Background: Dermatophytosis represents one of the common infectious diseases worldwide and it is a major public health problem around the globe. The disease causes considerable morbidity and still continues to increase especially in developing countries. Objective: This study was undertaken to determine the prevalence of dermatophytes and the spectrum of fungal agents in patients attending Rank Higher Specialized Dermatology Clinic. Methods: A cross-sectional study has been conducted, in which 318 samples from 318 suspected patients were collected. Samples include hair, nail, and skin. A portion of each sample was examined microscopically and the remaining portion of each sample was cultured onto plates of Sabouraud’s dextrose agar containing chloramphenicol with and without cycloheximide. Isolates were identified by studying the macroscopic and microscopic characteristics of the colonies. Results: Tinea capitis was the predominant clinical manifestation accounting for 53.4% of the cases. Patients with age group 1– 14 years were more affected. Of 318 samples, fungi were detected in 133 (54.4%) by direct wet mount while 148/315 (46.5%) of them were culture positive. From these 72/148 (46.8%) were dermatophytes. T. tonsurans was the most common pathogen in tinea capitis, whereas T. mentagrophytes was the most common pathogen in tinea corporis. Among dermatophyte isolates, T. tonsurans 29/72 (40.2%) was the most common cause of infection. Among non-dermatophyte molds, Cladosporium spp. 21/63 (33.3%) was predominant isolate followed by Neoscytalidim dimidatum 11/63 (17.4%) and Alternaria spp. 9/63 (14.2%), respectively. Yeasts also account for 13 (8.7%) of the total suspects of dermatophytosis. Conclusion: In this study, the prevalence of dermatophytes was higher in tinea capitis 46/72 (63.8%) and T. tonsurans 29/72 (40%) was the dominant-isolated dermatophyte. Recovery of a large number of dermatophytes and non-dermatophyte fungi in our study showed that non-dermatophyte fungi are emerging as important causes of dermatophytosis warranting further intensive epidemiological studies that have public health significance are needed. Keywords: dermatophytes, non-dermatophytes, dermatophytosis, fungi, infection
Urbanization is increasing worldwide and is regarded a major threat to biodiversity in forests. As consequences of intensive human use, the vegetation structure of naturally growing urban forests and their amount of deadwood can be reduced. Deadwood is an essential resource for various saproxylic insects and fungi. We assessed the effects of urbanization and forest characteristics on saproxylic insects and fungi. We exposed standardized bundles consisting of each three freshly cut beech and oak branches in 25 forests along a rural-urban gradient in Basel (Switzerland). After an exposure of 8 months, we extracted the saproxylic insects for 10 months using an emergence trap for each bundle. We used drilling chips from each branch to determine fungal operational taxonomic units (OTUs). In all, 193,534 insect individuals emerged from the experimental bundles. Our study showed that the abundance of total saproxylic insects, bark beetles, longhorn beetles, total flies, moths, and ichneumonid wasps decreased with increasing degree of urbanization, but not their species richness. However, the taxonomic composition of all insect groups combined was altered by wood moisture of branches and that of saproxylic beetles was influenced by the degree of urbanization. Unexpectedly, forest size and local forest characteristics had a minor effect on saproxylic insects. ITS (internal transcribed spacer of rDNA) analysis with fungal specific primers revealed a total of 97 fungal OTUs on the bundles. The number of total fungal OTUs decreased with increasing degree of urbanization and was affected by the volume of naturally occurring fine woody debris. The composition of fungal OTUs was altered by the degree of urbanization and pH of the branch wood. As a consequence of the altered compositions of saproxylics, the association between total saproxylic insects and fungi changed along the rural-urban gradient. Our study shows that urbanization can negatively impact saproxylic insects and fungi.
