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In northern Tamaulipas, México, the contamination of corn by toxigenic fungi reduces grain production and quality. Corn contaminated by mycotoxins puts humans and livestock at risk. Continuous monitoring of the sanitary quality of grain at harvest and in storage will define preventive and corrective strategies for contamination by mycotoxigenic fungi. In this work, we identified toxigenic fungi associated with corn grown and stored in northern Tamaulipas, identified and quantified aflatoxins and their relationships with the physicochemical characteristics of the grain, and identified the main genes responsible for aflatoxin production in A. flavus. Fungal incidence was evaluated in vitro, aflatoxin production was evaluated via HPLC, and physicochemical traits were evaluated via spectrophotometry. Three genera were identified: Fusarium, Aspergillus, and Penicillium; the latter had the highest incidence in both 2011 and 2012. The incidence was higher in 2012 (82.3%) than in 2011 (4.5%), and storage did not affect the incidence. Associations among fungal incidences and physicochemical traits were significant and intermediate in both years. AFB1 production was negatively associated with hectoliter weight, and total fungal incidence was positively related to the incidence of Penicillium, Fusarium, and Aspergillus and negatively related to the flotation index. AFB1 was detected in 13.18% of the samples, with values ranging from 3.4881.33 ppb upon receipt and from 4.3245.92 ppb after storage. Two samples exceeded the allowed limits for Mexico (20 ppb). The aflD and aflQ genes were detected in 52.1 and 56.3%, respectively, of the A. flavus isolates. [ABSTRACT FROM AUTHOR]

Cereals are consumed globally because of their nutritional values and potential to reduce malnutrition. Despite their usefulness, cereals are prone to aflatoxin contamination. Aflatoxins are highly toxic and carcinogenic secondary metabolic products that contaminate agricultural products consumed by humans. Studies have shown that aflatoxin is found in cereals at a high level. Human exposure to aflatoxin through food and feed results in a wide range of health issues, including a weakened immune system and cancer. Worst, it can cause death depending on the level and extent of exposure. Several climate-induced factors, such as drought, can trigger aflatoxin production worldwide, especially in Africa, where the environment is conducive. Several precautions have been taken to mitigate human exposure to aflatoxin, including strict regulations, pre- and post-harvest contamination prevention, detoxification, and decontamination. In addition, good farm management and practices, and awareness creation and education can help to reduce aflatoxin contamination and exposure, if not eradicate it. The current review detailed the up-to-date information on aflatoxin occurrence, health implications, and control measures for aflatoxin in cereals to ensure food safety and human well-being. This review also illuminated the potential hazards of human exposure to aflatoxin beyond the permissible level, particularly in children. [ABSTRACT FROM AUTHOR]

Purpose: To identify pathogenic microorganisms and microbiological risk factors causing high morbidity and mortality in immunocompromised patients requiring invasive mechanical ventilation due to pneumonia. Methods: A retrospective single-center study was performed at the intensive care unit (ICU) of the Department of Internal Medicine at Heidelberg University Hospital (Germany) including 246 consecutive patients with hematological malignancies requiring invasive mechanical ventilation due to pneumonia from 08/2004 to 07/2016. Microbiological and radiological data were collected and statistically analyzed for risk factors for ICU and 1-year mortality. Results: ICU and 1-year mortality were 63.0% (155/246) and 81.0% (196/242), respectively. Pneumonia causing pathogens were identified in 143 (58.1%) patients, multimicrobial infections were present in 51 (20.7%) patients. Fungal, bacterial and viral pathogens were detected in 89 (36.2%), 55 (22.4%) and 41 (16.7%) patients, respectively. Human herpesviruses were concomitantly reactivated in 85 (34.6%) patients. As significant microbiological risk factors for ICU mortality probable invasive Aspergillus disease with positive serum-Galactomannan (odds ratio 3.1 (1.2-8.0), p = 0.021,) and pulmonary Cytomegalovirus reactivation at intubation (odds ratio 5.3 (1.1–26.8), p = 0.043,) were identified. 1-year mortality was not significantly associated with type of infection. Of interest, 19 patients had infections with various respiratory viruses and Aspergillus spp. superinfections and experienced high ICU and 1-year mortality of 78.9% (15/19) and 89.5% (17/19), respectively. Conclusions: Patients with hematological malignancies requiring invasive mechanical ventilation due to pneumonia showed high ICU and 1-year mortality. Pulmonary Aspergillosis and pulmonary reactivation of Cytomegalovirus at intubation were significantly associated with negative outcome. [ABSTRACT FROM AUTHOR]

Abstract Background Aspergillus species cause a variety of serious clinical conditions with increasing trend in antifungal resistance. The present study aimed at evaluating hospital epidemiology and antifungal susceptibility of all isolates recorded in our clinical database since its implementation. Methods Data on date of isolation, biological samples, patients’ age and sex, clinical settings, and antifungal susceptibility tests for all Aspergillus spp. isolated from 2015 to 2022 were extracted from the clinical database. Score test for trend of odds, non-parametric Mann Kendall trend test and logistic regression analysis were used to analyze prevalence, incidence, and seasonality of Aspergillus spp. isolates. Results A total of 1126 Aspergillus spp. isolates were evaluated. A. fumigatus was the most prevalent (44.1%) followed by A. niger (22.3%), A. flavus (17.7%) and A. terreus (10.6%). A. niger prevalence increased over time in intensive care units (p-trend = 0.0051). Overall, 16 (1.5%) were not susceptible to one azole compound, and 108 (10.9%) to amphotericin B, with A. niger showing the highest percentage (21.9%). The risk of detecting A. fumigatus was higher in June, (OR = 2.14, 95% CI [1.16; 3.98] p = 0.016) and reduced during September (OR = 0.48, 95% CI [0.27; 0.87] p = 0.015) and October as compared to January (OR = 0.39, 95% CI [0.21; 0.70] p = 0.002. A. niger showed a reduced risk of isolation from all clinical samples in the month of June as compared to January (OR = 0.34, 95% CI [0.14; 0.79] p = 0.012). Seasonal trend for A. flavus showed a higher risk of detection in September (OR = 2.7, 95% CI [1.18; 6.18] p = 0.019), October (OR = 2.32, 95% CI [1.01; 5.35] p = 0.048) and November (OR = 2.42, 95% CI [1.01; 5.79] p = 0.047) as compared to January. Conclusions This is the first study to analyze, at once, data regarding prevalence, time trends, seasonality, species distribution and antifungal susceptibility profiles of all Aspergillus spp. isolates over a 8-year period in a tertiary care center. Surprisingly no increase in azole resistance was observed over time.

