Review and Current Perspectives on DNA Topoisomerase I and II Enzymes of Fungi as Study Models for the Development of New Antifungal Drugs.

Fungal infections represent a growing public health problem, mainly stemming from two phenomena. Firstly, certain diseases (e.g., AIDS and COVID-19) have emerged that weaken the immune system, leaving patients susceptible to opportunistic pathogens. Secondly, an increasing number of pathogenic fungi are developing multi-drug resistance. Consequently, there is a need for new antifungal drugs with novel therapeutic targets, such as type I and II DNA topoisomerase enzymes of fungal organisms. This contribution summarizes the available information in the literature on the biology, topology, structural characteristics, and genes of topoisomerase (Topo) I and II enzymes in humans, two other mammals, and 29 fungi (including Basidiomycetes and Ascomycetes). The evidence of these enzymes as alternative targets for antifungal therapy is presented, as is a broad spectrum of Topo I and II inhibitors. Research has revealed the genes responsible for encoding the Topo I and II enzymes of fungal organisms and the amino acid residues and nucleotide residues at the active sites of the enzymes that are involved in the binding mode of topoisomerase inhibitors. Such residues are highly conserved. According to molecular docking studies, antifungal Topo I and II inhibitors have good affinity for the active site of the respective enzymes. The evidence presented in the current review supports the proposal of the suitability of Topo I and II enzymes as molecular targets for new antifungal drugs, which may be used in the future in combined therapies for the treatment of infections caused by fungal organisms.

Inhibition of Aspergillus spp. growth and ochratoxin A production in Conilon and Arabica coffees based-medium by Saccharomyces cerevisiae.

Saccharomyces cerevisiae CCMA 0159 is reported as a promising biocontrol agent against ochratoxin A (OTA)-producing fungi in coffee. Coffea arabica and Coffea canephora (var. Conilon or Robusta) are the most widely consumed coffee species around the world, cultivated in tropical and subtropical regions, each exhibiting distinct physicochemical and sensory characteristics. The objective of this study was to compare the growth and OTA production by Aspergillus carbonarius, A. ochraceus, and A. westerdijkiae in C. arabica and C. canephora, along with assessing the efficiency of S. cerevisiae CCMA 0159 in biocontrolling ochratoxigenic fungi in both coffee varieties. A. carbonarius exhibited a higher growth rate and OTA production in both coffee varieties, with C. canephora showing particular susceptibility. Conversely, A. ochraceus and A. westerdijkiae demonstrated lower growth and OTA production. S. cerevisiae was effective in biocontrolling the fungal isolates, inhibiting over 80 % of A. carbonarius growth in both coffee varieties. Among the mechanisms of action of the biological control agent, the production of volatile organic compounds stands out. The results of this study confirm the significant potential of S. cerevisiae CCMA 0159 as a biocontrol agent against Aspergillus for application in coffee-producing areas.

Enhanced degradation of phototreated recycled and unused low-density polyethylene films by Pleurotus ostreatus.

Polyethylene, one of the most used petroleum-derived polymers, causes serious environmental pollution. The ability of Pleurotus ostreatus to degrade UV-treated and untreated recycled and unused (new) low-density polyethylene (LDPE) films was studied. We determined the fungal biomass production, enzyme production, and enzyme yield. Changes in the chemical structure and surface morphology of the LDPE after fungal growth were analyzed using FTIR spectroscopy and SEM. Functional group indices and contact angles were also evaluated. In general, the highest Lac (6013 U/L), LiP (2432 U/L), MnP (995 U/L) and UP (6671 U/L) activities were observed in irradiated recycled LDPE (IrRPE). The contact angle of all samples was negatively correlated with fermentation time; the smaller the contact angle, the longer the fermentation time, indicating effective biodegradation. The IrRPE samples exhibited the smallest contact angle (49°) at 4 weeks, and the samples were fragmented (into two pieces) at 5 weeks. This fungus could degrade unused (new) LDPE significantly within 6 weeks. The biodegradation of LDPE proceeded faster in recycled than in unused samples, which can be enhanced by exposing LDPE to UV radiation. Enzymatic production during fungal growth suggest that LDPE degradation is initiated by laccase (Lac) followed by lignin peroxidase (LiP), whereas manganese peroxidase (MnP) and unspecific peroxygenase (UP) are involved in the final degradation process. This is the first experimental study on the fungal growth and its main enzymes involved in LDPE biodegradation. This fungus has great promise as a safe, efficient, and environmentally friendly organism capable of degrading LDPE.

