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Atomic force microscopy of Cryptococcus secreted polysaccharides (secreted-PS). Topographical images obtained at PeakForce tapping mode (in air) of C. neoformans (A and C) and C. liquefaciens (B and D) secreted-PS. Arrowheads indicate handle-shaped structures branching off polysaccharide fibers. Scale bars: 300 nm (A,B) and 100 nm (C,D).
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Invasive fungal infections, including cryptococcosis, are a growing threat to immunocompromised patients. Although Cryptococcus neoformans and Cryptococcus gattii are the main agents of human cryptococcosis, opportunistic infections by environmental species, such as C. liquefaciens, have been observed recently. The main Cryptococcus virulence facto...
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Context 1
... examined the organization of secreted-PS molecules from both species using atomic force microscopy (AFM), which allowed us to obtain topographic images in the nanometer scale without critical point drying. AFM images of secreted-PS showed linear molecules with average length of 675 ± 14 nm, for C. neoformans, and 704 ± 44 nm, for C. liquefaciens (Fig. 2). Branch 'handles' -globular structures at one end of each fiber (arrow- heads in Fig. 2), similar to those previously described in branched commercial purified PS visualized by AFM 36 - were more common in C. neoformans PS, but were also present in C. liquefaciens PS (Fig. ...
Context 2
... (AFM), which allowed us to obtain topographic images in the nanometer scale without critical point drying. AFM images of secreted-PS showed linear molecules with average length of 675 ± 14 nm, for C. neoformans, and 704 ± 44 nm, for C. liquefaciens (Fig. 2). Branch 'handles' -globular structures at one end of each fiber (arrow- heads in Fig. 2), similar to those previously described in branched commercial purified PS visualized by AFM 36 - were more common in C. neoformans PS, but were also present in C. liquefaciens PS (Fig. ...
Context 3
... of 675 ± 14 nm, for C. neoformans, and 704 ± 44 nm, for C. liquefaciens (Fig. 2). Branch 'handles' -globular structures at one end of each fiber (arrow- heads in Fig. 2), similar to those previously described in branched commercial purified PS visualized by AFM 36 - were more common in C. neoformans PS, but were also present in C. liquefaciens PS (Fig. ...
Context 4
... we demonstrated that there are significant physico-chemical differences between secreted and capsular PS in C. neoformans 42 , showing that these molecules have distinct properties and should, therefore, be analyzed independently. AFM images of secreted PS from C. liquefaciens showed linear fibers containing a handle-like structure at the end (Fig. 2), similar to that described for C. neoformans 28,43 . For both species, although PS fibers were linear, as expected for these sugars, fibers interacted to form the aggregated, branched structures that were observed in images of both secreted and capsular ...
Similar publications
The genus Cryptococcus is composed of encapsulated yeasts that have the ability to infect and cause disease in humans, as Cryptococcus gattii/Cryptococcus neoformans species complex. Facing the current research panel, few studies about cryptococcosis and epidemiological data have been carried out in the western region of Sao Paulo state. This study...
Cryptococcosis, a systemic mycosis capable of disseminating to the central nervous system with frequent lethal effects, is caused by the species Cryptococus neoformans and Cryptococcus gattii. Several infectious agents such as virus, bacteria, and parasites may be associated to DNA damage and carcinogenesis in humans. Products of the oxidative meta...
Cryptococcosis, one of the most important systemic mycosis in the world, is caused by different genotypes of Cryptococcus neoformans and Cryptococcus gattii, which differ in their ecology, epidemiology, and antifungal susceptibility. Therefore, the search for new molecular markers for genotyping, pathogenicity and drug susceptibility is necessary....
Cryptococcus neoformans and Cryptococcus gattii are the etiologic agents of cryptococcosis, whose suitable treatment depends of rapid and correct detection and differentiation of the Cryptococcus specie. Currently, this identification is made by classical and molecular techniques, however most of them are considered laborious and expensive. As an a...
Introduction:
This study aimed to describe cryptococcal meningitis (CM) cases and the associated demographic, clinical, and microbiological data obtained from cities in the State of Mato Grosso do Sul in the Midwestern region of Brazil.
Methods:
The data from 129 patients with laboratory-confirmed CM admitted from 1997 to 2014 were retrospective...