Cereals and legumes are the major food sources for people in a developing country. Four grains and legumes (rice, maize wheat and groundnut) stored for 2 - 4 months in different packaging materials. These samples were randomly selected from three different markets. They were assessed for the presence of mycotoxin producing moulds and for the production of mycotoxins. Standard microbiological and molecular methods were used in the isolation and identification of moulds. A multimycotoxin method based on Liquid Chromatography tandem mass spectrometry was used to analyze both the qualitative and quantitative occurrence of mycotoxin fungal metabolites. Proximate composition was determined using the method of Association of official Analytical chemist. The moulds isolated and identified culturally were Aspergillus niger, Aspergillus flavus, Aspergillus spp., Aspergillus tamarii Penicillium chrysogenum, Fusarium spp., Rhizopus stolonifer, Rhizopus nigricans and Mucor spp. The percentage occurrence of non-culturally 18S rRNA gene sequence dominant mould species identified were Aspergillus flavus (46%) followed by Aspergillus tamarii (23%), Aspergillus niger (18%), and Penicillium chrysogenum (9%) while the least was (4%) Aspergillus brunneoviolaceus. The Phylogenic tree was constructed by using the geneious software version 4.0. Aflatoxin, ochratoxins, fumonisin, deoxynevalenol and zearalenone were the different mycotoxins detected in stored grains and legumes. Ochratoxin A had the highest concentration of 371.8 ± 7.878 while Deoxynevalenol had the least concentration of 320 ± 4.617. Different values for Moisture Content, Crude Protein, Crude Fibre, Ash, Carbohydrate and Energy Value were determined. Groundnut 558.74 ± 279.37 had the highest energy value while Wheat 315.08 ± 157.54 had the least energy value. Grains and Legumes are essential for good health. There is a strong need to devise good storage condition for stored grains and legumes to avoid mycotoxigenic moulds contamination. [ABSTRACT FROM AUTHOR]

Background: Aspergillus species cause a variety of serious clinical conditions with increasing trend in antifungal resistance. The present study aimed at evaluating hospital epidemiology and antifungal susceptibility of all isolates recorded in our clinical database since its implementation. Methods: Data on date of isolation, biological samples, patients' age and sex, clinical settings, and antifungal susceptibility tests for all Aspergillus spp. isolated from 2015 to 2022 were extracted from the clinical database. Score test for trend of odds, non-parametric Mann Kendall trend test and logistic regression analysis were used to analyze prevalence, incidence, and seasonality of Aspergillus spp. isolates. Results: A total of 1126 Aspergillus spp. isolates were evaluated. A. fumigatus was the most prevalent (44.1%) followed by A. niger (22.3%), A. flavus (17.7%) and A. terreus (10.6%). A. niger prevalence increased over time in intensive care units (p-trend = 0.0051). Overall, 16 (1.5%) were not susceptible to one azole compound, and 108 (10.9%) to amphotericin B, with A. niger showing the highest percentage (21.9%). The risk of detecting A. fumigatus was higher in June, (OR = 2.14, 95% CI [1.16; 3.98] p = 0.016) and reduced during September (OR = 0.48, 95% CI [0.27; 0.87] p = 0.015) and October as compared to January (OR = 0.39, 95% CI [0.21; 0.70] p = 0.002. A. niger showed a reduced risk of isolation from all clinical samples in the month of June as compared to January (OR = 0.34, 95% CI [0.14; 0.79] p = 0.012). Seasonal trend for A. flavus showed a higher risk of detection in September (OR = 2.7, 95% CI [1.18; 6.18] p = 0.019), October (OR = 2.32, 95% CI [1.01; 5.35] p = 0.048) and November (OR = 2.42, 95% CI [1.01; 5.79] p = 0.047) as compared to January. Conclusions: This is the first study to analyze, at once, data regarding prevalence, time trends, seasonality, species distribution and antifungal susceptibility profiles of all Aspergillus spp. isolates over a 8-year period in a tertiary care center. Surprisingly no increase in azole resistance was observed over time. [ABSTRACT FROM AUTHOR]

This work aimed to investigate the prevalence of pathogenic fungi and evaluate the antifungal activity of Trichoderma orientale FMR12486 crude extract against pathogenic fungi isolated from patients attending the National Center for Thoracic and Respiratory Diseases (having a history of tuberculosis) and consultant of Dermatology of Baghdad hospital, Iraq. A total of 80 clinical specimens were collected: 20 skin scrapings specimens and 60 sputum specimens. The results of direct examination by KOH 10% and culture showed that 11 (55%) cases from 20 skin specimens were positive for fungal infections, while in the sputum specimens, 28 (47%) cases from 60 were positive. Candida albicans represented the most common fungal infection isolated from sputum specimens which represented 18 cases at a rate of 64.2%, followed by Candida tropicalis 5 (17.9%), Aspergillus fumigates 2 (7.1%), Aspergillus flavus 1 (3.6%), Candida glabrata 1 (3.6%) and Candida krusei 1 (3.6%). While in the skin scrapes, Candida parapsilosis was the common which represented 3 cases at a rate of 27.3%, followed by Trichophyton mentagrophyte 2 (18.2%), Microsporium canis 2 (18.2%), C. albicans 2 (18.2%), Trichophyton rubrum 1 (9.1%), C. tropicalis 1 (9.1%). T. orientale isolates were collected from the gardens soil of the University of Baghdad and were then identified depending on the morphological feature of the colony and microscopic characteristics. To confirm identification, PCR technique was used in which DNA of T. orientale was extracted at a concentration of 54 - 294 μg/2 g wet mycelium with a purity of 1.6 - 1.8. DNA samples were amplified with primers ITS-1and ITS-4, And the results confirmed that the isolate was T. orientale strain FMR12486. This isolate was processed with ethyl acetate using a standard extraction method and the final amount of crude extract being 6 grams. Different concentrations of crude extract 0.5, 1, 2 and 4 mg/ml were prepared and used against fungal pathogens isolated using the agar-well diffusion method. The results showed high antifungal activity against pathogenic isolates, significantly recording maximum inhibition zones of 20 and 22 mm against C. parapsilosis at concentrations of 0.5 and 1 mg/ml, respectively. It was followed by T. mentagrophyte and C. glabrata. 16, 20, and 16, 20 mm that were recorded at concentrations of 2 and 4mg/ml, a significant maximum inhibition zone was recorded 26 mm against C. glabrata, followed by C. parapsilosis, M. canis and T. mentagrophyte (24, 26; 16, 24 and 20, 24mm respectively). Just Trichophyton rubrum fungus in this study showed resistance to all concentrations of the crude extract where the inhibition zone was zero. The results of the current study showed that T. orientale strain FMR12486 crude extract can be used to make an effective drug to treat human fungal infections. [ABSTRACT FROM AUTHOR]

The study of pathogenicity and virulence of fungal strains, in vivo in the preclinical phase, is carried out through the use of animal models belonging to various classes of mammals (rodents, leproids, etc.). Although animals are functionally more similar to humans, these studies have some limitations in terms of ethics (animal suffering), user-friendliness, cost-effectiveness, timing (physiological response time) and logistics (need for adequately equipped laboratories). A good in vivo model must possess some optimal characteristics to be used, such as rapid growth, small size and short life cycle. For this reason, insects, such as Galleria mellonella (Lepidoptera), Drosophila melanogaster (Diptera) and Bombyx mori (Lepidoptera), have been widely used as alternative non-mammalian models. Due to their simplicity of use and low cost, the larvae of G. mellonella represent an optimal model above all to evaluate the virulence of fungal pathogens and the use of antifungal treatments (either single or in combination with biologically active compounds). A further advantage is also represented by their simple neuronal system limiting the suffering of the animal itself, their ability to survive at near-body ambient temperatures as well as the expression of proteins able to recognise combined pathogens following the three R principles (replacement, refinement and reduction). This review aims to assess the validity as well as the advantages and disadvantages of replacing mammalian classes with G. mellonella as an in vivo study model for preclinical experimentation. [ABSTRACT FROM AUTHOR]

Pulmonary aspergillosis constitutes an increasingly prevalent and potentially fatal complex of mycotic diseases, caused by different species of Aspergillus. The broad spectrum of pathological manifestations associated with pulmonary aspergillosis necessitates a differentiation of commensalism from saprophytic colonization, hypersensitivity reactions, and true invasive infections, which highlights the importance of histopathology as a gold standard in a diagnostic setting. For the past decades, changes in terminology and contradicting contributions from different diagnostic disciplines have made the classification of pulmonary aspergillosis rather confusing. This review offers a categorization of aspergillosis lesions based on what can be histopathologically identified and distinguished, differentiating between acute invasive infection and forms of subacute, chronic, and allergic diseases and coinfections, and summarizes important manifestations of lesions associated with the different forms of pulmonary aspergillosis. [ABSTRACT FROM AUTHOR]

