Coccidioides Posadasii Research Articles

Disseminated Coccidioidomycosis Complicated by Bronchopulmonary Fistula and Viridans Streptococci Empyema

SESSION TITLE: Fungal Infections 1 SESSION TYPE: Affiliate Case Report Poster PRESENTED ON: Tuesday, October 31, 2017 at 01:30 PM - 02:30 PM INTRODUCTION: Coccidioidomycosis is typically a self-limited mild disease. Approximately 5-10% of patients with primary pulmonary coccidioidomycosis develop residual pulmonary sequelae such as fibrocavitary disease, which can rarely lead to bronchopulmonary fistulae (BPF). Although viridans streptococci (VS) has been documented to cause empyema, its coinfection in the setting of BPF complicating disseminated coccidioidomycosis infection has not been well described. CASE PRESENTATION: A 62-year-old Latino male farm worker with uncontrolled type II diabetes presented with acute left knee pain, progressively worsening over the course of three weeks. His history was also notable for a left arm nodule and a productive cough. Serum coccidioidal IgM was negative, IgG was positive, and complement fixation was 1:128. Synovial joint fluid from the left knee grew Coccidioides immitis and Coccidioides posadasii. Synovial tissue from the knee showed necrosis and multiple granulomata with coccidioidomycosis. Chest CT demonstrated a left-sided hydropneumothorax with associated BPF and pleural thickening. Additionally seen were innumerable bilateral centrilobular micronodules and left pleural-based cavity. Chest tube thoracostomy revealed purulent material and cultures with positive for VS. He subsequently underwent thoracoscopic debridement and decortication. Histopathology confirmed the presence of coccidioidomycosis. DISCUSSION: VS has been described the main virulent agent in community acquired pneumonia as well as in empyema and lung abscesses. Aspiration of oral secretions is hypothesized to be the etiology of VS respiratory infections. In our patient, rupture of the left-sided coccidial cavity abutting the pleura led to the development of a hydropneumothorax, and ultimately to the formation of a BPF, allowing for a contiguous pathway from the oral pharyngeal space into the pleural space. We hypothesize that the mechanism of VS empyema in our patient was via direct translocation of VS bacteria from the oral cavity through the fistula leading to inoculation of the coccidial pleural effusion. CONCLUSIONS: BPF is a rare complication of disseminated coccidioidomycosis infection. VS empyema has not been previously described in the setting of coccidioidomycosis infection and may be a more important pathogen in pleural pulmonary disease than previously recognized. A BPF may be a potential route of translocation for VS rather than aspiration when it is shown to be the sole pathogen in empyema. Reference #1: Thompson et al. Pulmonary Coccidioidomycosis. Semin Respir Crit Care Med 2011; 32(6): 754-763. 2011 Dec13 Reference #2: Choi SH, Cha SI, Choi KJ, Lim JK, Seo H, Yoo SS, Lee J, Lee SY, Kim CH, Park JY. Clinical Characteristics of Community-Acquired Viridans Streptococcal Pneumonia. Tuberc Respir Dis (Seoul). 2015 Jul;78(3):196-202. Reference #3: Jerng JS, Hsueh PR, Teng LJ, Lee LN, Yang PC, Luh KT. Empyema thoracis and lung abscess caused by viridans streptococci. Am J Respir Crit Care Med. 1997 Nov;156(5):1508-14. DISCLOSURE: The following authors have nothing to disclose: Margaret Wei, Nader Kamangar No Product/Research Disclosure Information

The Changing Epidemiology and Diagnosis of Valley Fever

Coccidioidomycosis, commonly known as valley fever, is a disease caused by two species of fungi, Coccidioides immitis and Coccidioides posadasii. Coccidioidomycosis is often self-limiting; however, in some patients, the disease can rapidly progress to a severe and potentially life-threatening illness. Proper diagnostics for coccidioidomycosis are important because acute disease can manifest as community-acquired pneumonia, and can be misdiagnosed as a viral or bacterial infection. Improper diagnosis can lead to unnecessary antibacterial therapy and may encourage extra-pulmonary proliferation of the fungus, which then requires longer antifungal therapy. Although coccidioidomycosis is caused by two different species, in terms of symptoms, disease progression, or clinical diagnostics, there are no known differences between the species. Additionally, recent work has revealed that the distribution of the organism in the environment may be changing and that the organism exists in the environment beyond the region of high endemicity. Overall, disease incidence has risen, and it is not known if this represents a better reporting and diagnosis infrastructure, increased environmental load, or increasing pathogenicity. Discussed here are the most common approaches to diagnose coccidioidomycosis, as well as recent advances in our understanding of the changing epidemiology of the disease and the causative organism.