Isolation, genome analysis and tissue localization of Ceratobasidium theobromae, a new encounter pathogen of cassava in Southeast Asia.

In Southeast Asia (SEA) fastidious fungi of the Ceratobasidium genus are associated with proliferation of sprouts and vascular necrosis in cacao and cassava, crops that were introduced from the tropical Americas to this region. Here, we report the isolation and in vitro culture of a Ceratobasidium sp. isolated from cassava with symptoms of witches' broom disease (CWBD), a devastating disease of this crop in SEA. The genome characterization using a hybrid assembly strategy identifies the fungus as an isolate of the species C. theobromae, the causal agent of vascular streak dieback of cacao in SEA. Both fungi have a genome size > 31 Mb (G+C content 49%), share > 98% nucleotide identity of the Internal Transcribed Spacer (ITS) and > 94% in genes used for species-level identification. Using RNAscope® we traced the pathogen and confirmed its irregular distribution in the xylem and epidermis along the cassava stem, which explains the obtention of healthy planting material from symptom-free parts of a diseased plant. These results are essential for understanding the epidemiology of CWBD, as a basis for disease management including measures to prevent further spread and minimize the risk of introducing C. theobromae via long-distance movement of cassava materials to Africa and the Americas.

Interaction of Capnodium alfenasii with extrafloral nectaries of Azadirachta indica.

Sooty moulds are saprophytic epiphytic fungi that grow mostly on insect secretions, but they can also be associated with plant secretions. In this study, we aimed to describe de interaction of Capnodium alfenasii sooty mould with the extrafloral shoot nectaries of Azadirachta indica. Anatomical and histochemical studies were carried out on serial sections of extrafloral shoot nectaries of A. indica without and with C. alfenasii infestation. The total soluble sugar content of the secreted nectar was determined, and the conidial germination of the fungus in distilled water and in dextrose and nectar solutions was evaluated. The shoot nectaries of A. indica are elongated structures that occur in pairs near the base of the petiole. The exuded nectar contains an average of 534.8 µg of total soluble sugars per µL of nectar and provides ideal conditions for conidial germination and fungal growth. C. alfenasii hyphae grow on the nectary, penetrate through breaks in the cuticle, travel under the cuticle and penetrate the secretory tissue by inter- and intracellular routes. The present report is the first to describe the interaction of C. alfenasii with the A. indica nectary, including the penetration of hyphae into nectariferous tissues and the plant defence mechanisms.

A Countermeasure Strategy against Peramine Developed by Chilesia rudis in the Endophyte-Ryegrass-Herbivore Model.

Exploitation of the symbiotic relationship between endophytic fungi and ryegrass is a crucial technique for reducing the incidence of insect pests. This is primarily due to the production of alkaloids, such as peramine, by the fungi. This alkaloid has been reported as both a deterrent and toxic to a variety of insects. However, insects have developed various strategies to counteract plant defenses. One of the most studied methods is their ability to sequester toxic compounds from plants. In this study, we examined the feeding preferences and adaptation to peramine in Chilesia rudis, a native Chilean larva. Using a no-choice assay, we assessed larval feeding preferences and mass gain on seven experimental lines and two commercial cultivars of endophyte-infected and non-infected ryegrass. Pupal development time and adult performance were evaluated post-assay. Additionally, we measured peramine content in larval carcasses, feces, and ryegrass leaves. Jumbo was the most preferred cultivar with 32 mm2 of leaf tissues consumed. The longest pupal development time was observed in L161 and ALTO AR1, both at 28 days. Wing length in adults was greatest in the Jumbo and L163 cultivars, measuring 1.25 cm and 1.32 cm, respectively. Peramine concentrations were detected in the bodies of C. rudis. In conclusion, this larva can adapt to endophyte-infected ryegrass and develop counter-adaptation mechanisms to mitigate the effects of peramine.