Edibles are the source of life, providing the body with essential nutrients to maintain physiological functions. As far back as ancient times, people have used edibles to treat diseases, also known as dietary therapy. Edible fungi, as favored foods, not only have rich nutritional value but also have various pharmacological effects and have a significant role in the treatment of insomnia. This review delves into the potential mechanisms of edible fungi in the sedative-hypnotic effect, focusing on several species of edible fungi with remarkable sleep-improving effects including <i>Cordyceps militaris</i>, <i>Ganoderma lucidum</i>, <i>Poria cocos</i>, <i>Hericium erinaceus</i> and <i>Armillaria mellea</i>. The modulatory effects of active ingredients, including polysaccharides, triterpenes, and nucleosides, found in edible fungi, on the neurotransmitter system, antioxidant, anti-inflammatory pathways, gut microflora homeostasis, and immune function are reviewed. For instance, <i>C. militaris</i> reduces excitatory neurotransmitters like glutamate and acetylcholine. <i>P. cocos</i> polysaccharides regulate gut flora to positively influence sleep. <i>G. lucidum</i> enhances <i>γ</i>-aminobutyric acid (GABA) and serotonin expression, boosts antioxidant activity, and reduces inflammatory mediators, collectively improving sleep quality. In addition to the aforementioned improvements on insomnia by the active components in edible fungi, they also exert sedative and hypnotic effects through other mechanisms. This review summarizes the sedative and hypnotic mechanisms of these edible fungi, providing new perspectives and methodologies for the research on the sedative and hypnotic activity of Changbai Mountain <i>C. militaris</i> and other edible fungi, which is conducive to exploring and utilizing more edible fungal resources from the Changbai Mountain with such activities.
Endophytic fungi have emerged as a significant source of natural products with remarkable bioactivities. Recent research has identified numerous antioxidant molecules among the secondary metabolites of endophytic fungi. These organisms, whether unicellular or micro-multicellular, offer the potential for genetic manipulation to enhance the production of these valuable antioxidant compounds, which hold promise for promoting health, vitality, and various biotechnological applications. In this study, we provide a critical review of methods for extracting, purifying, characterizing, and estimating the total antioxidant capacity (TAC) of endophytic fungi metabolites. While many endophytes produce metabolites similar to those found in plants with established symbiotic associations, we also highlight the existence of novel metabolites with potential scientific interest. Additionally, we discuss how advancements in nanotechnology have opened new avenues for exploring nanoformulations of endophytic metabolites in future studies, offering opportunities for diverse biological and industrial applications.
Development of biological control for plant diseases is accepted as a durable and environmentally friendly alternative for agrochemicals. Arbuscular mycorrhizal fungi (AMF), which form symbiotic associations with root systems of most agricultural, horticultural and hardwood crop species, have been suggested as widespread potential bioprotective agents. In the present study the ability of two AMF (Glomus mosseae and Glomus intraradices) to induce local or systemic resistance to Phytophthora parasitica in tomato roots have been compared using a split root experimental system. Glomus mosseae was effective in reducing disease symptoms produced by P. parasitica infection, and evidence points to a combination of local and systemic mechanisms being responsible for this bioprotector effect. The biochemical analysis of different plant defence-related enzymes showed a local induction of mycorrhiza-related new isoforms of the hydrolytic enzymes chitinase, chitosanase and beta-1,3-glucanase, as well as superoxide dismutase, an enzyme which is involved in cell protection against oxidative stress. Systemic alterations of the activity of some of the constitutive isoforms were also observed in non-mycorrhizal roots of mycorrhizal plants. Studies on the lytic activity against Phytophthora cell wall of root protein extracts also corroborated a systemic effect of mycorrhizal symbiosis on tomato resistance to Phytophthora.
Endophytes fungi are applied as favorable safe antifungal agents as well as natural bioactive compounds reservoir. In the current study, the inhibitory effect of endophytic fungus was explained by direct antifungal activity against fungi causing mucormycosis, ultrastructural, and determination of active compounds in fungal extract. Endophytic Aspergillus terreus was isolated from healthy Moringa oleifera leaves and identified morphologically and genetically, and was recorded in gene bank with accession number MW444551.1. Phytochemical analysis and gas chromatography-mass spectroscopy (GC-MS) of ethyl acetate crude extract (EACE) of A. terreus were performed. GC-MS results of EACE of A. terreus revealed that fungal extract contains 16 major bioactive compounds with extensive pharmaceutical activities. Furthermore, EACE of A. terreus revealed a promising antifungal activity against fungi causing mucormycosis as Rhizopus oryzae, Mucor racemosus, and Syncephalastrum racemosum, where inhibition zones of EACE (10 mg/ml) were 20, 37, and 18 mm, respectively. Minimum inhibitory concentration (MIC) of EACE was 0.3125 toward M. racemosus, while 1.25 and 2.5 mg/ml against R. oryzae and S. racemosum, respectively. In the same context, treated R. oryzae, M. racemosus, and S. racemosum with EACE of A. terreus revealed elevation of membrane lipid peroxidation which approves membrane leakage. Furthermore, ultrastructure changes were observed which established alteration in both sporangium and hyphal structures; cell membrane and cytoplasm leakage. In conclusion, endophytic A. terreus has an outstanding antifungal activity against fungi causing mucormycosis.