Linear Epitopes of Paracoccidioides brasiliensis and Other Fungal Agents of Human Systemic Mycoses As Vaccine Candidates

Dimorphic fungi are agents of systemic mycoses associated with significant morbidity and frequent lethality in the Americas. Among the pathogenic species are Paracoccidioides brasiliensis and Paracoccidioides lutzii, which predominate in South America; Histoplasma capsulatum, Coccidioides posadasii, and Coccidioides immitis, and the Sporothrix spp. complex are other important pathogens. Associated with dimorphic fungi other important infections are caused by yeast such as Candida spp. and Cryptococcus spp. or mold such as Aspergillus spp., which are also fungal agents of deadly infections. Nowadays, the actual tendency of therapy is the development of a pan-fungal vaccine. This is, however, not easy because of the complexity of eukaryotic cells and the particularities of different species and isolates. Albeit there are several experimental vaccines being studied, we will focus mainly on peptide vaccines or epitopes of T-cell receptors inducing protective fungal responses. These peptides can be carried by antibody inducing β-(1,3)-glucan oligo or polysaccharides, or be mixed with them for administration. The present review discusses the efficacy of linear peptide epitopes in the context of antifungal immunization and vaccine proposition.

Update on the Epidemiology of Coccidioidomycosis

Coccidioidomycosis is an illness caused by the soil-dwelling, dimorphic fungi, Coccidioides immitis and Coccidioides posadasii, which are found primarily in niche ecological zones of the Western Hemisphere. The bulk of infections due to Coccidioides are found within the endemic areas of Arizona, California, Mexico, and Central America. Outcomes run the gamut from asymptomatic to a self-limited or even chronic pulmonary process, up to severe disseminated, and life-threatening disease. Patients at particular risk include the elderly, pregnant women, and members of certain ethnicities. Recent changes in the epidemiology and our overall understanding of coccidioidomycosis that pose a particular challenge to healthcare professionals include the rising incidence of disease, identification of infections thought to be acquired outside the previously described zones of endemicity, and the risks posed to the immunosuppressed population due to the increasing use of immunomodulatory pharmaceutical agents.

Preclinical identification of vaccine induced protective correlates in human leukocyte antigen expressing transgenic mice infected with Coccidioides posadasii

There is an emerging interest to develop human vaccines against medically-important fungal pathogens and a need for a preclinical animal model to assess vaccine efficacies and protective correlates. HLA-DR4 (DRB1∗0401 allele) transgenic mice express a human major histocompatibility complex class II (MHC II) receptor in such a way that CD4+ T-cell response is solely restricted by this human molecule. In this study HLA-DR4 transgenic mice were immunized with a live-attenuated vaccine (ΔT) and challenged by the intranasal route with 50–70 Coccidioides posadasii spores, a potentially lethal dose. The same vaccination regimen offers 100% survival for C57BL/6 mice. Conversely, ΔT-vaccinated HLA-DR4 mice displayed 3 distinct manifestations of Coccidioides infection including 40% fatal acute (FAD), 30% disseminated (DD) and 30% pulmonary disease (PD). The latter 2 groups of mice had reduced loss of body weight and survived to at least 50days postchallenge (dpc). These results suggest that ΔT vaccinated HLA-DR4 mice activated heterogeneous immunity against pulmonary Coccidioides infection. Vaccinated HLA-DR4 mice displayed early expansion of Th1 and Th17 cells and recruitment of inflammatory innate cells into Coccidioides-infected lungs during the first 9dpc. While contraction rates of Th cells and the inflammatory response during 14–35dpc significantly differed among the 3 groups of vaccinated HLA-DR4 mice. The FAD group displayed a sharply reduced Th1 and Th17 response, while overwhelmingly recruiting neutrophils into lungs during 9–14days. The FAD group approached moribund by 14dpc. In contrast, vaccinated HLA-DR4 survivors gradually contracted Th cells and inflammatory response with the greatest rate in the PD group. While vaccinated HLA-DR4 mice are susceptible to Coccidioides infection, they are useful for evaluation of vaccine efficacy and identification of immunological correlates against this mycosis.