Citations
... The emergence of pathogenic yeasts continues to threaten humanity, particularly individuals with underlying health conditions [1]. Among these yeast species, the Cryptococcus neoformans and Cryptococcus gattii species (CnCg species) complexes contain the most important and best-characterized pathogens [2]. ...
Emerging environmental pathogenic fungal infections, including cryptococcosis, continue to pose a significant threat to humans with compromised immunity and, to some extent, healthy ones. Cryptococcus neoformans was originally identified as the main etiological agent of human cryptococcosis, but recent studies have also identified the occurrence of opportunistic infections caused by Cryptococcus gattii . These two saprophytic facultative yeasts present a paradox as they can infect humans without requiring a host for replication or survival, a phenomenon termed readymade virulence. Many cryptococcal virulence traits appear to have dual effects that provide survival advantages in both animal hosts and the environment. Several molecular techniques have been developed to provide in-depth knowledge of these species complexes. This review will focus on the description of the Cryptococcus neoformans and Cryptococcus gattii ( CnCg ) species complexes and associated cryptococcal pathogenesis, ecological niches, and virulence factors employed by the pathogens to cause disease.
... After fixation, cells were washed in 0.1 M sodium cacodylate buffer (pH 7.2) containing 0.2 M sucrose and 2 mM MgCl 2 . Cell surface was observed by scanning electron microscopy (SEM) [35]. Obtained images were colored using Photoshop software. ...
The phenotypic plasticity of Cryptococcus neoformans is widely studied and demonstrated in vitro, but its influence on pathogenicity remains unclear. In this study, we investigated the dynamics of cryptococcal cell and transcriptional remodeling during pulmonary infection in a murine model. We showed that in Cryptococcus neoformans, cell size reduction (cell body ≤ 3 µm) is important for initial adaptation during infection. This change was associated with reproductive fitness and tissue invasion. Subsequently, the fungus develops mechanisms aimed at resistance to the host’s immune response, which is determinant for virulence. We investigated the transcriptional changes involved in this cellular remodeling and found an upregulation of transcripts related to ribosome biogenesis at the beginning (6 h) of infection and a later (10 days) upregulation of transcripts involved in the inositol pathway, energy production, and the proteasome. Consistent with a role for the proteasome, we found that its inhibition delayed cell remodeling during infection with the H99 strain. Altogether, these results further our understanding of the infection biology of C. neoformans and provide perspectives to support therapeutic and diagnostic targets for cryptococcosis.
... Several virulence factors have classically been carefully characterized and validated for C. gattii/neoformans complex, such as the presence of a polysaccharide capsule that protects yeast cells against phagocytosis, the ability to grow at 37°C, biofilm formation, and the production of additional factors including melanin, phospholipase, metalloprotease, DNase, urease, superoxide dismutase and other antioxidant enzymes (Mancianti et al., 2002;Sanchez et al., 2008;Peng et al., 2018;Zaragoza, 2019). However, only a few studies have investigated the potential role of these and other virulence factors in the non-Cryptococcus gattii/neoformans species complex (Ikeda et al., 2002;Andrade-Silva et al., 2010;Araujo Gde et al., 2012;Ferreira-Paim et al., 2012;Araujo et al., 2017). ...
... The non-Cryptococcus gattii/neoformans species complex strains (clades III to V) had markedly thinner capsules. However, Araujo et al., (Araujo et al., 2017) described that polysaccharides of C. liquefaciens have strikingly similar ultrastructural and biological properties to those of C. neoformans, and found that this species led to mortality rates similar to C. neoformans in a virulence model with the larvae of Galleria melonella, reinforcing its pathogenic potential. ...