Monilinia species are among the most devastating fungi worldwide as they cause brown rot and blossom blight on fruit trees. To understand the molecular bases of their pathogenic lifestyles, we compared the newly assembled genomes of single strains of Monilinia fructicola , M. fructigena and M. laxa , with those of Botrytis cinerea and Sclerotinia sclerotiorum , as the closest species within Sclerotiniaceae . Phylogenomic analysis of orthologous proteins and syntenic investigation suggest that M. laxa is closer to M. fructigena than M. fructicola , and is closest to the other investigated Sclerotiniaceae species. This indicates that M. laxa was the earliest result of the speciation process. Distinct evolutionary profiles were observed for transposable elements (TEs). M. fructicola and M. laxa showed older bursts of TE insertions, which were affected (mainly in M. fructicola ) by repeat-induced point (RIP) mutation gene silencing mechanisms. These suggested frequent occurrence of the sexual process in M. fructicola . More recent TE expansion linked with low RIP action was observed in M. fructigena , with very little in S. sclerotiorum and B. cinerea . The detection of active non-syntenic TEs is indicative of horizontal gene transfer and has resulted in alterations in specific gene functions. Analysis of candidate effectors, biosynthetic gene clusters for secondary metabolites and carbohydrate-active enzymes, indicated that Monilinia genus has multiple virulence mechanisms to infect host plants, including toxins, cell-death elicitor, putative virulence factors and cell-wall-degrading enzymes. Some species-specific pathogenic factors might explain differences in terms of host plant and organ preferences between M. fructigena and the other two Monilinia species.
Edible fungi are large fungi with high added value that can be utilized as resources. They are rich in high-quality protein, carbohydrate, various vitamins, mineral elements and other nutrients, and are characterized by high protein, low sugar, low fat and low cholesterol. In addition, edible fungi contain a variety of bioactive substances, such as polysaccharides, dietary fiber, steroids, polyphenols, and most of these compounds have antioxidant, anti-tumor and other physiological functions. This review comprehensively discusses the bioactive components and functional characteristics of edible fungi (such as antioxidant, anti-aging, hypolipidemic activities, etc.). Then the recent developments and prospect in the high-valued utilization of edible fungi are discussed and summarized. The objective of this review is to improve the understanding of health-promoting properties of edible fungi, and provide reference for the industrial production of edible fungi-based health products.
We studied the transport of 15 N from a soil compartment separated from a plant root compartment by a hydrophobic polytetrafluoroethylene (PTFE) membrane to plants in the presence and absence of arbuscular mycorrhizal fungi (AMF). We have previously shown that this type of membrane efficiently inhibits mass flow and diffusion of mobile ions in the soil solution in an abiotic system, but can be penetrated by the hyphae of mycorrhizal fungi. Mycorrhizal tomatoes ( Lycopersicon esculentum ) colonized by Glomus mosseae were grown at two N fertilizer concentrations in a root compartment. A PTFE membrane was placed between the root compartment and an adjoining soil compartment that was inaccessible to the roots but accessible to the AMF hyphae (hyphal compartment). Additional N was applied to the hyphal compartment using uniformly 15 N‐labelled NH 4 NO 3 . There was a flux of 15 N from the hyphal compartment to the plants even in the absence of mycorrhizal fungi. However, this flux was much higher in mycorrhizal plants, which had much higher N concentrations in their shoots and roots than did the non‐mycorrhizal control plants. This was particularly apparent when the root compartment had a low N fertilizer concentration. Of the total N content of mycorrhizal plants, c . 42 and 24% at the low and high N fertilizer concentrations, respectively, were estimated to originate from the hyphal compartment by transport through AMF hyphae. In the presence of mycorrhizal fungi, the flux of 15 N was about three times higher than in their absence. The results show that AMF can access a soil compartment separated by a PTFE membrane, and can contribute substantially to N uptake by plants.