A Coccidioides posadasii CPS1 Deletion Mutant Is Avirulent and Protects Mice from Lethal Infection.

The CPS1 gene was identified as a virulence factor in the maize pathogen Cochliobolus heterostrophus Hypothesizing that the homologous gene in Coccidioides posadasii could be important for virulence, we created a Δcps1 deletion mutant which was unable to cause disease in three strains of mice (C57BL/6, BALB/c, or the severely immunodeficient NOD-scid,γc(null) [NSG]). Only a single colony was recovered from 1 of 60 C57BL/6 mice following intranasal infections of up to 4,400 spores. Following administration of very high doses (10,000 to 2.5 × 10(7) spores) to NSG and BALB/c mice, spherules were observed in lung sections at time points from day 3 to day 10 postinfection, but nearly all appeared degraded with infrequent endosporulation. Although the role of CPS1 in virulence is not understood, phenotypic alterations and transcription differences of at least 33 genes in the Δcps1 strain versus C. posadasii is consistent with both metabolic and regulatory functions for the gene. The in vitro phenotype of the Δcps1 strain showed slower growth of mycelia with delayed and lower spore production than C. posadasii, and in vitro spherules were smaller. Vaccination of C57BL/6 or BALB/c mice with live Δcps1 spores either intranasally, intraperitoneally, or subcutaneously resulted in over 95% survival with mean residual lung fungal burdens of <1,000 CFU from an otherwise lethal C. posadasii intranasal infection. Considering its apparently complete attenuation of virulence and the high degree of resistance to C. posadasii infection when used as a vaccine, the Δcps1 strain is a promising vaccine candidate for preventing coccidioidomycosis in humans or other animals.

Synthesis and in vitro antifungal activity of isoniazid-derived hydrazones against Coccidioides posadasii

Coccidioidomycosis is a potentially severe infection caused by dimorphic fungi Coccidioides immitis and Coccidioides posadasii. Although guidelines are well established, refractory disease is a matter of concern in the clinical management of coccidioidomycosis. In the present study three isoniazid-derived hydrazones N′-[(E)-1-(4-methoxyphenyl)ethylidene]pyridine-4-carbohydrazide, N′-[(E)-1-(4-methylphenyl)ethylidene]pyridine-4-carbohydrazide, and N′-[(E)-1-(phenyl)ethylidene]pyridine-4-carbohydrazide were synthesized and evaluated for antifungal activity against C. posadasii. Susceptibility assays were performed by macrodilution testing. Interactions between the hydrazones and amphotericin B or itraconazole were evaluated by the checkerboard method. We also investigated the impairment of such compounds on cell ergosterol and membrane integrity. The synthesized molecules were able to inhibit C. posadasii in vitro with MIC values that ranged from 25 to 400 μg/mL. Drug interactions between synthesized molecules and amphotericin B proved synergistic for the majority of tested isolates; regarding itraconazole, synergism was observed only when strains were tested against N′-[(E)-1-(phenyl)ethylidene]pyridine-4-carbohydrazide. Reduction of cellular ergosterol was observed when strains were challenged with the hydrazones alone or combined with antifungals. Only N′-[(E)-1-(4-methylphenyl)ethylidene]pyridine-4-carbohydrazide altered membrane permeability of C. posadasii cells. Isoniazid-derived hydrazones were able to inhibit C. posadasii cells causing reduction of ergosterol content and alterations in the permeability of cell membrane. This study confirms the antifungal potential of hydrazones against pathogenic fungi.