Aspergillus brasiliensis E_15.1: A Novel Thermophilic Endophyte from a Volcanic Crater Unveiled through Comprehensive Genome-Wide, Phenotypic Analysis, and Plant Growth-Promoting Trails.

Thermophilic fungi have been seldom studied despite the fact that they can contribute to understanding ecological mechanisms of adaptation in diverse environments and have attractive toolboxes with a wide range of biotechnological applications. This work describes for the first time an endophytic and thermophilic strain of Aspergillus brasiliensis that was isolated in the crater of the active volcano "El Chichonal" in Mexico. This strain was capable of surviving in soil with a temperature of 60 °C and a pH of neutral acidity, which preluded a high thermostability and a potential in industrial application. The complete genome of A. brasiliensis E_15.1 was sequenced and assembled in 37 Mb of genomic DNA. We performed a comprehensive phylogenomic analysis for the precise taxonomic identification of this species as a novel strain of Aspergillus brasiliensis. Likewise, the predicted coding sequences were classified according to various functions including Carbohydrate-Active Enzymes (CAZymes), biosynthetic gene clusters of secondary metabolites (BGCs), and metabolic pathways associated with plant growth promotion. A. brasiliensis E_15.1 was found to degrade chitin, chitooligosaccharides, xylan, and cellulose. The genes to biosynthesize clavaric acid (a triterpene with antitumor activity) were found, thus probably having antitumor activity. In addition to the genomic analysis, a set of enzymatic assays confirmed the thermostability of extracellular xylanases and cellulases of A. brasiliensis E_15.1. The enzymatic repertoire of A. brasiliensis E_15.1 suggests that A. brasiliensis E_15.1 has a high potential for industrial application due to its thermostability and can promote plant growth at high temperatures. Finally, this strain constitutes an interesting source of terpenoids with pharmacological activity.

Culture-dependent methods revel the diversity of endophytic fungi of Psidium cattleianum leaves (Myrtales: Myrtaceae).

Endophytic fungi are a diverse group of microorganisms that reside within plant tissues and play a crucial ecological role in the development of their hosts. Psidium cattleianum (Myrtales: Myrtaceae: 'Cattley guava') is a Brazilian native species with economic potential due to the diverse applications of its fruits, wood, and essential oils. Despite their significance, the diversity of endophytic fungi associated with P. cattleianum remains unexplored. Here, we investigated the diversity of endophytic fungi in the leaves of this plant using cultivation-dependent isolation methods, analysis of the macroscopic characters of the isolates, and phylogenetic analyses employing the ITS barcode marker. A total of 396 isolates, classified into 25 fungal taxa, were obtained, namely, Alternaria, Aspergillus, Cladosporium, Colletotrichum, Coprinellus, Coriolopsis, Diaporthe, Induratia, Mycosphaerella, Muyocoprom, Myrmecridium, Neofusicoccum, Pantospora, Paracamarosporium, Parapallidocercospora, Paraphaeosphaeria, Penicillium, Perenniporia, Phaeophleospora, Phyllosticta, Pseudofusicoccum, Talaromyces, Xylaria, Sordariomycetes, and Xylariomycetes. Our findings reveal a significant diversity of fungi associated with P. cattleianum leaves; however, our study suggests an even greater diversity of fungi associated with this plant species. Interestingly, although P. cattleianum shares endophytic fungi with other plants in the Myrtaceae family, this plant species harbors a unique fungal community. This distinction is evidenced by certain fungal genera and seven potentially new phylogenetic species, isolated in this study.

Intra-Phenotypic and -Genotypic Variations of Beauveria bassiana (Bals.) Vuill. Strains Infecting Aedes aegypti L. Adults.