Many species of fungi including lichenized fungi (lichens) and algae have the ability to biosynthesize biologically active compounds. They produce, among others, polysaccharides with anticancer and immunostimulatory properties: (1) Background: This paper presents the characteristics of the most important bioactive compounds produced by fungi and algae; (2) Methods: Based on the example of the selected species of mushrooms, lichens and algae, the therapeutic properties of the secondary metabolites that they produce and the possibilities of their use are presented; (3) Results: The importance of fungi, especially large-fruited mushrooms, lichens and algae, in nature and human life is discussed, in particular, with regard to their use in the pharmaceutical industry and their nutritional value; (4) Conclusions: The natural organisms, such as fungi, lichenized fungi and algae, could be used as supplementary medicine, in the form of pharmaceutical preparations and food sources. Further advanced studies are required on the pharmacological properties and bioactive compounds of these organisms.
The impact of colonization by the arbuscular mycorrhizal fungus (AMF) Glomus mosseae on tomato root necrosis caused by the soil‐borne pathogen Phytophthora parasitica was investigated. Studies in situ permitted infection loci to be identified and the effects of the AMF on numbers to be elucidated. Effects were significant and, 7 and 16 days after inoculation with zoospores of the pathogen, roots of plants colonized by the AMF had 39% and 30%, respectively, fewer infection loci than those that were not. Concurrent studies of the rate of spread of necrosis within roots showed no changes caused by the AMF. At harvest, 26 days following inoculation with the pathogen, 61% of roots of noncolonized plants were necrotic compared with only 31% in AMF‐colonized plants. It is concluded that effects on numbers of infection loci are one mechanism via which AMF achieve biocontrol of this pathogen in tomato. Measures of the effects of the AMF on root system architecture suggest that no significant changes occur and are thus not the reasons for the reduction in infection loci. The implications of these data for agricultural practice and biocontrol research are discussed.
Endophytic fungi residing in medicinal plants have the ability to produce the same pharmacologic bioactive secondary metabolites as their host medicinal plants, which have been used for thousands of years in traditional medicine and still are used for their health benefits. Nowadays, medicinal plants are quarrying for isolation of plant-derived drugs as they are very effective and have reasonably less or no side effects. However, the natural resources of ethanomedicinal plants are gradually exhausted and access to plant bioactive compounds is challenged by the low levels at which these products accumulate in native medicinal plants. For example, to meet the market demands of 3 Kg per year of Vinca alkaloids, powerful plant-derived anticancer drugs, 1.5x106 Kg dry leaves are required. In this regard, this review articles aims to highlight the fact that endophytic fungi residing in medicinal plants are capable to biosynthesize pharmacologically active secondary metabolites as antioxidant identical to those produced by their host medicinal plant. Furthermore, the evolutionary origin of the genes involved in these metabolic pathways as well as the approaches designed to enhance the production of these secondary metabolites by the isolated endophytic fungi medicinal plant have a lot of type of antioxidant mostly polyphenols, flavonoids which exhibit high antioxidants bioactivity. In addition to aforementioned reasons, this article also will shed the light on the efforts of Abdel-Azeem and his co-workers at Botany department, Faculty of Science, Suez Canal University and their continuing search for biologically active natural products from Egyptian endophytic fungi hosted medicinal plants in Saint Katherine Protectorate, Arid Sinai, Egypt.
Communities of arbuscular mycorrhizal fungi (AMF) in five agricultural field sites of different management intensities were studied. Variable regions of the ribosomal RNA genes were used to detect and identify AMF directly within colonized roots. Roots from a continuous maize monoculture showed low AMF diversity, in agreement with previous reports on molecular diversity of AMF in agricultural soils. In contrast, a substantially higher diversity of AMF was found throughout the long term 'DOK' field experiment, where organic and conventional agricultural practices have been compared side by side since 1978. In this experiment, a 7-year crop rotation is performed under lower levels of inorganic fertilizer input and chemical pest control. These results are in good agreement with analyses of the spore community previously conducted in these field sites. In a third site, an organically managed leek field with soil of very high phosphate content reflecting the highly intensive conventional field history and intensive tillage, we detected a low-diversity community comparable to the maize monoculture. In addition to fungi from Glomus group A, which have previously been reported to dominate arable soils, we regularly found members of the genera Scutellospora, Paraglomus and Acaulospora. The genus Acaulospora was shown to occur more frequently early in the growing season, suggesting that the life history strategy of AMF may influence the active community at a given time. These data show that the diversity of AMF is not always low in arable soils. Furthermore, low-input agriculture involving crop rotation may provide better conditions to preserve AMF diversity, by preventing the selection for the few AMF taxa tolerating high nutrient levels.