Intact Immunotaxis Comprises an Intricate Spatiotemporal Hierarchy of Distinct Chemotactic Processes - A New Paradigm

A close look at cytokine-directed chemotaxis reveals an intriguing mechanistic dilemma. If cytokine gradients provided the only directional cue for chemotaxing immune cells, these cells would ultimately target the cytokine-producing cells of their own host. Such friendly fire may well be an inherent part of an indiscriminate mop-up duty of immune cells in the inflammatory response. However, it would preclude these cells from partaking in any pathogen-specific defenses requiring pre-contact recognition of invaders. Evidently, effective immune-cell recruitment requires additional directional guidance via chemical signals that emanate from the surfaces of microbes rather than host cells. To detect minuscule amounts of chemoattractant produced at the surfaces of bacteria, fungi, and surrogate particles, we have developed a single-cell, pure-chemotaxis assay that employs human neutrophils as ultrasensitive biodetectors. We use micropipette manipulation and/or optical tweezers to maneuver target particles into the proximity of non-adherent, initially quiescent neutrophils. Microscopic inspection of the resulting neutrophil morphology provides a clear readout of the chemotactic activity of the cells. In all cases where neutrophils extended chemotactic protrusions toward nearby targets, this cell response required serum. For bacterial targets such as Salmonella Typhimurium and Escherichia coli, and for zymosan particles as well as fungal particles of Coccidioides posadasii and Candida albicans, this type of microbe-guided chemotaxis was predominantly mediated by complement, in particular the anaphylatoxin C5a. The pre-contact neutrophil response to these targets was generally vigorous but short range, consistent with our analytical solution of a theoretical diffusion-reaction problem that takes into account the deactivation of anaphylatoxins by carboxypeptidases. Together, these findings establish complement-mediated chemotaxis as a universal homing mechanism by which chemotaxing immune cells implement a last-minute course correction toward bacterial, fungal, and model pathogens.

Evaluating Common Humoral Responses against Fungal Infections with Yeast Protein Microarrays.

We profiled the global immunoglobulin response against fungal infection by using yeast protein microarrays. Groups of CD-1 mice were infected systemically with human fungal pathogens (Coccidioides posadasii, Candida albicans, or Paracoccidioides brasiliensis) or inoculated with PBS as a control. Another group was inoculated with heat-killed yeast (HKY) of Saccharomyces cerevisiae. After 30 days, serum from mice in the groups were collected and used to probe S. cerevisiae protein microarrays containing 4800 full-length glutathione S-transferase (GST)-fusion proteins. Antimouse IgG conjugated with Alexafluor 555 and anti-GST antibody conjugated with Alexafluor 647 were used to detect antibody-antigen interactions and the presence of GST-fusion proteins, respectively. Serum after infection with C. albicans reacted with 121 proteins: C. posadasii, 81; P. brasiliensis, 67; and after HKY, 63 proteins on the yeast protein microarray, respectively. We identified a set of 16 antigenic proteins that were shared across the three fungal pathogens. These include retrotransposon capsid proteins, heat shock proteins, and mitochondrial proteins. Five of these proteins were identified in our previous study of fungal cell wall by mass spectrometry (Ann. N. Y. Acad. Sci. 2012, 1273, 44-51). The results obtained give a comprehensive view of the immunological responses to fungal infections at the proteomic level. They also offer insight into immunoreactive protein commonality among several fungal pathogens and provide a basis for a panfungal vaccine.

Whole glucan particles as a vaccine against systemic coccidioidomycosis.

We reported previously that yeast-derived whole glucan particles (WGPs), with or without conjugation to BSA, used as a vaccine protected against systemic aspergillosis in mice. Here, we examined their utility as a potential vaccine against coccidioidomycosis. WGPs were prepared from Saccharomyces cerevisiae; conjugation with BSA (WGP-BSA) was done using 1-cyano-4-dimethylaminopyridinium tetrafluoroborate-mediated conjugation. Heat-killed S. cerevisiae (HKY) was used as a positive-control vaccine. CD-1 mice were vaccinated with WGPs or WGP-BSA, HKY or PBS once weekly, beginning 21 days prior to infection. Mice were infected intravenously with arthroconidia of Coccidioides posadasii. In the low-mortality study, 50 % of PBS-treated controls died. Only WGP-BSA at 0.6 mg per dose induced significant protection compared with PBS treatment. All surviving mice were infected in all three organs examined. Those given WGP-BSA at 0.6 mg per dose had fewer c.f.u. in liver and lungs (P = 0.04), and those given WGPs at 6 mg per dose had fewer in lungs (P < 0.02), compared with PBS. In the high-mortality study, 90 % of PBS mice died. Vaccination with HKY, and WGPs or WGP-BSA at 6 or 12 mg per dose significantly prolonged survival (P ≤ 0.05). No surviving mice were free of infection. HKY and WGP-BSA at 12 mg per dose reduced c.f.u. in the liver and lungs (P < 0.05) and WGP-BSA at 6 mg per dose reduced c.f.u. in the lungs (P < 0.05); unconjugated WGPs did not reduce infection. WGPs or WGP-BSA acted as a vaccine that protected against mortality caused by coccidioidomycosis. Thus, WGP protection against coccidioidomycosis and aspergillosis provides the basis for development of a pan-fungal vaccine.