Beauveria bassiana has potential for Aedes aegypti biological control. However, its efficacy depends on the strain's geographic location, host susceptibility, and virulence. The present study aimed to evaluate the effectiveness of B. bassiana strain BBPTG4 conidia in controlling Ae. aegypti adults and its detection via introns profile on exposed mosquito corpses. Morphologic characteristics among strains were highly similar. Comprehensive testing of these strains demonstrated that BBPT4 exhibited the ideal biological activity for Ae. aegypti control, with a median lethal time (TL50) of 7.5 d compared to ~3 d and ~10 d for BB01 and BB37 strains, respectively. Infected mosquitoes died after GHA and BBPTG4 exposure, and corpses were analyzed for infecting strains detection. Differences among the seven evaluated strains were determined, assessing five different insertion group I intron profiles in BBTG4, BB01, GHA, BB37, and BB02 strains. Mosquitoes infected by BBPTG4 and non-exposed (negative control) intron profiles were obtained. We detected the presence of introns in the BBPTG4 strain, which were not present in non-exposed mosquitoes. In conclusion, B. bassiana strains showed similarities in terms of their cultural and microscopic morphological characteristics and biologicals virulence level, but different intron profiles. BBPTG4 strain-infected Ae. aegypti adult corpses, showing specific amplicons, enabled us to identify B. bassiana at the strain level among infected mosquitoes. However, monitoring and detection of field-infected insects is essential for further verification.

Investigation into the effect of the culture conditions and optimization on limonene-1,2-diol production from the biotransformation of limonene using Pestalotiopsis mangiferae LaBMicrA-505.

Different bioproducts can be obtained by changing operative condition of biotechnological process, and this bioprocess aspect is a significant approach to be adopted on industrial scale leading to the creation of new natural aroma. Thus, this study aimed to investigate the culture conditions and optimization of the biotransformation of limonene into limonene-1,2-diol using Pestalotiopsis mangiferae LaBMicrA-505 obtained from the Brazilian Amazon. The study started with the investigation of the establishment of culture, followed by optimization of the conditions for biotransformation of R-(+)-limonene to limonene-1,2-diol, using shake flasks. The fresh biomass of P. mangiferae LaBMicrA-505 obtained in liquid media supplemented with yeast-malt extract under with 72 h (stationary phase) performed better diol productivity when compared to other biomasses. Finally, in the modeling of contour plots and surface responses of a central composite design, the use of 4 g l- 1 biomass, 2% of the substrate at 24 °C, 120 rpm, and pH of 6.0 could maximize the production of limonene-1,2-diol, accumulated up to 98.34 ± 1.53% after 96 h of reaction. This study contributed to identified operational condition for the R-(+)-limonene bioconversion scale-up. The endophytic fungus P. mangiferae LaBMicrA-505 proved to be a potent biocatalyst to biotechnologically produce limonene-1,2-diol, an aroma compounds with interesting bioactive features that up to now has been manufactured by extraction from plants with long and not environmentally friendly procedures.

Black Crust Complex: Influence of Temperature and Period of Wetness on the Development of Fungi in Hevea brasiliensis.

The objective of this work was to evaluate the development of Davidiella sp. and its asexual form, Cladosporium sp., under different environmental conditions in the rubber tree (Hevea brasiliensis). Rubber tree leaves were inoculated with a spore suspension and kept in a humid chamber under different temperatures and wetness periods. The behavior of the fungi was evaluated using a scanning electron microscope (SEM) and an ultraviolet light microscope (UV). In the images obtained in SEM, four hours after inoculation of the fungus, it was possible to verify the germination and penetration of conidia at temperatures of 10 to 20 °C. The formation of conidiophores was verified from six hours after inoculation, indicating that it is in the reproductive period. In the sexual phase, in SEM, from four hours after inoculation, it was possible to verify the formation of small protuberances at temperatures between 10 and 20 °C. These black dots evolve into circular, protruding black spots, like the symptoms of black crust, with apparent spore formation on them. The data obtained from the UV analyses corroborate those from SEM, showing that the fungus has good development in its two phases between temperatures of 20 and 25 °C and that the period of wetness on the leaf can contribute to the initial development of the pathogen.

Antifungal Activity of Morinda citrifolia Methanolic Extract against Candida albicans: An In Vitro Study.