The gut mycobiome significantly affects host health and immunity. However, most studies have focused on symbiotic bacteria in the gut microbiome, whereas less attention has been given to symbiotic fungi. Although fungi constitute only 0.01%-0.1% of the gut microbiome, their larger size and unique immunoregulatory functions make them significant. Factors like diet, antimicrobials use, and age can disrupt the fungal community, leading to dysbiosis. Fungal-bacterial-host immune interactions are critical in maintaining gut homeostasis, with fungi playing a role in mediating immune responses such as Th17 cell activation. This review highlights methods for studying gut fungi, the composition and influencing factors of the gut mycobiome, and its potential in therapeutic interventions for intestinal and hepatic diseases. We aim to provide new insights into the underexplored role of gut fungi in human health.
Arbuscular mycorrhiza fungi (AMF) are beneficial soil fungi that can promote the growth of their host plants. Accurate quantification of AMF in plant roots is important because the level of colonization is often indicative of the activity of these fungi. Root colonization is traditionally measured with microscopy methods which visualize fungal structures inside roots. Microscopy methods are labor-intensive, and results depend on the observer. In this study, we present a relative qPCR method to quantify AMF in which we normalized the AMF qPCR signal relative to a plant gene. First, we validated the primer pair AMG1F and AM1 in silico, and we show that these primers cover most AMF species present in plant roots without amplifying host DNA. Next, we compared the relative qPCR method with traditional microscopy based on a greenhouse experiment with Petunia plants that ranged from very high to very low levels of AMF root colonization. Finally, by sequencing the qPCR amplicons with MiSeq, we experimentally confirmed that the primer pair excludes plant DNA while amplifying mostly AMF. Most importantly, our relative qPCR approach was capable of discriminating quantitative differences in AMF root colonization and it strongly correlated (Spearman Rho = 0.875) with quantifications by traditional microscopy. Finally, we provide a balanced discussion about the strengths and weaknesses of microscopy and qPCR methods. In conclusion, the tested approach of relative qPCR presents a reliable alternative method to quantify AMF root colonization that is less operator-dependent than traditional microscopy and offers scalability to high-throughput analyses.
Fungi are amongst the most abundant and diverse organisms. Despite being widely known for their adverse role in food spoilage or as pathogens for humans, animals, or plants, they also present several beneficial effects. Fungi contribute to human well-being due to their role as decomposers, degrading decay matter into smaller molecules which can be easily used by other ecosystem members. These organisms can produce medicinal compounds or modulate protective immune responses in human intestine. Fungi intervene in diverse food processes or act as a food supply. Due to fungal diversity, the unequivocal identification of these organisms is crucial to increasing their practical applications and decreasing their adverse effects. The process of identification could be achieved through the integral sequencing of fungi genomes. However, this procedure would be time-consuming and rather cost-inefficient. Therefore, several molecular markers have been developed to overcome these limitations. The chronology of DNA-based molecular markers development can be divided into three main steps: (1) prior to the development of the PCR technique (RFLP); (2) after the development of the PCR technique (RAPD, AFLP, ISSR, VNTR, SNP, InDels, and DNA barcoding); (3) after the development of the massive parallel sequencing technique (Metabarcoding and WGS). Therefore, the present review covers an overview of the most recently developed molecular markers used for fungal detection and identification.
How community-level specialization differs among groups of organisms, and changes along environmental gradients, is fundamental to understanding the mechanisms influencing ecological communities. In this paper, we investigate the specialization of root-associated fungi for plant species, asking whether the level of specialization varies with elevation. For this, we applied DNA barcoding based on the ITS region to root samples of five plant species equivalently sampled along an elevational gradient at a high arctic site. To assess whether the level of specialization changed with elevation and whether the observed patterns varied between mycorrhizal and endophytic fungi, we applied a joint species distribution modeling approach. Our results show that host plant specialization is not environmentally constrained in arctic root-associated fungal communities, since there was no evidence for changing specialization with elevation, even if the composition of root-associated fungal communities changed substantially. However, the level of specialization for particular plant species differed among fungal groups, root-associated endophytic fungal communities being highly specialized on particular host species, and mycorrhizal fungi showing almost no signs of specialization. Our results suggest that plant identity affects associated mycorrhizal and endophytic fungi differently, highlighting the need of considering both endophytic and mycorrhizal fungi when studying specialization in root-associated fungal communities.