Introduction: Secondary bacterial and fungal infections are a significant concern in COVID-19 patients, particularly those critically ill and requiring intensive care. This retrospective study investigated the prevalence and spectrum of secondary infections among COVID-19 patients admitted to the intensive care unit (ICU) at a tertiary care hospital in Navi Mumbai. Additionally, we explored the association between secondary infections and patient comorbidities. Methods: We performed a single-center, retrospective cohort study of 3234 COVID-19 patients admitted to a tertiary care hospital in Navi Mumbai, India, between August 2020 and August 2021. Microbiological data from various clinical specimens, including blood, sputum, bronchoalveolar lavage (BAL) fluid, urine, and tissue cultures, were retrospectively analyzed. Patient demographics and comorbidities were extracted from medical records. We employed descriptive statistics and Pearson's Chi-square test for data analysis to identify associations between secondary infections and patient characteristics. Results: Among the 3234 COVID-19 patients, 195 (6.02%) presented with clinical features suggestive of secondary infections. Microbiological analysis confirmed secondary infections in 98 patients (3.03%), with a culture positivity rate of 50.3%. Among bacterial isolates, Klebsiella pneumoniae was the most prevalent (43.28%), followed by Acinetobacter baumannii (25.37%). Aspergillus spp. emerged as the dominant fungal pathogen. Notably, Escherichia coli isolation was significantly associated with various specimen types (P < 0.001). However, no significant correlation was found between secondary infection rates and patient comorbidities. Conclusion: Gram-negative bacteria, specifically K. pneumoniae and A. baumannii, were the primary pathogens responsible for secondary infections in our cohort of critically ill COVID-19 patients admitted to the ICU. These findings underscore the importance of ongoing surveillance and monitoring of secondary infection trends, including fungal pathogens, to inform and optimize management strategies in this high-risk population. [ABSTRACT FROM AUTHOR]

Contamination of cereals with fungi Aspergillus spp. and mycotoxins aflatoxins causes losses in the agrofood chain and cancers in animals and humans. This article assesses the occurrence of total aflatoxin contamination in common wheat in Romania during the 2015 - 2016 period under the influence of environmental factors and extreme weather events. Wheat samples (N = 272) taken from localities characterized by deoxynivalenol contamination in 2012 - 2014, were analyzed for aflatoxins using the enzyme-linked immunosorbent assay (ELISA) method and gained resuts were statistically evaluated by SPSS. Total aflatoxin contamination in common wheat was very low (max. 1.10 µg/kg) in June - August of the 2015 extremely dry year, but high values were recorded in Covasna, Harghita, and Vâlcea counties (4.00 µg/kg; 5.42 µg/kg; 4.58 µg/kg; acidic soils) in May - June 2016, an extremely rainy year. Extremely hot and dry weather conditions in July - August 2015 reduced the effects of environmental factors to non-significant correlations with total aflatoxins. The effects of the extreme drought were also manifested in 2016, but the temperature and precipitation in July - August were significantly correlated with aflatoxin contamination. The results show the occurrence of total aflatoxin contamination in common wheat grown in regions characterized by Fusarium spp. and deoxynivalenol contamination in the year following a dry year. The increased frequency and intensity of extreme weather events in the last two decades and the predicted climate changes require the analysis of aflatoxins in wheat and other cereals grown in agricultural regions with low aridity. [ABSTRACT FROM AUTHOR]

Among the co-infectious agents in COVID-19 patients, Aspergillus species cause invasive pulmonary aspergillosis (IPA). IPA is difficult to diagnose and is associated with high morbidity and mortality. This study is aimed at identifying Aspergillus spp. from sputum and tracheal aspirate (TA) samples of COVID-19 patients and at determining their antifungal susceptibility profiles. A total of 50 patients with COVID-19 hospitalized in their intensive care units (ICU) were included in the study. Identification of Aspergillus isolates was performed by phenotypic and molecular methods. ECMM/ISHAM consensus criteria were used for IPA case definitions. The antifungal susceptibility profiles of isolates were determined by the microdilution method. Aspergillus spp. was detected in 35 (70%) of the clinical samples. Among the Aspergillus spp., 20 (57.1%) A. fumigatus, six (17.1%) A. flavus, four (11.4%) A. niger, three (8.6%) A. terreus, and two (5.7%) A. welwitschiae were identified. In general, Aspergillus isolates were susceptible to the tested antifungal agents. In the study, nine patients were diagnosed with possible IPA, 11 patients were diagnosed with probable IPA, and 15 patients were diagnosed with Aspergillus colonization according to the used algorithms. Serum galactomannan antigen positivity was found in 11 of the patients diagnosed with IPA. Our results provide data on the incidence of IPA, identification of Aspergillus spp., and its susceptibility profiles in critically ill COVID-19 patients. Prospective studies are needed for a faster diagnosis or antifungal prophylaxis to manage the poor prognosis of IPA and reduce the risk of mortality. [ABSTRACT FROM AUTHOR]

Introduction. Allergy to microscopic fungi (micromycetes), which can colonize the respiratory tract, is associated with an eosinophilic endotype of inflammation and severe disease course. Among the hallmarks of fungal allergy, experts identify fixed airways obstruction, bronchiectasis, and fibrosis. Currently, there has been proposed the term “allergic fungal airways disease” (AFAD), which combines various clinical phenotypes, which pathogenesis is based on IgE-mediated allergic reaction against fungal spores colonizing the respiratory tract. According to this approach, the AFAD group includes severe asthma with fungal sensitization and allergic bronchopulmonary aspergillosis (ABPA). A promising direction in AFAD treatment implies the use of genetically engineered biological drugs against T-2 inflammation. The aim of the study was to evaluate the prevalence ofAspergillusspp. sensitization and allergic bronchopulmonaryAspergillosis in patients with asthma. Materials and methods. A retrospective single-center registry study was conducted in the I.I. Mechnikov North-Western State Medical University, Ministry of Health of Russian Federation. The study included 523 patients with asthma of varying severity. The level of total and specific blood serum IgE against aeroallergens, includingAspergillusspp., was analyzed by enzyme immunoassay. The diagnosis of asthma was established in accordance with the recommendations of the GINA working group and the Clinical recommendations of the Ministry of Health of the Russian Federation, fungal sensitization and ABPA — in accordance with the recommendations of the ISHAM working group.Results. During the examination of patients with asthma, sensitization toAspergillusspp. recorded in 16.6% cases, severe asthma with sensitization toAspergillusspp. — in 7.4%, ABPA — in 6.1%. In patients with allergic asthma, the prevalence of sensitization toAspergillusspp. was 22%, severe asthma with sensitization toAspergillusspp. — 10.1%, ABLA — 8.1%. Among patients with ABPA, females (64%) aged 40 years and older prevailed. The major clinical manifestations of ABPA were presented as cough (96%) and shortness of breath (76%), CT scan radiological changes were in a form of bronchiectasis (72%). Conclusion. A high prevalence of sensitization toAspergillusspp. in patients with asthma was found. It is necessary to includeAspergillusspp. into the list of tested allergens while examining patients with chronic lung diseases for timely detection of AFAD and prescription of proper anti-inflammatory or antimycotic therapy.