Aim: Natural medicine used as an alternative and/or complementary treatment to counteract diseases is of great importance in public health. Therefore, the purpose of the present study was to assess the in vitro antifungal activity of Morinda citrifolia methanolic extract of peel, pulp, and seed against Candida albicans. Materials and Methods: The present study was experimental in vitro and cross-sectional. Eight replicates were prepared in Sabouraud dextrose agar with five wells each, where 0.12% chlorhexidine, distilled water, and methanolic extract of seed, peel, and pulp of Morinda citrifolia fruit were placed at concentrations of 10,690, 8,270, and 6,430 mg/mL, respectively, to evaluate sensitivity according to Duraffourd's scale. In addition, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by dilution and agar seeding method. Statistical analysis was performed by analysis of variance (ANOVA) and Tukey's post hoc test, considering a significance level of P < 0.05. Results: The inhibition halos of Morinda citrifolia methanolic extract of seed, peel, and pulp against Candida albicans measured on average 15.94, 11.94, and 11.56 mm, respectively. The MIC of seed, peel, and pulp extract were 1366.25, 2067.5, and 1607.5 mg/mL respectively, whereas the MFC for seed, peel, and pulp extract were 2672.50, 2067.5, and 3215 mg/mL, respectively. Moreover, seed extract presented significantly higher antifungal activity than peel and pulp (P < 0.001). Conclusions: Morinda citrifolia methanolic extract of peel, pulp, and seed showed fungistatic and fungicidal effect against Candida albicans, being this very sensitive to seed extract with a MIC of 1366.25 mg/mL and a MFC of 2672.5 mg/mL, which allows recommending the development of effective pharmacological formulations for the control of candidiasis.

Candidiasis in Choloepus sp.-A Review of New Advances on the Disease.

Candidiasis is a significant fungal infection caused by various species of the genus Candida, posing health challenges to a wide range of animals, including Choloepus species (two-toed sloths). This review article aims to provide a comprehensive understanding of candidiasis in Choloepus sp., highlighting the etiology, epidemiology, pathogenesis, clinical manifestations, diagnosis, treatment, and prevention strategies. This article begins by examining the causative agents, primarily focusing on Candida albicans, which is the most commonly implicated species in candidiasis. The epidemiological aspects are discussed, emphasizing the prevalence of candidiasis in wild and captive Choloepus populations and identifying predisposing factors, such as immunosuppression, stress, poor nutrition, and environmental conditions. Pathogenesis is explored, detailing the mechanisms through which Candida species invade host tissues and evade immune responses. Clinical manifestations in Choloepus sp. are described, including oral thrush, cutaneous lesions, and gastrointestinal infections, and their impact on the health and behavior of affected individuals. Diagnostic methods, including culture techniques, histopathology, and molecular assays, are reviewed to highlight their roles in accurately identifying Candida infections. This article also covers treatment options, focusing on antifungal therapies and supportive care tailored to the unique physiology of Choloepus sp. Finally, prevention and management strategies are discussed, emphasizing the importance of maintaining optimal husbandry practices, regular health monitoring, and early intervention to reduce the incidence and impact of candidiasis in Choloepus populations. This review underscores the need for further research to enhance our understanding of candidiasis and improve health outcomes for these unique and vulnerable animals.

Comparative genomics and stable isotope analysis reveal the saprotrophic-pathogenic lifestyle of a neotropical fungus.

In terrestrial forested ecosystems, fungi may interact with trees in at least three distinct ways: (i) associated with roots as symbionts; (ii) as pathogens in roots, trunks, leaves, flowers, and fruits; or (iii) decomposing dead tree tissues on soil or even on dead tissues in living trees. Distinguishing the latter two nutrition modes is rather difficult in Hymenochaetaceae (Basidiomycota) species. Herein, we have used an integrative approach of comparative genomics, stable isotopes, host tree association, and bioclimatic data to investigate the lifestyle ecology of the scarcely known neotropical genus Phellinotus, focusing on the unique species Phellinotus piptadeniae. This species is strongly associated with living Piptadenia gonoacantha (Fabaceae) trees in the Atlantic Forest domain on a relatively high precipitation gradient. Phylogenomics resolved P. piptadeniae in a clade that also includes both plant pathogens and typical wood saprotrophs. Furthermore, both genome-predicted Carbohydrate-Active Enzymes (CAZy) and stable isotopes (δ13C and δ15N) revealed a rather flexible lifestyle for the species. Altogether, our findings suggest that P. piptadeniae has been undergoing a pathotrophic specialization in a particular tree species while maintaining all the metabolic repertoire of a wood saprothroph. IMPORTANCE: This is the first genomic description for Phellinotus piptadeniae. This basidiomycete is found across a broad range of climates and ecosystems in South America, including regions threatened by extensive agriculture. This fungus is also relevant considering its pathotrophic-saprotrophic association with Piptadenia goanocantha, which we began to understand with these new results that locate this species among biotrophic and necrotrophic fungi.

