves the gold standard in demonstrating organism on light microscopy. Keywords: Fungal infection, Histoplasmosis, Nasopharyngeal lesion, Paediatric age.

A single-cell analysis of the Arabidopsis vegetative shoot apex.

The shoot apical meristem allows for reiterative formation of new aerial structures throughout the life cycle of a plant. We use single-cell RNA sequencing to define the cellular taxonomy of the Arabidopsis vegetative shoot apex at the transcriptome level. We find that the shoot apex is composed of highly heterogeneous cells, which can be partitioned into 7 broad populations with 23 transcriptionally distinct cell clusters. We delineate cell-cycle continuums and developmental trajectories of epidermal cells, vascular tissue, and leaf mesophyll cells and infer transcription factors and gene expression signatures associated with cell fate decisions. Integrative analysis of shoot and root apical cell populations further reveals common and distinct features of epidermal and vascular tissues. Our results, thus, offer a valuable resource for investigating the basic principles underlying cell division and differentiation in plants at single-cell resolution.

Development of CRISPR-Cas9 genome editing system in Talaromyces marneffei

Talaromyces marneffei is an important pathogenic thermally dimorphic fungus causing systemic talaromycosis mainly prevalent in Southeast Asia. The dimorphic transition between mycelium and yeast is considered crucial for the pathogenicity of T. marneffei. However, the lack of genetic toolbox has been a major impediment for understanding its pathogenicity. Here a CRISPR-Cas9 system was developed to facilitate genetic manipulations in this organism. In this study, the CRISPR-Cas9 gene editing system uses a native U6 snRNA promoter from T. marneffei to drive the expression of sgRNA. Employing this system and PEG-mediated protoplast transformation, the sakA gene was mutated. Sanger sequencing confirmed nearly 40% site-directed mutation rate. The phenotype analysis confirmed the sakA gene function in T. marneffei dimorphic transition. Our study provided a powerful genome-manipulating tool, which could accelerate studies on T. marneffei for further revealing the mechanisms of its pathogenicity.

Antifungal Properties of Protein Extracts from Thai Medicinal Plants to Opportunistic Fungal Pathogens

Antifungal proteins or peptides (AFPs) are the natural products produced by several life forms including plants as the first-line defenses to infections. The AFPs from Thai plants can be used as new therapeutic agents in the world with growing resistance to conventional antifungals. This study determined the antifungal activities of protein extracts from Thai medicinal plants against important human opportunistic fungi, Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, and Talaromyces marneffei. Total crude protein supernatants and their precipitated proteins from 10 Thai medicinal plants with the historical usage for treatment of fungal infection were prepared. Most of the protein extracts showed antifungal activities to the tested fungi. The most effective reactivity found in the extracts from Rhinacanthus nasutus, Andrographis paniculata, and Psidium guajava by showing highest activity to T. marneffei ATCC200051 (yeast phase), followed by C. neoformans ATCC90112, C. albicans ATCC90028, T. marneffei ATCC200051 (mold phase), and A. fumigatus NCPF7367. The precipitated proteins from R. nasutus and A. paniculata containing antifungal properties were selected for partial purification by size cut-off membrane centrifugation and tested for antifungal activities. A colorimetric broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) to anti-T. marneffei. The partially purified fractions from A. paniculata, and R. nasutus showed anti-T. marneffei activity with the MIC and MFC values ranged from 2 to 128 μg/mL and 16 to &gt;128 μg/mL, respectively. Therefore, A. paniculata and R. nasutus can be further subjected to the study of the therapeutic antifungals.