IntroductionIt is of upmost importance to contribute to fill the knowledge gap concerning the characterization of the occupational exposure to microbial agents in the waste sorting setting (automated and manual sorting).MethodsThis study intends to apply a comprehensive field sampling and laboratory protocol (culture based-methods and molecular tools), assess fungal azole resistance, as well as to elucidate on potential exposure related health effects (cytotoxicity analyses). Skin-biota samples (eSwabs) were performed on workers and controls to identify other exposure routes.ResultsIn personal filter samples the guidelines in one automated industry surpassed the guidelines for fungi. Seasonal influence on viable microbial contamination including fungi with reduced susceptibility to the tested azoles was observed, besides the observed reduced susceptibility of pathogens of critical priority (Mucorales and Fusarium sp.). Aspergillus sections with potential toxigenic effect and with clinical relevance were also detected in all the sampling methods.DiscussionThe results regarding skin-biota in both controls´ and workers´ hands claim attention for the possible exposure due to hand to face/mouth contact. This study allowed concluding that working in automated and manual waste sorting plants imply high exposure to microbial agents.

The genus Aspergillus harbors human infection-causing pathogens and is involved in the complex one-health challenge of antifungal resistance. Here, a 6-year retrospective study was conducted with Aspergillus spp. isolated from patients with invasive, chronic, and clinically suspected aspergillosis in a tertiary teaching hospital. A total of 64 Aspergillus spp. clinical isolates were investigated regarding molecular identification, biofilm, virulence in Galleria mellonella, antifungal susceptibility, and resistance to amphotericin B and azoles. Aspergillus section Fumigati (A. fumigatus sensu stricto, 62.5%) and section Flavi (A. flavus, 20.3%; A. parasiticus, 14%; and A. tamarii, 3.1%) have been identified. Aspergillus section Flavi clinical isolates were more virulent than section Fumigati clinical isolates. Furthermore, scant evidence supports a link between biofilm formation and virulence. The susceptibility of the Aspergillus spp. clinical isolates to itraconazole, posaconazole, voriconazole, and amphotericin B was evaluated. Most Aspergillus spp. clinical isolates (67.2%) had an AMB MIC value equal to or above 2 µg/mL, warning of a higher probability of therapeutic failure in the region under study. In general, the triazoles presented MIC values above the epidemiological cutoff value. The high triazole MIC values of A. fumigatus s.s. clinical isolates were investigated by sequencing the promoter region and cyp51A locus. The Cyp51A amino acid substitutions F46Y, M172V, N248T, N248K, D255E, and E427K were globally detected in 47.5% of A. fumigatus s.s. clinical isolates, and most of them are associated with high triazole MICs. Even so, the findings support voriconazole or itraconazole as the first therapeutic choice for treating Aspergillus infections. This study emphasizes the significance of continued surveillance of Aspergillus spp. infections to help overcome the gap in knowledge of the global fungal burden of infections and antifungal resistance, supporting public health initiatives. [ABSTRACT FROM AUTHOR]

The province of Isabela is the top maize producer in the Philippines. The intensive cultivation and the tropical climate in the region may favour fungal growth and aflatoxin contamination in maize grains. Thus, the study aimed to determine the occurrence of aflatoxin and mycotoxigenic Aspergillus species in maize varieties in this region. Samples were obtained in six municipalities from hybrid (Bt) maize (n = 101) and open‐pollinated varieties (n = 6) during the dry season (March and April) of 2019. Aflatoxin levels were quantified through enzyme‐linked immunosorbent assay, and Aspergillus species were identified through cultural and molecular methods. Aflatoxin was detected in 50.5% of maize samples; 49.5% of samples were less than the limit of detection (3 μg/kg), 16.8% with 3–20 μg/kg, 10.9% with 21–50 μg/kg and 22.8% above 50 μg/kg. Samples within the acceptable level were 66.3% for food (<20 μg/kg) and 77.2% for animal feed (<50 μg/kg), while 22.8% of samples were above the acceptable level of the Philippine National Standard for raw maize grains. More than 90% of Aspergillus species detected were A. flavus. Other species identified were A. tamarii and A. terreus. Despite the dry production season in the province with low relative humidity during harvesting, inadequate post‐harvest practices and the presence of A. flavus elevated the level of aflatoxin in sample grains. Additional work involving multi‐year surveys is needed to confirm the results and conclusions of this study. [ABSTRACT FROM AUTHOR]

An antifungal agent can either kill or inhibit the growth of fungi by interfering with the formation of fungal cell membrane, weakening it and hindering cell division. Antifungal agents of amphotericin B, ketoconazole, fluconazole and voriconazole of Thermo Fisher Scientific limited were used for this study. Cultural analysis of stored grains and legumes (rice, maize, wheat, groundnut and beans) from Imo State was done using streak plate method. Sabouraud dextrose agar was used for the culture while Mueller Hinton agar was used for Antifungal sensitivity test. Moulds were further identified using 18s rRNA gene sequencing method. The antifungal sensitivity profile of isolated and identified moulds was evaluated using the clinical laboratory and standard institute approved methods for testing of moulds, the disk diffusion and tetrazolium chloride test. The results showed that Aspergillus flavus, Aspergillus tamarii, Aspergillus niger, Aspergillus brunneoviolaceus and Penicillium chrysogenum were the moulds isolated and identified using both cultural and 18S rRNA sequence. The fungal isolates were susceptible to ketoconazole and voriconazole. Amphotericin B was both resistant and susceptible to different moulds. The fungal isolates were resistant to fluconazole. Inhibition effects were more with the antifungal disc than with tetrazolium salts. All the isolates were resistant to tetrazolium chloride and gave no zone of inhibition. Combination of antifungal agent and tetrazolium chloride showed sensitivity only to ketoconazole. Antifungal disc alone gave a better zone of inhibition than the combination of antifungal agents with tetrazolium salts. This study showed that ketoconazole has greater inhibitory potential than other antifungal agents. Ketoconazole remains the best drug of choice among the studied antifungal agents for fungal infections. Therefore antifungal drugs can be used against moulds of economic importance in the country. [ABSTRACT FROM AUTHOR]

The study of pathogenicity and virulence of fungal strains, in vivo in the preclinical phase, is carried out through the use of animal models belonging to various classes of mammals (rodents, leproids, etc.). Although animals are functionally more similar to humans, these studies have some limitations in terms of ethics (animal suffering), user-friendliness, cost-effectiveness, timing (physiological response time) and logistics (need for adequately equipped laboratories). A good in vivo model must possess some optimal characteristics to be used, such as rapid growth, small size and short life cycle. For this reason, insects, such as Galleria mellonella (Lepidoptera), Drosophila melanogaster (Diptera) and Bombyx mori (Lepidoptera), have been widely used as alternative non-mammalian models. Due to their simplicity of use and low cost, the larvae of G. mellonella represent an optimal model above all to evaluate the virulence of fungal pathogens and the use of antifungal treatments (either single or in combination with biologically active compounds). A further advantage is also represented by their simple neuronal system limiting the suffering of the animal itself, their ability to survive at near-body ambient temperatures as well as the expression of proteins able to recognise combined pathogens following the three R principles (replacement, refinement and reduction). This review aims to assess the validity as well as the advantages and disadvantages of replacing mammalian classes with G. mellonella as an in vivo study model for preclinical experimentation.