Purpureocillium lilacinum SBF054: Endophytic in Phaseolus vulgaris, Glycine max, and Helianthus annuus; Antagonistic to Rhizoctonia solani; and Virulent to Euschistus heros.

Microorganisms with multiple ecological functions can be a useful biotechnological resource in integrated pest- and disease-management programs. This work aimed to investigate the potential endophytic and virulent effects of a strain of Purpureocillium lilacinum on organic cultivation in Brazil. Specifically, the strain's ability to establish itself as an endophyte in common bean, soybean, and sunflower plants when inoculated via seed was evaluated. Furthermore, its antifungal activity against phytopathogens and its pathogenicity and virulence against insects of the order Lepidoptera, Coleoptera, and Hemiptera were evaluated. Furthermore, the strain was evaluated for its biochemical and physiological characteristics. For virulence bioassays, the experiments were conducted under a factorial scheme (2 × 3), with the following factors: (a) fungal inoculation and control without inoculum and (b) types of inocula (blastospores, aerial conidia, and metabolites). The treatments were sprayed on insect species at different stages of development. In summary, it was found that the SBF054 strain endophytically colonized the common bean, with partial recovery from the root tissues of soybean and sunflower plants, 30 days after inoculation; suppressed 86% of Rhizoctonia solani mycelial growth in an in vitro assay; and controlled eggs, nymphs, and Euschistus heros adults. These multifunctional abilities are mainly attributed to the strain's mechanisms of producing metabolites, such as organic acids, soluble nutrients, and hydrolytic enzymes.

Indigenous fungi with the ability to biodegrade hydrocarbons in diesel-contaminated soil are isolated and selected using a simple methodology.

Soil contamination by hydrocarbons is a problem that causes severe damage to the environment and public health. Technologies such as bioremediation using native microbial species represent a promising and environmentally friendly alternative for decontamination. This study aimed to isolate indigenous fungi species from the State of Rio de Janeiro, Brazil and evaluate their diesel degrading capacity in soils contaminated with crude oil. Seven filamentous fungi were isolated after enrichment cultivation from soils collected from contaminated sites and subjected to growth analysis on diesel nutrient media. Two fungal species were pre-selected and identified by morphological genus analysis and molecular techniques as Trichoderma asperellum and Penicillium pedernalense. The microdilution test showed that T. asperellum presented better fungal growth in high diesel concentrations than P. pedernalense. In addition, T. asperellum was able to degrade 41 and 54% of the total petroleum hydrocarbon (TPH) content present in soil artificially contaminated with diesel (10 g/kg of soil) in 7 and 14 days of incubation, respectively. In higher diesel concentration (1000 g of diesel/kg of soil) the TPH degradation reached 26%, 45%, and 48%, in 9, 16, and 30 d, respectively. The results demonstrated that the selected species was suitable for diesel degradation. We can also conclude that the isolation and selection process proposed in this work was successful and represents a simple alternative for obtaining native species with hydrocarbon degradation capacity, for use in the bioremediation process in the recovery of contaminated areas in an ecologically acceptable way.

Novel Pseudomonas Species Prevent the Growth of the Phytopathogenic Fungus Aspergillus flavus.

In response to the escalating demand for sustainable agricultural methodologies, the utilization of microbial volatile organic compounds (VOCs) as antagonists against phytopathogens has emerged as a viable eco-friendly alternative. Microbial volatiles exhibit rapid diffusion rates, facilitating prompt chemical interactions. Moreover, microorganisms possess the capacity to emit volatiles constitutively, as well as in response to biological interactions and environmental stimuli. In addition to volatile compounds, these bacteria demonstrate the ability to produce soluble metabolites with antifungal properties, such as APE Vf, pyoverdin, and fragin. In this study, we identified two Pseudomonas strains (BJa3 and MCal1) capable of inhibiting the in vitro mycelial growth of the phytopathogenic fungus Aspergillus flavus, which serves as the causal agent of diseases in sugarcane and maize. Utilizing GC/MS analysis, we detected 47 distinct VOCs which were produced by these bacterial strains. Notably, certain volatile compounds, including 1-heptoxydecane and tridecan-2-one, emerged as primary candidates for inhibiting fungal growth. These compounds belong to essential chemical classes previously documented for their antifungal activity, while others represent novel molecules. Furthermore, examination via confocal microscopy unveiled significant morphological alterations, particularly in the cell wall, of mycelia exposed to VOCs emitted by both Pseudomonas species. These findings underscore the potential of the identified BJa3 and MCal1 Pseudomonas strains as promising agents for fungal biocontrol in agricultural crops.