Cryptococcosis is an infectious disease of worldwide distribution, caused by encapsulated yeasts belonging to the phylum Basidiomycota. The genus Cryptococcus includes several species distributed around the world. The C. gattii/neoformans species complex is largely responsible for most cases of cryptococcosis. However, clinical series have been published of infections caused by Papiliotrema (Cryptococcus) laurentii and Naganishia albida (Cryptococcus albidus), among other related genera. Here, we examined the pathogenic potential and antifungal susceptibility of C. gattii/neoformans species complex (clades I and II) and related genera (Papiliotrema and Naganishia) isolated from environmental and clinical samples. P. laurentii (clade III), N. liquefasciens/N. albidosimilis (clade IV); and N. adeliensis/N. albida (clade V) strains produced higher levels of phospholipase and hemolysins, whereas the C. gattii/neoformans species complex strains (clades I and II) had markedly thicker capsules, produced more biofilm biomass and melanin, which are known virulence attributes. Interestingly, 40% of C. neoformans strains (clade II) had MICs above the ECV established for this species to amphotericin B. Several non-C. gattii/neoformans species complex (clades III to V) had MICs equal to or above the ECVs established for C. deuterogattii and C. neoformans for all the three antifungal drugs tested. Finally, all the non-C. gattii/neoformans clinical isolates (clades III to V) produced more melanin than the environmental isolates might reflect their particularly enhanced need for melanin during in vivo protection. It is very clear that C. gattii/neoformans species complex (clades I and II) strains, in general, show more similar virulence phenotypes between each other when compared to non-C. gattii/neoformans species complex (clades III to V) isolates. These observations together with the fact that P. laurentii and Naganishia spp. (clades III to V) strains were collected from the outside of a University Hospital, identify features of these yeasts important for environmental and patient colonization and furthermore, define mechanisms for infections with these uncommon pathogens.
... developed several adaptation mechanisms, known as virulence factors. Some examples are (i) melanin production and cell wall remodeling (resistance to cell-mediated death and immunomodulation) (Huffnagle et al., 1995;Wang et al., 1995;Doering et al., 1999;Liu et al., 1999;Gómez and Nosanchuk, 2003), (ii) production of superoxide dismutase (protection against toxic free radicals) (Cox et al., 2003), (iii) phospholipase and urease secretion (intracellular growth, diffusion and proliferation) (Cox et al., 2000(Cox et al., , 2001, (iv) phenotypic switching (immune evasion) (Goldman et al., 1998;Fries et al., 2001), (v) cellular gigantism (immune evasion) (Okagaki et al., 2010;Zaragoza and Nielsen, 2013;Trevijano-Contador et al., 2018;Zaragoza, 2019), (vi) thermotolerance (Johnston et al., 2016;Araújo et al., 2017;Bloom et al., 2019), and (vii) PS production, which is the main virulence factor used by C. neoformans (Zaragoza et al., 2009;Araujo et al., 2012;Zaragoza, 2019). Most of these features are believed to have been acquired through selective pressures and are likely to be the result of interactions with environmental predators, such as amoebae and nematodes (Casadevall and Pirofski, 2007;Albuquerque et al., 2019). ...
... It is known, for example, that the capsule matrix exhibits clear vertical stratification, with distinct density regions, with its inner part having a higher fiber density Frontiers in Microbiology | www.frontiersin.org than its outer region (Gates et al., 2004;Bryan et al., 2005;Frases et al., 2009b;Araújo et al., 2016Araújo et al., , 2017. Although softer than the cell wall and presenting viscoelastic properties (Frases et al., 2009a;Araújo et al., 2019), the high PS density of the inner region prevents the penetration of larger macromolecules, including antibodies and proteins of the complement system, restricting the access of these molecules to the cell wall (Gates et al., 2004;Gates and Kozel, 2006). ...
Cryptococcus neoformans is a fungal pathogen that causes life-threatening infections in immunocompromised individuals. It is surrounded by three concentric structures that separate the cell from the extracellular space: the plasma membrane, the cell wall and the polysaccharide (PS) capsule. Although several studies have revealed the chemical composition of these structures, little is known about their ultrastructural organization and remodeling during C. neoformans budding events. Here, by combining the latest and most accurate light and electron microscopy techniques, we describe the morphological remodeling that occurs among the capsule, cell wall and plasma membrane during budding in C. neoformans. Our results show that the cell wall deforms to generate a specialized region at one of the cell’s poles. This region subsequently begins to break into layers that are slightly separated from each other and with thick tips. We also observe a reorganization of the capsular PS around the specialized regions. While daughter cells present their PS fibers aligned in the direction of budding, mother cells show a similar pattern but in the opposite direction. Also, daughter cells form multilamellar membrane structures covering the continuous opening between both cells. Together, our findings provide compelling ultrastructural evidence for C. neoformans surface remodeling during budding, which may have important implications for future studies exploring these remodeled specialized regions as drug-targets against cryptococcosis.
... To assess possible influences on the virulence of C. neoformans after treatments with either DX or MP, we utilized the invertebrate host G. mellonella. This model has been used to study not only C. neoformans virulence but also the action of antifungals against this pathogen (Araujo et al., 2012;Araújo et al., 2017). All infected and untreated larvae died by day 8 upon infection (Figure 4). ...