The microbiological characteristics of the grapes are made up of a wide variety of microorganisms, including filamentous fungi. Their presence in grapes is traditionally associated with deterioration in quality. The health of the grapes is very important for obtaining quality wine. The objective of this study was to investigate the diversity of mycobiota on the surface and inside of different grapevine varieties at harvest time in the temperate climate of Slovakia and to identify potentially pathogenic isolates of Aspergillus and Penicillium producing selected mycotoxins. During the 2021 grape harvest, grapes were collected from the Small Carpathians wine region. Eleven grape samples were analyzed by the plating method and plating method with surface disinfection. Emphasis was placed on Aspergillus and Penicillium species because of their importance in mycotoxin production. Of the 605 fungal strains detected, 11 genera were identified in the exogenous mycobiota. The most common and abundant genera were Alternaria and Botrytis. In the genus Aspergillus, A. section Nigri is the most abundant, while in the genus Penicillium, P. raistrickii reached the highest frequency and abundance. Of the 379 strains detected and identified from the endogenous mycobiota, the most common genera were again Alternaria and Botrytis and the most abundant genus was Botrytis. Penicillium species were detected in 17% of all fungi found, with P. raistrickii dominating. The A. section Nigri reached only 4% of the relative density of all isolates. Potentially toxigenic Aspergillus and Penicillium species were tested for toxinogenity by thin layer chromatography. The most important mycotoxin-producing species found were A. section Nigri but without ochratoxin A production.

A systematic study was carried out on 638 wheat and paddy grains (including fresh and stored samples) collected in 2021 from Shanghai, China, to identify the major mycobiota and their toxigenic abilities. A total of 349 fungi, namely, 252 Fusarium, 53 Aspergillus, and 44 Alternaria, were characterized by morphological and molecular identification. Fusarium and Aspergillus were more frequently isolated in paddy with Fusarium sambucinum species complex and Aspergillus section flavi as the predominant species, respectively. The genus Alternaria was the most frequently isolated fungal species in wheat. The toxin-producing potentials of the identified fungi were further evaluated in vitro. Deoxynevalenol (DON) was produced by 34.5% of Fusarium isolates and zearalenone (ZEN) was produced by 47.6% of them, and one isolate also processed the abilities for fumonisin B1 (FB1), B2 (FB2), and B3 (FB3) productions. Aflatoxin B1 (AFB1), B2 (AFB2), and G1 (AFG1) were only generated by Aspergillus section flavi, with the production rate of 65.5%, 27.6%, and 13.8%, respectively. Alternariol (AOH) was the most prevalent Alternaria toxin, which could be produced by 95.5% of the isolates, followed by alternariol monomethyl ether (AME) (72.7%), altenuene (ALT) (52.3%), tenuazonic acid (TeA) (45.5%), tentoxin (TEN) (29.5%), and altenusin (ALS) (4.5%). A combinational analysis of mycobiota and toxigenic ability allowed us to provide comprehensive information about the production mechanisms of mycotoxins in wheat and paddy in a specific geographic area, and will be helpful for developing efficient prevention and control programs. [ABSTRACT FROM AUTHOR]

Substantial yield losses have been reported during the storage of Allium cepa L., and effective methods to minimize postharvest losses are currently lacking. Therefore the objectives of this study were to investigate the impact of pre-treatment with Trichoderma asperellum and Trichoderma virens on post-harvest losses during storage and to evaluate factors associated with these losses. The storage behaviors of Allium cepa L. bulbs after different field treatments with Trichoderma spp. were examined using a complete randomized design. The common onion bulb diseases encountered during storage were basal rot and black mould diseases, with Fusarium sp., Mucor sp., Penicillium sp., Aspergillus niger and Aspergillus flavus being associated with diseased bulbs. The results regarding the percentage of diseased bulbs showed that pre-harvest treatments with Trichoderma asperellum and Trichoderma virens did not have a significant effect on disease control during storage. Furthermore, storage losses of onions were attributed to sprouting, rooting and wilting, which were exacerbated by the prevailing temperature (~30-32°C) and relative humidity (~77%-79%) during the storage period. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is a major aflatoxin B1, posing significant health concerns to humans, crops, and producer fungi. Due to the undesirable consequences of the usage of synthetic fungicides, biological control using yeasts has gained more attention. In this study, eight isolates of epiphytic yeasts belonging to Moesziomyces sp., Meyerozyma sp. and Metschnikowia sp., which have been identified as antagonists, were isolated from different plants, including grapes, blueberries, hawthorns, hoşkıran, beans and grape leaf. While volatile organic compounds (VOCs) produced by Moesziomyces bullatus DN-FY, Metschnikowia aff. pulcherrima DN-MP and Metschnikowia aff. pulcherrima 32-AMM reduced in vitro A. flavus mycelial growth and sporulation, only VOCs produced by Metschnikowia aff. fructicola 1-UDM were found to be effective at reducing in vitro AFB1 production. All yeasts reduced the mycelial growth of A. flavus by 76–91%, while AFB1 production reduced to 1.26–10.15 ng/g and the control plates' growth was 1773 ng/g. The most effective yeast, Metschnikowia aff. Pulcherrima DN-HS, reduced Aspergillus flavus growth and aflatoxin B1 production on hazelnuts. The AFB1 content on hazelnuts reduced to 333.01 ng/g from 536.74 ng/g. To our knowledge, this is the first report of yeasts isolated from plants being tested as potential biological control agents to reduce AFB1 production on hazelnuts. [ABSTRACT FROM AUTHOR]

We review herein infections by Conidiobolus spp., Cryptococcus spp., and Aspergillus spp. in sheep and goats. Conidiobolus spp. are common causes of rhinitis in sheep and are less frequent in goats, in which Conidiobolus spp. also cause skin lesions. Cryptococcus spp. cause rhinitis, meningitis, encephalitis, and pneumonia in goats, and are rarely observed in sheep. Aspergillus spp. may cause rhinitis in goats, and pneumonia and mastitis in sheep and goats. Gross and microscopic lesions caused by these 3 fungal infections may be similar to each other. The diagnosis of these diseases must be based on gross and microscopic lesions, coupled with detection of the agent by immunohistochemical, molecular, and/or culture-based methods.

Introduction: Otomycosis is a global disease, common in the world's tropical and subtropical regions. The diagnosis is clinical, but mycological examination is required for its confirmation. There is a paucity of published data on otomycosis particularly the aetiologic agents in Nigeria. This study aims to fill in this gap by assessing the clinical presentations, risk factors and aetiologic agents of otomycosis in our setting. Methods: This was a prospective study of 126 clinically diagnosed patients and 30 controls. Mycological analysis of debris and swab samples from their external auditory canal was carried out. Results: One hundred and twenty-six patients were recruited, with 162 ear samples collected. There was mycological confirmation of otomycosis in 100 (79.4%) patients (subjects) and 127 (78.4%) specimens. The subjects' age range was 1–80 years, 30.89 ± 21.15 mean age, and 29-year median age. The peak prevalent age of 1–10 years was statistically significant, P = 0.022. The common symptoms in the subjects were itching 86 (86%), ear blockage 84 (84%) and otalgia 73 (73%). Regular ear cleaning 67 (67.0%) was the most common risk factor. Noted aetiologic agents were Aspergillus species 81 (63.8%), Candida species 42 (33.1%) and Yeast 4 (3.1%). Aspergillus flavus (40/127; 31.5%) was the most common fungus isolated. Unilateral otomycosis 73 (73%) was more common than bilateral otomycosis 27 (27%). Conclusion: Otomycosis is common in all ages, and is a commonly unilateral disease. Regular ear cleaning is the most common risk factor. A. flavus was the most common aetiologic agent in this study.