An Experience in Laboratory Diagnosis of Fungal Infections in COVID -19 Patients.

Introduction Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has cast a gloom spell on healthcare worldwide, infecting millions of people. Objective The aim of the present study is to determine the prevalence and review the contributing comorbidities and the precipitating factors leading to the emergence of the fungal infections in COVID-19-affected patients. To assess the utility of different laboratory techniques for confirmation of fungal infections. To assess the strengths and limitations of the diagnostic methods. Methods We have studied 252 clinical samples obtained from 121 COVID-positive patients. Results Among the 121 patients clinically diagnosed with fungal infections, 88 had diabetes and were given steroids for treatment ( p -value = 0.001). Ninety-five patients (78.5%) had a positive laboratory diagnosis (either culture positive, potassium hydroxide [KOH]-positive or positive histopathology report). Fungal culture was positive in 75 (61.9%) patients and histopathology report was positive in 62 (51.2%). Histopathology was positive in 7 (5.8%) patients in whom culture and KOH were negative. Conclusion Aggressive treatment methods, administration of immune suppressants, and antibiotics, with an intention to salvage, have made patients susceptible to the benign fungus, causing it to evade the host immunity, thus leading to invasive infections. Applying different laboratory modalities would not only aid in providing fast and valuable information but also help in understanding the pathology which would assist the clinician in selecting the correct treatment for the patient.

Hyperspectral imaging for the detection of plant pathogens in seeds: recent developments and challenges.

Food security, a critical concern amid global population growth, faces challenges in sustainable agricultural production due to significant yield losses caused by plant diseases, with a multitude of them caused by seedborne plant pathogen. With the expansion of the international seed market with global movement of this propagative plant material, and considering that about 90% of economically important crops grown from seeds, seed pathology emerged as an important discipline. Seed health testing is presently part of quality analysis and carried out by seed enterprises and governmental institutions looking forward to exclude a new pathogen in a country or site. The development of seedborne pathogens detection methods has been following the plant pathogen detection and diagnosis advances, from the use of cultivation on semi-selective media, to antibodies and DNA-based techniques. Hyperspectral imaging (HSI) associated with artificial intelligence can be considered the new frontier for seedborne pathogen detection with high accuracy in discriminating infected from healthy seeds. The development of the process consists of standardization of methods and protocols with the validation of spectral signatures for presence and incidence of contamined seeds. Concurrently, epidemiological studies correlating this information with disease outbreaks would help in determining the acceptable thresholds of seed contamination. Despite the high costs of equipment and the necessity for interdisciplinary collaboration, it is anticipated that health seed certifying programs and seed suppliers will benefit from the adoption of HSI techniques in the near future.

[Identification and sensitization to environmental fungal spores in Lima City, Peru]. / Identificación y sensibilización a hongos ambientales en Lima, Perú.