Coccidioides Endospores and Spherules Draw Strong Chemotactic, Adhesive, and Phagocytic Responses by Individual Human Neutrophils.

Coccidioides spp. are dimorphic pathogenic fungi whose parasitic forms cause coccidioidomycosis (Valley fever) in mammalian hosts. We use an innovative interdisciplinary approach to analyze one-on-one encounters between human neutrophils and two forms of Coccidioides posadasii. To examine the mechanisms by which the innate immune system coordinates different stages of the host response to fungal pathogens, we dissect the immune-cell response into chemotaxis, adhesion, and phagocytosis. Our single-cell technique reveals a surprisingly strong response by initially quiescent neutrophils to close encounters with C. posadasii, both from a distance (by complement-mediated chemotaxis) as well as upon contact (by serum-dependent adhesion and phagocytosis). This response closely resembles neutrophil interactions with Candida albicans and zymosan particles, and is significantly stronger than the neutrophil responses to Cryptococcus neoformans, Aspergillus fumigatus, and Rhizopus oryzae under identical conditions. The vigorous in vitro neutrophil response suggests that C. posadasii evades in vivo recognition by neutrophils through suppression of long-range mobilization and recruitment of the immune cells. This observation elucidates an important paradigm of the recognition of microbes, i.e., that intact immunotaxis comprises an intricate spatiotemporal hierarchy of distinct chemotactic processes. Moreover, in contrast to earlier reports, human neutrophils exhibit vigorous chemotaxis toward, and frustrated phagocytosis of, the large spherules of C. posadasii under physiological-like conditions. Finally, neutrophils from healthy donors and patients with chronic coccidioidomycosis display subtle differences in their responses to antibody-coated beads, even though the patient cells appear to interact normally with C. posadasii endospores.

Coccidioides species determination: does sequence analysis agree with restriction fragment length polymorphism?

Fifteen Coccidioides isolates were previously examined for genetic diversity using restriction fragment length polymorphism (RFLP); two fragment patterns were observed. Two isolates demonstrated one banding pattern (designated RFLP group I), while the remaining 13 isolates demonstrated a second pattern (designated RFLP group II). Recently, molecular studies supported the division of the genera Coccidioides into two species: Coccidioides posadasii and Coccidioides immitis. It has been assumed that the species division corresponds to the RFLP grouping. We tested this hypothesis by amplifying the ribosomal DNA internal transcribed spacer region as well as the dioxygenase, serine proteinase, and urease genes from 13 isolates previously examined by RFLP and then sequencing the PCR products. The appropriate species for each isolate was assigned using phylogenetically informative sites. The RFLP grouping agreed with the Coccidioides species assignment for all but one isolate, which may represent a hybrid. In addition, polymorphic sites among the four genes examined were in agreement for species assignment such that analysis of a single gene may be sufficient for species assignment.

Identification, molecular characterization, and expression analysis of a DOMON-like type 9 carbohydrate-binding module domain-containing protein of Coccidioides posadasii.