Plant growth is limited in arid and/or contaminated sites due to the adverse conditions coming from heavy metal (HM) contamination and/or water stress.Moreover, soils from these areas are generally characterised by poor soil structure, low water-holding capacity, lack of organic matter and nutrient deficiency.In order to carry out a successful re-afforestation, it is necessary to improve soil quality and the ability of plants species to resist this harsh environment.The symbiosis with arbuscular mycorrhizal (AM) fungi has been proposed as one of the mechanisms of plant heavy metal tolerance and water stress avoidance.On the other hand, addition of organic amendments to the soil can reverse degradation of soil properties.Agro-waste residues such as dry olive cake (DOC) and sugar beet waste (SB) supplemented with rock phosphate (RP) can be used as organic amendments after fermentation by Aspergillus niger.The application of A. niger-treated DOC and/or SB to semi-arid soils and/or HM-contaminated soils increased aggregate stability, soil enzymatic activities, water soluble C and water soluble carbohydrates as well as nutrient availability, especially P. AM inoculation, using adapted endophytes, was more efficient with respect to increasing plant nutrition and growth as well as plant tolerance to drought or HM-stress conditions.The combined treatments involving mycorrhiza fungi inoculation and addition of the amendments into the soil can be proposed as a successful revegetation strategy for plant performance in P-deficient soils under semiarid Mediterranean conditions.The beneficial effectiveness of this symbiosis with suitable AM fungi in A. niger-treated agro-waste residue-amended soil can also be regarded as a successful biotechnological tool for reclamation of HM-contaminated soils.
The improvement of the nutritional quality of lettuce by its association with arbuscular mycorrhizal fungi (AMF) has been recently reported in a previous study. The aim of this research was to evaluate if the fertilization with three P sources differing in water solubility affects the effectiveness of AMF for improving lettuce growth and nutritional quality. The application of either water-soluble P sources (Hewitt's solution and single superphosphate) or the water-insoluble (WI) fraction of a "rhizosphere-controlled fertilizer" did not exert negative effects on the establishment of the mycorrhizal symbiosis. AMF improved lettuce growth and nutritional quality. Nevertheless, the effect was dependent on the source of P and cultivar. Batavia Rubia Munguía (green cultivar) benefited more than Maravilla de Verano (red cultivar) in terms of mineral nutrients, total soluble sugars, and ascorbate contents. The association of lettuce with AMF resulted in greater quantities of anthocyanins in plants fertilized with WI, carotenoids when plants received either Hewitt's solution or WI, and phenolics regardless of the P fertilizer applied.
INTRODUCTION: The aim of the study was the evaluation of panfungal PCR protocols with subsequent sequence analysis for the diagnostic identification of invasive mycoses in formalin-fixed, paraffin-embedded tissue samples with rare tropical mycoses. MATERIALS AND METHODS: Five different previously described panfungal PCR/sequencing protocols targeting 18S and 28S ribosomal RNA gene fragments as well as internal transcribed spacer 1 and 2 fragments were evaluated with a collection of 17 formalin-fixed, paraffin-embedded tissue samples of patients with rare and/or tropical invasive mycoses, comprising chromoblastomycosis, coccidioidomycosis, cryptococcosis, histoplasmosis, mucormycosis, mycetoma/maduromycosis, and rhinosporidiosis, in a proof-of-principle analysis. RESULTS: The primers of the panfungal PCRs readily and predominantly reacted with contaminating environmental fungi that had deposited on the paraffin blocks. Altogether three sequence results of histoplasmosis and mycetoma samples that matched the histological assessment were associated with sample age <10 years and virtually without PCR inhibition. CONCLUSIONS: The high risk of amplifying environmental contaminants severely reduces the usefulness of the assessed panfungal PCR/sequencing protocols for the identification of rare and/or tropical mycoses in stored formalin-fixed, paraffin-embedded tissues. Histological assessment remains valuable for such indications if cultural differentiation is impossible from inactivated sample material.