Chenlu Xiao,1,2,&ast; Dan Qiao,1,&ast; Lijuan Xiong,3,&ast; Wenjie Tian,4 Dongjiang Wang,4 Shuwen Deng,5 Jian Guo4 1Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Clinical Microbiology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People’s Republic of China; 3Department of Laboratory Medicine, Second Affiliated Hospital of Traditional Chinese Medicine of Guizhou University, Guizhou, People’s Republic of China; 4Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, People’s Republic of China; 5Department of Medical Microbiology, People’s Hospital of Suzhou New District, Suzhou, Jiangsu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jian Guo; Shuwen Deng, Email guojian1110@126.com; shuwen.deng@gmail.comBackground: Aspergillus spp. infection in immunocompromised patients results in increasing morbidity and mortality. This study investigated clinical and microbiological characteristics of aspergillosis in a Chinese tertiary teaching hospital.Methods: A total of 114 patients with aspergillosis were included over a 5-year period at Ruijin Hospital. In sum, 114 Aspergillus strains were isolated and identified at species level using matrix-assisted laser desorption ionization time-of-flight mass spectrometry, confirmed by ITS gene region and β-tubulin (BenA) gene sequencing. Sensititre YeastOne was used in vitro to test susceptibility to antifungal drugs: amphotericin B, itraconazole, voriconazole, posaconazole, isavuconazole, micafungin, anidulafungin, and caspofungin.Results: The median age of the patients was 61 (19) years, men accounted for 53.5% (n=61) of the sample, about 64% were immunocompromised, and 36% had underlying diseases. Pulmonary diseases accounted for 27.2%. Aspergillus isolates were mainly isolated from sputum (n=42, 36.8%). Antifungal therapy was administered to 106 (93.0%) patients and voriconazole (n=76, 66.7%) was the most frequently used as empirical therapy. Aspergillus fumigatus (n=69, 60.5%) was the most common species. There was a 73.7% concordance between MALDI-TOF MS and molecular identification. All Aspergillus isolates showed good susceptibility to anidulafungin and caspofungin.Conclusion: Immunocompromised patients are an at-risk population for aspergillosis, and voriconazole was used as empirical therapy in Ruijin Hospital, China. A. fumigatus was the predominant Aspergillus species causing aspergillosis, and A. flavus — as non–A. fumigatus species are increasing — the second-leading cause of aspergillosis. Anidulafungin and caspofungin were the most active in vitro against the Aspergillus isolates tested. The MALDI-TOF MS method showed good accuracy for identification of common Aspergillus spp. In vitro antifungal-susceptibility testing is crucially important for decisions on effective therapy with aspergillosis.Keywords: aspergillosis, epidemiology, Aspergillus spp., MALDI-TOF MS, molecular identification, antifungal susceptibility

Background and Aim: Scientists are interested in identifying natural antibiotic substitutes that are effective against drug-resistant pathogenic microbes and spoilage fungi to counter pathogens and reduce the major public health problem of antibiotic residues in animal products. This study aimed to evaluate the antimicrobial activity of Rumex nervosus leaves (RNL) as a medicinal herb against four bacterial and two fungal strains using absolute ethanol, 50% ethanol, and aqueous extracts. Materials and Methods: The antimicrobial activities of various RNL extracts against selected microbes were evaluated using the disk diffusion antibiotic susceptibility test, minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), minimum fungicidal concentrations, and the poisoned food technique. Results: The absolute ethanol RNL extract showed the best bacteriostatic/bactericidal activity against Salmonella Typhimurium, Escherichia coli, and Staphylococcus aureus (MIC/MBC: 0.20/0.40, 0.20/0.40, and 0.32/0.65 mg/mL, respectively). The diameter of the zone of inhibition was larger (p < 0.05) for the 100% ethanol RNL extract (8.17 mm) against Salmonella Typhimurium, the 50% ethanol-RNL extract (11.5 mm) against E. coli, and the aqueous RNL extract (14.0 mm) against S. aureus than for any other bacterial isolate. The aqueous RNL extract strongly (p < 0.0001) inhibited the mycelial growth of Aspergillus fumigatus (100%) and Aspergillus niger (81.4%) compared with the control. Conclusion: The results of this study suggest that RNL is a promising new natural antimicrobial agent for food preservation. To date, most research on the antimicrobial properties of natural herbs has been conducted in vitro, with few exceptions in vivo and intervention-based research.

COVID-19-associated pulmonary aspergillosis (CAPA) has emerged as a frequent complication in the intensive care unit (ICU). However, little is known about this life-threatening fungal superinfection in solid organ transplant recipients (SOTRs), including whether targeted anti-mold prophylaxis might be justified in this immunosuppressed population. We performed a multicentric observational retrospective study of all consecutive ICU-admitted COVID-19 SOTRs between August 1, 2020 and December 31, 2021. SOTRs receiving antifungal prophylaxis with nebulized amphotericin-B were compared with those without prophylaxis. CAPA was defined according the ECMM/ISHAM criteria. Sixty-four SOTRs were admitted to ICU for COVID-19 during the study period. One patient received antifungal prophylaxis with isavuconazole and was excluded from the analysis. Of the remaining 63 SOTRs, nineteen (30.2%) received anti-mold prophylaxis with nebulized amphotericin-B. Ten SOTRs who did not receive prophylaxis developed pulmonary mold infections (nine CAPA and one mucormycosis) compared with one who received nebulized amphotericin-B (22.7% vs 5.3%; risk ratio 0.23; 95%CI 0.032-1.68), but with no differences in survival. No severe adverse events related to nebulized amphotericin-B were recorded. SOTRs admitted to ICU with COVID-19 are at high risk for CAPA. However, nebulized amphotericin-B is safe and might reduce the incidence of CAPA in this high-risk population. A randomized clinical trial to confirm these findings is warranted. [ABSTRACT FROM AUTHOR]

The heterogeneity of the lung microbiome and its alteration are prevalently seen among chronic lung diseases patients. However, studies to date have primarily focused on the bacterial microbiome in the lung rather than fungal composition, which might play an essential role in the mechanisms of several chronic lung diseases. It is now well established that Aspergillus spp. colonies may induce various unfavorable inflammatory responses. Furthermore, bacterial microbiomes such as Pseudomonas aeruginosa provide several mechanisms that inhibit or stimulate Aspergillus spp. life cycles. In this review, we highlighted fungal and bacterial microbiome interactions in the respiratory tract, with a focus on Aspergillus spp. [ABSTRACT FROM AUTHOR]

Background. The carcinogenic, mutagenic, and teratogenic chemicals such as aflatoxin are a worldwide health problem. Aspergillus spp., responsible for most cases of aflatoxin contamination, are common in the environment and spread easily to many different types of food. The objectives of this study were to conduct a survey of fungi associated with three soil invertebrates in Taif, Saudi Arabia, identify these isolates and explore mycotoxins formation. Methods. In total, 114 fungal isolates were collected from various soil invertebrates (millipedes, Armadillidium vulgare and Porcellio laevis) in Taif, Saudi Arabia, among them, 22 isolates were identified as Aspergillus spp. based on morphological and molecular characteristics followed by both Fusarium and Penicillium. Results.The sequences of ITS 1 and ITS 4 were utilized. Using bootstrap analysis, phylogenetic tree was split into two distinct clusters. Five sub clusters were included inside the first major cluster, and their bootstrap value was 99%. While, there were two small clusters in the second major cluster. All the tested Aspergillus strains were able to have a single PCR fragment amplified using the primer AspTef. TEF-1 DNA sequence bootstrap analysis with 1,000 replicates revealed two distinct groups. Additionally, the Aspergillus isolates were grouped into two different clusters with about 65% genetic similarity using ISSR-PCR analysis. The standard polymerase chain reaction was used to effectively amplify the Aopks, afl-A and omt-A genes in aflatoxigenic Aspergillus strains. Four Aspergillus strains used in this investigation were shown to generate aflatoxin B1. While, three Aspergillus stains showed ochratoxin genes. Conclusions. In conclusion, the results indicate significant differences in the fungal community between ecoregions and soil invertebrates. Moreover, mycotoxin detection and identification among Aspergillus isolates were elucidated. This study could shed light on the risk of mycotoxin contamination along the supply chain. [ABSTRACT FROM AUTHOR]