OBJECTIVE: To identify and registry the most important fungal spores trapped in our aerobiology station, as well as to report the prevalence of skin sensitization to these allergens. METHODS: The pollen counts were made according to standardized technique with a Burkard seven days spore trap, following the American Academy of Allergy, Asthma and Immunology (AAAAI) through National Allergy Bureau (NAB) recommendations. The trap was installed on the roof of Clinica SANNA, El GOLF, San Isidro, which is 20 m high, 12°5'54"S 77°3'6"W in the west-south of the Lima urban area. The sampling period was performed from September 2020 to October 2021. Skin prick tests were carried out according to the recommendations of the Spanish Society of Allergology and Clinical Immunology (SEAIC) in 200 patients (18 to 60 years old) with symptoms of rhinoconjunctivitis and/or asthma, who were evaluated in the Allergology Service of Clinica SANNA el Golf. Allergenic extracts were applied, dust mites (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis), cat and dog danders, cockroach (Periplaneta americana), grass 6 mix, weed mix, molds (Cladosporium herbarum, Alternaria alternata, Aspergillus fumigatus, Penicillium notatum, Nigrospora spp.), INMUNOTEK-Spain provided the extracts. We also tested other fungal allergens such as Fusarium spp, Stemphylium spp, Curvularia spp, a mixture of Helmintosporum/Dreschlera spp. from the DIATER-Argentina laboratory. RESULTS: We identified spores of Alternaria alternata, Cladosporium spp., Nigrospora spp., Stemphylium spp., Fusarium spp., Curvularia spp., Dreschlera/Helmintosporum spp. The patients showed sensitization to Cladosporium herbarum (14%), Fusarium spp. (13,5%), Nigrospora spp. (8%), Alternaria Alternata (7%), Stemphylium (6%), Dreschlera/Helmintosporium spp. (5,5%), Curvularia spp. (3%), Aspergillus fumigatus (2,5%). CONCLUSIONS: The inhabitants of the south-western area of the urban city of Lima are exposed to different fungal spores with allergenic potential, with a higher concentration being identified during the summer/autumn months. Cutaneous sensitization is demonstrated in variable percentages to the fungal spores identified in this aerobiological sampling. The results of this study should be expanded and compared with data in the forthcoming years, identify seasonal and annual fluctuations and extend the traps to other locations in Lima.

OBJETIVO: Identificar y registrar las esporas de hongos más importantes captadas en nuestra estación de aerobiología, además reportar la prevalencia de sensibilización cutánea a estos alérgenos. MÉTODOS: La identificación y los conteos de esporas de hongos se realizaron según la técnica estandarizada con un equipo colector Burkard Spore Trap For Seven Days, siguiendo las recomendaciones de la National Allergy Bureau (NAB), de la American Academy Allergy Asthma and Immunology (AAAAI). El equipo se instaló a 20 m de altura desde el nivel del suelo, en la azotea de la Clínica SANNA El Golf, distrito de San Isidro, (12°5'54"S 77°3'6"O), en la zona sur-oeste del área urbana de Lima. El periodo de captación se llevó a cabo entre septiembre de 2020 y octubre de 2021. Se realizaron estudios de pruebas cutáneas (skin prick-test), según recomendaciones de la Sociedad Española de Alergología e Inmunología Clínica (SEAIC), en 200 pacientes (entre 18 y 60 años), con sintomatología de rinoconjuntivitis y/o asma. Fueron evaluados en el servicio de Alergología de la Clínica SANNA El Golf. Se aplicaron extractos alergénicos de ácaros del polvo (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis), epitelios de gato y perro, Periplaneta americana, mezclas de seis gramíneas, mezclas de malezas, hongos ambientales (Cladosporium herbarum, Alternaria alternata, Aspergillus fumigatus, Penicillium notatum, Nigrospora spp.), extractos del laboratorio INMUNOTEK-España. Además, testeamos otros alérgenos fúngicos de Fusarium spp, Stemphylium spp, Curvularia spp, una mezcla de Helmintosporum/Dreschlera spp. del laboratorio DIATER-Argentina. RESULTADOS: Identificamos esporas de Alternaria alternata, Cladosporium spp., Nigrospora spp., Stemphylium spp., Fusarium spp., Curvularia spp., Dreschlera/Helmintosporum spp. Los pacientes mostraron sensibilización a Cladosporium herbarum (14%), Fusarium spp. (13,5%), Nigrospora spp. (8%), Alternaria Alternata (7%), Stemphylium (6%), Dreschlera/Helmintosporium spp. (5,5%), Curvularia spp. (3%) y Aspergillus fumigatus (2,5%). CONCLUSIONES: Los habitantes de la zona sur-oeste de la ciudad urbana de Lima están expuestos a distintas esporas de hongos con potencial alergénico, identificándose mayor concentración durante los meses de verano y otoño. Se demuestra sensibilización cutánea en porcentajes variables a las esporas fúngicas identificadas en este muestreo aerobiológico. Los resultados de este estudio deberían ampliarse y ser comparados con data en los años siguientes, identificar fluctuaciones estacionales y anuales y extender los captadores a otras locaciones en Lima.