Previously, we investigated the effect of N-acetylglucosamine (GlcNAc) on Coccidioides posadasii chitinolytic enzymes during in vitro spherule-endospore (S/E) phase culture. During those studies, sodium dodecyl sulfatepolyacrylamide gel electrophoresis analysis of supernatants from S/E phase cultures grown in Converse medium with or without added GlcNAc revealed a ∼ 28-kDa band (CFP28), whose abundance was increased by GlcNAc in parallel with the chitinolytic enzymes. Mass spectrometry (MS) of the CFP28 band revealed peptides that matched an open reading frame found in the tentative consensus sequence, TC20325, retrieved from the Dana Farber Cancer Institute C. posadasii Gene Index Database. The TC20325 cDNA sequence was used to design internal primers based on MS peptides and a full-length cDNA was isolated using a combination of rapid amplification of cDNA ends and reverse transcription-polymerase chain reaction. The deduced amino acid sequence of the full-length cDNA consists of 231 amino acid residues with a 19 aa signal peptide. The mature protein has a calculated molecular mass of ∼ 24.5 kDa, a theoretical pI of 6.09, and consists of a single DOMON-like type 9 carbohydrate-binding module (CBM9-like-3) conserved domain. The protein shares the highest sequence similarity (≥57%) to hypothetical proteins from fungi within the Pezizomycotina subphylum of Ascomycota. Antiserum against a recombinant version of CFP28 recognized native CFP28 in S/E phase cells and culture supernatants. CFP28 mRNA and protein expression were detectable in S/E phase in Converse medium, but were increased in the presence of added GlcNAc. Purified native CFP28 reacted with pooled sera from patients with coccidioidomycosis.

Laboratory methods in the diagnosis of coccidioidomycosis

Coccidioides immitis and Coccidioides posadasii, the two recognized causes of coccidioidomicosis, may be diagnosed by direct microscopy, culture, serologic and the skin-test with mycelial coccidioidin. Identification of spherules by direct examination of secretions after addition of KOH, is more rapid and may speed diagnosis. Histopathology of infected tissue demonstrates acute and chronic granulomatous inflammation and in the section strained with hematoxylin-eosine (HE) or Grocott metamine-silver (GMS). Isolation of Coccidioides spp. by culture is not difficult, the fungus grows well on most mycologic media after 5-7 days of incubation and the laboratory should maintain a biological safety level 2 or 3. Phenotypic identification to genus level may be achieved by visualization of arthroconidia in culture. Isolates may be confirmed as Coccidioides ssp. by molecular probes, but separation of species is best achieved by specialized molecular techniques, available at the Reference Laboratory. Immune diffusion and enzyme-immune assays (EIA) are commonly used for detection of IgM and IgG antibodies in serum. Sequential complement fixations (CF) for IgG class of antibody are useful for the prognosis of coccidioidomycosis.

Development and validation of a quantitative real-time PCR assay for the early diagnosis of coccidioidomycosis

A new real-time polymerase chain reaction (RT-PCR) assay based on a Coccidioides genus–specific molecular beacon probe was developed for the detection of coccidioidomycosis and validated with tissues from animal models and clinical samples. The assay showed high analytic reproducibility (r2 > 0.99) and specificity for cultured strains (100%); the lower limit of detection was 1 fg of genomic DNA/μL of reaction. Fungal burdens in the organs of mice infected with Coccidioides posadasii strain Silveira were more accurately quantified by RT-PCR compared to colony-forming unit for all tissues. The RT-PCR assay was positive for 97.7% of spleen and 100% of liver or lung. Progression of infection in all organs was similar by both methods (P > 0.05). The sensitivity of the assay also was 100% for paraffin-embedded samples and samples from patients with positive cultures. Our RT-PCR assay is effective for the diagnosis and monitoring of Coccidioides infection, and its use also avoids the biohazard and time delay of identifying cultures in the clinical setting.

INTESTINAL COCCIDIOIDOMYCOSIS IN A RED COACHWHIP SNAKE (MASTICOPHIS FLAGELLUM PICEUS)

An adult female, wild-caught red coachwhip snake (Masticophis flagellum piceus) was euthanized at the Phoenix Zoo due to severe neurologic signs. Necropsy and histopathology revealed an invasive liposarcoma of the vertebral column, which likely caused the neurologic signs. Histology of the small intestine revealed a granuloma with intralesional yeasts morphologically compatible with the genus Coccidioides. The diagnosis of coccidioidomycosis was confirmed with immunohistochemistry staining. Coccidioides posadasii is endemic to Arizona and is an important cause of disseminated fungal infections in mammals in this region. This is the first known report of intestinal coccidioidomycosis in a veterinary species and the second report of coccidioidomycosis in a reptile.