... In contrast, the MP treatment did not induce a significant change in PS production but showed significant alterations in the electronegativity of secreted PS, with more negative Zeta potentials compared to control cells, and size of fragments, with values 2.6-times smaller in terms of effective diameter. Previous results from our group have demonstrated that the biological properties of PSs are greatly influenced by their size and physicochemical properties, implying that structural parameters can not only alter their biological functions but can also induce different host responses (Frases et al., 2009a,b;Albuquerque et al., 2014;Pontes and Frases, 2015;Araújo et al., 2016Araújo et al., , 2017Araújo et al., , 2019. Overall, our results suggest that changes in the size, morphology, and electronegativity of the capsule and secreted PSs can affect the fungal pathogenesis, producing different dissemination patterns due to the influence of corticosteroids in these structures. ...
... In addition to the effects that these drugs have on the host's phagocytic cells, we are showing in the present study that DX and MP also influence the secreted capsule and polysaccharide sizes and electronegativity. Changes in capsule size, flexibility and electronegativity have been described as capable of influencing the capacity of macrophages to phagocytize C. neoformans (Frases et al., 2011;Albuquerque et al., 2014;Pontes and Frases, 2015;Ding et al., 2016;Araújo et al., 2017). Thus, we conjecture that the effects of DX and MP would probably reduce the ability of macrophages to phagocytize C. neoformans, not only due to the direct effects of these drugs on macrophages (Balow and Rosenthal, 1973;Keil et al., 1995;Xie et al., 2019), but also because the alterations in the fungus surface would probably lead to a decrease in phagocytosis if compared to the changes already reported in the literature (Frases et al., 2011;Albuquerque et al., 2014;Pontes and Frases, 2015;Ding et al., 2016;Araújo et al., 2017). ...
Cryptococcus neoformans is a fungal pathogen that causes life-threatening infections in immunocompromised individuals, who often have some inflammatory condition and, therefore, end up using glucocorticoids, such as dexamethasone and methylprednisolone. Although the effects of this class of molecules during cryptococcosis have been investigated, their consequences for the biology of C. neoformans is less explored. Here, we studied the effects of dexamethasone and methylprednisolone on the metabolism and on the induction of virulence factors in C. neoformans . Our results showed that both glucocorticoids increased fungal cell proliferation and surface electronegativity but reduced capsule and secreted polysaccharide sizes, as well as capsule compaction, by decreasing the density of polysaccharide fibers. We also tested whether glucocorticoids could affect the fungal virulence in Galleria mellonella and mice. Although the survival rate of Galleria larvae increased, those from mice showed a tendency to decrease, with infected animals dying earlier after glucocorticoid treatments. The pathogenesis of spread of cryptococcosis and the interleukin secretion pattern were also assessed for lungs and brains of infected mice. While increases in the spread of the fungus to lungs were observed after treatment with glucocorticoids, a significant difference in brain was observed only for methylprednisolone, although a trend toward increasing was also observed for dexamethasone. Moreover, increases in both pulmonary and cerebral IL-10 production, reduction of IL-6 production but no changes in IL-4, IL-17, and INF-γ were also observed after glucocorticoid treatments. Finally, histopathological analysis confirmed the increase in number of fungal cells in lung and brain tissues of mice previously subjected to dexamethasone or methylprednisolone treatments. Together, our results provide compelling evidence for the effects of dexamethasone and methylprednisolone on the biology of C. neoformans and may have important implications for future clinical treatments, calling attention to the risks of using these glucocorticoids against cryptococcosis or in immunocompromised individuals.
... Besides Cryptococcus spp (Araujo et al., 2017), another Basidiomycota are also able to produce GXM-like molecules. Trichosporon asahii produces a functional GXM-like molecule with similar glycosyl composition to cryptococcal GXM that also manifests antiphagocytic activities (Fonseca et al., 2009). ...
... MAb 18B7 labeling ranged from a dotted to a ring pattern, confirming previous indications by our group (Cordero et al., 2016). This same binding pattern was originally reported for Cn (Casadevall et al., 1998) and other GXM-like polysaccharide producing fungi, such as C. liquefaciens (Araujo et al., 2017), T. asahii (Fonseca et al., 2009), T. mucoides (Zimbres et al., 2018) and P. brasiliensis (Albuquerque et al., 2012). ...