COVID-19-associated pulmonary aspergillosis (CAPA) has emerged as a frequent complication in the intensive care unit (ICU). However, little is known about this life-threatening fungal superinfection in solid organ transplant recipients (SOTRs), including whether targeted anti-mold prophylaxis might be justified in this immunosuppressed population. We performed a multicentric observational retrospective study of all consecutive ICU-admitted COVID-19 SOTRs between August 1, 2020 and December 31, 2021. SOTRs receiving antifungal prophylaxis with nebulized amphotericin-B were compared with those without prophylaxis. CAPA was defined according the ECMM/ISHAM criteria. Sixty-four SOTRs were admitted to ICU for COVID-19 during the study period. One patient received antifungal prophylaxis with isavuconazole and was excluded from the analysis. Of the remaining 63 SOTRs, nineteen (30.2%) received anti-mold prophylaxis with nebulized amphotericin-B. Ten SOTRs who did not receive prophylaxis developed pulmonary mold infections (nine CAPA and one mucormycosis) compared with one who received nebulized amphotericin-B (22.7% vs 5.3%; risk ratio 0.23; 95%CI 0.032-1.68), but with no differences in survival. No severe adverse events related to nebulized amphotericin-B were recorded. SOTRs admitted to ICU with COVID-19 are at high risk for CAPA. However, nebulized amphotericin-B is safe and might reduce the incidence of CAPA in this high-risk population. A randomized clinical trial to confirm these findings is warranted.

Background The carcinogenic, mutagenic, and teratogenic chemicals such as aflatoxin are a worldwide health problem. Aspergillus spp., responsible for most cases of aflatoxin contamination, are common in the environment and spread easily to many different types of food. The objectives of this study were to conduct a survey of fungi associated with three soil invertebrates in Taif, Saudi Arabia, identify these isolates and explore mycotoxins formation. Methods In total, 114 fungal isolates were collected from various soil invertebrates (millipedes, Armadillidium vulgare and Porcellio laevis) in Taif, Saudi Arabia, among them, 22 isolates were identified as Aspergillus spp. based on morphological and molecular characteristics followed by both Fusarium and Penicillium. Results The sequences of ITS 1 and ITS 4 were utilized. Using bootstrap analysis, phylogenetic tree was split into two distinct clusters. Five sub clusters were included inside the first major cluster, and their bootstrap value was 99%. While, there were two small clusters in the second major cluster. All the tested Aspergillus strains were able to have a single PCR fragment amplified using the primer AspTef. TEF-1 DNA sequence bootstrap analysis with 1,000 replicates revealed two distinct groups. Additionally, the Aspergillus isolates were grouped into two different clusters with about 65% genetic similarity using ISSR-PCR analysis. The standard polymerase chain reaction was used to effectively amplify the Aopks, afl-A and omt-A genes in aflatoxigenic Aspergillus strains. Four Aspergillus strains used in this investigation were shown to generate aflatoxin B1. While, three Aspergillus stains showed ochratoxin genes. Conclusions In conclusion, the results indicate significant differences in the fungal community between ecoregions and soil invertebrates. Moreover, mycotoxin detection and identification among Aspergillus isolates were elucidated. This study could shed light on the risk of mycotoxin contamination along the supply chain.

The sanitary quality of seed is essential in agriculture. This is because pathogenic fungi compromise seed physiological quality and prevent the formation of plants in the field, which causes losses to farmers. Multispectral images technologies coupled withmachine learning algorithms can optimize the identification of healthy peanut seeds, greatly improving the sanitary quality. The objective was to verify whether multispectral images technologies and artificial intelligence tools are effective for discriminating pathogenic fungi in tropical peanut seeds. For this purpose, dry peanut seeds infected by fungi (A. flavus, A. niger, Penicillium sp., and Rhizopus sp.) were used to acquire images at differentwavelengths (365 to 970 nm). Multispectral markers of peanut seed health quality were found. The incubation period of 216 h was the one that most contributed to discriminating healthy seeds from those containing fungi through multispectral images. Texture (Percent Run), color (CIELab L*) and reflectance (490 nm) were highly effective in discriminating the sanitary quality of peanut seeds. Machine learning algorithms (LDA, MLP, RF, and SVM) demonstrated high accuracy in autonomous detection of seed health status (90 to 100%). Thus, multispectral images coupled with machine learning algorithms are effective for screening peanut seeds with superior sanitary quality. [ABSTRACT FROM AUTHOR]

Filamentous fungi have always been a matter of concern in the medical field, but nowadays, due to an increase in the risk factors and the added infections with the SARS-CoV-2 virus, they are slowly but surely emerging as a dangerous health threat worldwide. Our study aims to estimate the incidence of mold infections in central Romania, as well as assess the impact the pandemic had on them while evaluating other parameters such as age, associated bacterial and fungal infections and comorbidities. Purulent discharge and respiratory secretion specimens were collected and analyzed over a period of 10 years. A total of 68 samples tested positive for molds, with an increased number of positive samples during the pandemic. The highest number of specimens came from the outpatient department, followed by medical wards, with the most common filamentous fungus being Aspergillus section Flavi. Associated diseases included affections of the respiratory system, followed by the cardiovascular system and people who suffered from a viral infection with SARS-CoV-2, and they were mostly present in seniors. The most common associated infections were with Staphylococcus aureus and Candida nonalbicans. A statistically significant correlation was found between the association of mold infection and SARS-CoV-2 and an increase in mortality. [ABSTRACT FROM AUTHOR]

The fungal cell wall (FCW) is a dynamic structure responsible for the maintenance of cellular homeostasis, and is essential for modulating the interaction of the fungus with its environment. It is composed of proteins, lipids, pigments and polysaccharides, including chitin. Chitin synthesis is catalyzed by chitin synthases (CS), and up to eight CS-encoding genes can be found in Aspergillus species. This review discusses in detail the chitin synthesis and regulation in Aspergillus species, and how manipulation of chitin synthesis pathways can modulate fungal growth, enzyme production, virulence and susceptibility to antifungal agents. More specifically, the metabolic steps involved in chitin biosynthesis are described with an emphasis on how the initiation of chitin biosynthesis remains unknown. A description of the classification, localization and transport of CS was also made. Chitin biosynthesis is shown to underlie a complex regulatory network, with extensive cross-talks existing between the different signaling pathways. Furthermore, pathways and recently identified regulators of chitin biosynthesis during the caspofungin paradoxical effect (CPE) are described. The effect of a chitin on the mammalian immune system is also discussed. Lastly, interference with chitin biosynthesis may also be beneficial for biotechnological applications. Even after more than 30 years of research, chitin biosynthesis remains a topic of current interest in mycology. [ABSTRACT FROM AUTHOR]