Coccidioidomycosis masquerading as skeletal tuberculosis: an imported case and review of coccidioidomycosis in India

We describe a possible imported case of osteo-articular coccidioidomycosis in India. Culture of the computed tomography-guided aspirate revealed the growth of Coccidioides spp., which was identified as Coccidioides posadasii by sequencing of the internal transcribed spacer (ITS) region of rDNA. He was successfully treated with amphotericin B followed by itraconazole. All the previous published reports of coccidioidomycosis cases diagnosed in India are also reviewed in order to increase the awareness of this disease in non-endemic areas.

Molecular markers in the epidemiology and diagnosis of coccidioidomycosis

The prevalence of coccidioidomycosis in endemic areas has been observed to increase daily. To understand the causes of the spread of the disease and design strategies for fungal detection in clinical and environmental samples, scientists have resorted to molecular tools that allow fungal detection in a natural environment, reliable identification in clinical cases and the study of biological characteristics, such as reproductive and genetic structure, demographic history and diversification. We conducted a review of the most important molecular markers in the epidemiology of Coccidioides spp. and the diagnosis of coccidioidomycosis. A literature search was performed for scientific publications concerning the application of molecular tools for the epidemiology and diagnosis of coccidioidomycosis. The use of molecular markers in the epidemiological study and diagnosis of coccidioidomycosis has allowed for the typing of Coccidioides spp. isolates, improved understanding of their mode of reproduction, genetic variation and speciation and resulted in the development specific, rapid and sensitive strategies for detecting the fungus in environmental and clinical samples. Molecular markers have revealed genetic variability in Coccidioides spp. This finding influences changes in the epidemiology of coccidioidomycosis, such as the emergence of more virulent or antifungal resistant genotypes. Furthermore, the molecular markers currently used to identify Coccidioides immitis and Coccidioides posadasii are specific and sensitive. However, they must be validated to determine their application in diagnosis.This manuscript is part of the series of works presented at the “V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi” (Oaxaca, Mexico, 2012).

Kokzidioidomykose mit Befall der Zunge

Systemic mycoses are rare but important differential diagnoses in patients with imported infections. We report the case of a 51-year-old German traveller who acquired coccidioidomycosis during a holiday in Arizona, USA. The disease became apparent several months later primarily as a swelling of the tongue. Subsequent diagnostic investigations revealed infiltration of both lungs. The causal agent Coccidioides posadasii could be cultivated from transbronchial biopsy samples.

Anti-inflammatory and Immunomodulatory Effect of an Extract of Coccidioides posadasii in Experimental Arthritis

Trying to surpass host defenses, fungal infections alter the immune response. Components from nonpathogenic fungi present therapeutic anti-inflammatory and immunomodulating activities. This study reveals that proteins present in a Coccidioides posadasii extract provide anti-inflammatory benefit in experimental arthritis. Zymosan was given intra-articularly to rats and mice, and groups were pretreated with C. posadasii extract either per os or intraperitoneally. Controls received the vehicle. Acute hypernociception was evaluated using articular incapacitation and von Frey methods. Cell influx and cytokine levels were assessed in joint exudates. Joint damage was evaluated by histopathology and determination of glycosaminoglycan content of the cartilage. Synovia was evaluated for cell death and inducible nitric oxide synthase (iNOS) expression using TUNEL and immunohistochemistry, respectively. Pretreatment with C. posadasii extract significantly inhibited acute and chronic cell influx, hypernociception, and provoked reduction of glycosaminoglycan loss while reducing chronic synovitis, cell death, and iNOS expression. Reduction/alkylation of C. posadasii extract abrogated these effects. C. posadasii administration did not alter TNF-α, IL-1β, IL-17, and γ-interferon levels, whereas IL-10 levels were significantly reduced. Data reveal that a C. posadasii extract reduces iNOS expression that is associated with inhibition of synovial apoptosis and decrease in IL-10 levels released into zymosan-inflamed joints. Characterization of active components excluded charged carbohydrates while pointing to a protein as responsible for these effects. In summary, systemic administration of components from a pathogenic fungus provides anti-inflammatory effects, being species-independent and orally active. Besides adding to understand host response against fungi, the results may lead to therapeutic implications.