... As a GXM-like component, C-gly-Hc could be promptly incorporated by the Cn cap59 acapsular mutant, forming a capsule-like structure, with some dotted regions, resembling the capsule formed when C-gly-Cn was used. A similar profile was also observed for the incorporation of other GXM-like components of the most diverse origin, likely indicating a shared property among them (Fonseca et al., 2009;Albuquerque et al., 2012;Araujo et al., 2017;Zimbres et al., 2018). As for Cn GXM, C-gly-Hc incorporation could occur via a-1,3-glucan (Reese and Doering, 2003) or attachment to cell wall chitin . ...
The cell wall is a ubiquitous structure in the fungal kingdom, with some features varying depending on the species. Additional external structures can be present, such as the capsule of Cryptococcus neoformans (Cn), its major virulence factor, mainly composed of glucuronoxylomannan (GXM), with anti-phagocytic and anti-inflammatory properties. The literature shows that other cryptococcal species and even more evolutionarily distant species, such as the Trichosporon asahii, T. mucoides, and Paracoccidioides brasiliensis can produce GXM-like polysaccharides displaying serological reactivity to GXM-specific monoclonal antibodies (mAbs), and these complex polysaccharides have similar composition and anti-phagocytic properties to cryptococcal GXM. Previously, we demonstrated that the fungus Histoplasma capsulatum (Hc) incorporates, surface/secreted GXM of Cn and the surface accumulation of the polysaccharide enhances Hc virulence in vitro and in vivo. In this work, we characterized the ability of Hc to produce cellular-attached (C-gly-Hc) and secreted (E-gly) glycans with reactivity to GXM mAbs. These C-gly-Hc are readily incorporated on the surface of acapsular Cn cap59; however, in contrast to Cn GXM, C-gly-Hc had no xylose and glucuronic acid in its composition. Mapping of recognized Cn GXM synthesis/export proteins confirmed the presence of orthologs in the Hc database. Evaluation of C-gly and E-gly of Hc from strains of distinct monophyletic clades showed serological reactivity to GXM mAbs, despite slight differences in their molecular dimensions. These C-gly-Hc and E-gly-Hc also reacted with sera of cryptococcosis patients. In turn, sera from histoplasmosis patients recognized Cn glycans, suggesting immunogenicity and the presence of cross-reacting antibodies. Additionally, C-gly-Hc and E-gly-Hc coated Cn cap59 were more resistant to phagocytosis and macrophage killing. C-gly-Hc and E-gly-Hc coated Cn cap59 were also able to kill larvae of Galleria mellonella. These GXM-like Hc glycans, as well as those produced by other pathogenic fungi, may also be important during host-pathogen interactions, and factors associated with their regulation are potentially important targets for the management of histoplasmosis.
... and C. neoformans sensu lato have close similarities in their structural composition, especially in the mucopolysaccharide capsule. Thus, the importance of the study of this species is justified, since they have virulence factors similar to those already considered for pathogenic species [39,40]. ...
Introduction:
Fungi of the genus Cryptococcus are cosmopolitan and may be agents of opportunistic mycoses in immunocompromised and sometimes immunocompetent individuals. Cryptococcus species are frequently isolated from trees and bird excreta in the environment and infection occurs by inhalation of propagules dispersed in the air. The aim was to investigate Cryptococcus species in bird excreta and tree hollows located in a university hospital area and in an academic area of a university campus.
Methodology:
A total of 40 samples of bird excreta and 41 samples of tree hollows were collected. The identification of the isolates was done by classical methodology and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
Results:
Twenty (62.5%) isolates of Cryptococcus were found in bird excreta and 12 (37.5%) in tree hollows. C. laurentii (currently Papiliotrema laurentii) was the most frequent species in both samples, being found in 5 samples of excreta and in 8 tree hollows. The diversity of species found in excreta (C. laurentii, C. albidus [currently Naganishia albida], C. liquefaciens [currently N. liquefaciens], C. friedmanii [currently N. friedmannii] and others) was higher than in tree hollows (C. laurentii, C. flavescens [currently Papiliotrema flavescens], and other yeasts).
Conclusion:
Many Cryptococcus species were isolated from excreta and tree hollows, and this fact is important for understanding the environmental epidemiology of those emerging pathogens for public health, as a way to implement surveillance actions and control of cryptococcosis.
... Studies on cryptococcal virulence factors are typically conducted in C. neoformans, since other species of the genus were presumed to have low pathogenic potential. Recently, Araujo and co-workers (2017) described striking similarities in composition and structure between PS molecules of C. liquefaciens and C. neoformans, and showed that PS molecules from these species have comparable behavior in several key biological activities [72,73]. The remarkable similarities in capsule ultrastructure and virulence between single isolates of C. neoformans and C. liquefaciens described in the study of Araujo and co-workers (2017), together with the first descriptions of human fungemia by C. liquefaciens in immunocompromised patients [70][71][72][73][74], suggest that this environmental cryptococcal species may represent a novel human pathogen. ...
... Recently, Araujo and co-workers (2017) described striking similarities in composition and structure between PS molecules of C. liquefaciens and C. neoformans, and showed that PS molecules from these species have comparable behavior in several key biological activities [72,73]. The remarkable similarities in capsule ultrastructure and virulence between single isolates of C. neoformans and C. liquefaciens described in the study of Araujo and co-workers (2017), together with the first descriptions of human fungemia by C. liquefaciens in immunocompromised patients [70][71][72][73][74], suggest that this environmental cryptococcal species may represent a novel human pathogen. Importantly, C. liquefaciens infections are resistant to 5-fluorocytosine, indicating that that treatment of fungemia by this species may be challenging [70,71]. ...
Invasive fungal infections are a growing threat to immunocompromised patients, highlighting the importance of monitoring fungal pathogens. Global warming (including climatic oscillations) may select for environmental species that have acquired thermotolerance, a key step toward pathogenesis to humans. Also, important virulence factors have developed in environmental fungi, because they are essential for yeast survival in the environment. Thus, fungi traditionally regarded as nonpathogenic to humans have virulence factors similar to those of their pathogenic relatives. Here, we highlight the emergence of saprophytic environmental fungi - including species of Cryptococcus, Aspergillus, Penicillium, Candida and Scedosporium - as new human pathogens. Emerging pathogens are, in some cases, resistant to the available antifungals, potentiating the threat of novel fungal diseases.
This article reports an elderly male patient with nodules and ulcers on the face and behind the left ear after trauma. Primary cutaneous cryptococcosis was confirmed using pathological biopsy, special staining, tissue culture, and fungal sequencing. The patient received a therapeutic intervention involving the administration of the antifungal agent itraconazole. Substantial amelioration of cutaneous manifestations was observed after a 3-month course of treatment. After an elapsed interval, the patient was diagnosed with esophageal tumor. Moreover, the literature on 33 patients with primary cutaneous cryptococcosis published in the past 10 years was also reviewed.
The Cryptococcus genus comprises more than 100 species, of which C. neoformans and C. gattii are the leading cause of cryptococcosis. The distribution of C. gattii and C. neoformans species complexes has been extensively studied and widely reported globally. Other species such as Naganishia albida, Papiliotrema laurentii, and Papiliotrema flavescens have been reported as pathogenic yeasts. Since there are no reports of environmental isolation in the Boyacá region (Colombia), this study aimed to isolate and characterize Cryptococcus and Cryptococcus-like yeasts from pigeon feces, Eucalyptus, and olive trees distributed in the municipalities of Tunja and Ricaute Alto. The environmental data was recovered, and the isolations obtained were identified by microscopy, biochemical test, MALDI-TOF MS, URA5-RFLP, and sequencing of the ITS and LSU loci. For the 93 pigeon dropping samples collected in Tunja, 23 yielded to C. neoformans, 3 to N. globosa, 2 N. albida and 1 to P. laurentii. Of the 1188 samples collected from olive trees, 17 (1.43%) positive samples were identified as C. gattii species complex (4), C. neoformans species complex (2), P. laurentii (3), N. albida (2), N. globosa (5) and P. flavescens (1). Likewise, specimens of C. neoformans presented molecular type VNI and molecular type VNII; for C. gattii the molecular types found were VGIII and one VGIV by URA5-RFLP but VGIII by MALDI-TOF and sequencing of the ITS and LSU. Therefore, it can be concluded that the species of Cryptococcus, Naganishia and Papiliotrema genera, are present in the environment of Boyacá, and show a predilection for climate conditions that are typical of this region.