Eleven samples of grapes from the Central Slovak wine-growing region were examined at the time of harvest in 2021 using the plating method and the plating method with surface disinfection. Plates were incubated aerobically at 25±1 °C in the dark for one week. Identification of fungi was based on morphological and microscopic characteristics. Of the 574 strains detected and identified in the exogenous mycobiota, the most common genus was Alternaria (90.9%), followed by Aspergillus (72.7%), Cladosporium, Penicillium, and Rhizopus (63.6%, each). The most abundant genera found by descending order were Alternaria (53.3%), Rhizopus (9.4%), Aspergillus (5.5%) and only in minor percentage by Penicillium (3%), among others. Of the 512 strains detected and identified in the endogenous mycobiota, the most common genus was Alternaria (100%), followed by Rhizopus (81.8%), Penicillium (72.7%), Aspergillus (63.6%), Cladosporium (45.5%), and Epicoccum (27.3%). The most abundant genera were Alternaria (61.1%), Cladosporium (13.1%), Aspergillus (11.7%), and Rhizopus (9%). The major fungal species isolated from the genera Aspergillus and Penicillium were Aspergillus section Nigri and Penicillium glabrum. Potentially toxigenic Aspergillus and Penicillium species were evaluated for toxinogenicity by thin-layer chromatography. Positive toxinogenity was found for all strains of Aspergillus clavatus tested and for almost all strains of Penicillium expansum and P. crustosum. Out of the two Aspergillus flavus isolates tested, none produced AFB1, AFG1, and CPA. Ochratoxigenic microfungi did not produce OTA. [ABSTRACT FROM AUTHOR]

Abstract Background The effect of Aspergillus spp. and Trichoderma harzianum isolates was evaluated against the growth of Fusarium oxysporum, F. solani, Pythium ultimum and Phytophthora citrophthora affecting peach seedlings. Results The in vitro results revealed the ability of these antagonistic in reducing the radial growth of these pathogens. The most important mycelial growth reduction was of 85.82%, recorded for F. oxysporum in confrontation with A5 of Aspergillus candidus. Aspergillus flavus A4 and A. niger A10 were the most effective against F. solani with an inhibition percent more than 60%. For P. citrophthora, A. flavus A4, A. candidus A5, A. terreus A9 and A. niger A10 inhibited the mycelia growth by more than 60%. Aspergillus nidulans A1 was the most effective against Pythium ultimum (72.07%). Trichoderma harzianum isolates T9 and T10, are the most effective with a high inhibition percent of mycelial growth. The inhibition induced after 4 days of incubation, against F. oxysporum, F. solani, P. citrophthora and Pythium ultimum by these 2 antagonists exceeded 70, 60, 70 and 80%, respectively. The in planta test showed the efficacy of antagonists tested solo against some pathogens. In fact, Bacillus subtilis improved the health status by 62.55% compared to the control inoculated with P. ultimum. Trichoderma harzianum T9 significantly reduced the root rot index by 87.5% than the control inoculated with F. solani. In the same sense, B. subtilis significantly reduced this parameter by 62.55 and 88.89% than the control inoculated with P. ultimum and P. citrophthora, respectively. Furthermore, B. subtilis (B) and Aspergillus niger A10 improved plants height than the control inoculated with Pythium ultimum by 31.52 and 40.49%, respectively. However, the combinations of antagonists (T9 + T10; A5 + A10 and B + T10) did not improve their efficacy. Conclusions The isolates T. harzianum (T9 and T10), A. candidus A5 and A. niger A10 were the most effective in vitro against Fusarium, Pythium and Phytophthora species associated with peach seedling decline. The in vivo assay showed the effectiveness of B. subtilis against P. ultimum and P. citrophthora and the potential effect of T. harzianum T9 against F. solani. Their combinations revealed to be ineffective.

En animales, el consumo de alimentos fermentados contaminados con mohos productores de micotoxinas, principalmente del género Aspergillus, permite la presencia de aflatoxinas en leche cruda, lo que desencadena una problemática de salud pública. El objetivo del presente trabajo fue detectar las concentraciones y la frecuencia de aflatoxina M1 en muestras de leche cruda de vacas, almacenada en tanques de enfriamiento en cuatro municipios del departamento de Boyacá, Colombia, durante un año, para determinar las variaciones de acuerdo con la temporada. El estudio planteado fue longitudinal, de tipo descriptivo cuantitativo. Cuatro tanques de enfriamiento fueron seleccionados a conveniencia en cuatro municipios distintos del departamento y fueron muestreados dos veces al mes durante todo un año. Se realizó un análisis de varianza (ANOVA) para determinar las diferencias estadísticas entre lasconcentraciones de la aflatoxina M1 por cada trimestre. Se encontraron diferencias estadísticas entre cada uno de los trimestres del estudio: 135 muestras arrojaron resultados positivos de un total de 384 muestras colectadas en todo el estudio. La concentración máxima de la toxina fue determinada en la época seca, con un valor de 2,4 μg/kg, mientras que el valor máximo en la época de lluvias fue de 1,4 μg/kg. Se determinó por primera vez en el departamento de Boyacá las concentraciones y variaciones estacionales de aflatoxina M1 en muestras de tanques de enfriamiento de leche, donde se hallaron las mayores concentraciones y número de casos positivos de aflatoxina M1 en los meses de época seca.

The genus Aspergillus harbors human infection-causing pathogens and is involved in the complex one-health challenge of antifungal resistance. Here, a 6-year retrospective study was conducted with Aspergillus spp. isolated from patients with invasive, chronic, and clinically suspected aspergillosis in a tertiary teaching hospital. A total of 64 Aspergillus spp. clinical isolates were investigated regarding molecular identification, biofilm, virulence in Galleria mellonella, antifungal susceptibility, and resistance to amphotericin B and azoles. Aspergillus section Fumigati (A. fumigatus sensu stricto, 62.5%) and section Flavi (A. flavus, 20.3%; A. parasiticus, 14%; and A. tamarii, 3.1%) have been identified. Aspergillus section Flavi clinical isolates were more virulent than section Fumigati clinical isolates. Furthermore, scant evidence supports a link between biofilm formation and virulence. The susceptibility of the Aspergillus spp. clinical isolates to itraconazole, posaconazole, voriconazole, and amphotericin B was evaluated. Most Aspergillus spp. clinical isolates (67.2%) had an AMB MIC value equal to or above 2 µg/mL, warning of a higher probability of therapeutic failure in the region under study. In general, the triazoles presented MIC values above the epidemiological cutoff value. The high triazole MIC values of A. fumigatus s.s. clinical isolates were investigated by sequencing the promoter region and cyp51A locus. The Cyp51A amino acid substitutions F46Y, M172V, N248T, N248K, D255E, and E427K were globally detected in 47.5% of A. fumigatus s.s. clinical isolates, and most of them are associated with high triazole MICs. Even so, the findings support voriconazole or itraconazole as the first therapeutic choice for treating Aspergillus infections. This study emphasizes the significance of continued surveillance of Aspergillus spp. infections to help overcome the gap in knowledge of the global fungal burden of infections and antifungal resistance, supporting public health initiatives.

Research background. Inulinases are used for fructooligosaccharide production and they are of interest for both scientific community and industry. Black aspergilli represent a diverse group of species that has use for enzyme production, in particular some species are known as potent inulinase producers. Finding new potential producers from the environment is as important as improving the production with known strains. Safe use of enzymes produced by aspergilli in food industry is placed ahead of their benefit for inulinase production. Experimental approach. Here we show a specific approach to finding/screening of newly isolated fungal inulinase producers that combines a newly developed screening method and an equally important assessment of the toxigenic potential of the fungus. In this study 39 black aspergilli collected from different substrates in Serbia were identified and assessed for inulinase production. Results and conclusions. The most common species were Aspergillus tubingensis (51.2%), followed by A. niger (23.1%), A. welwitschiae (23.1%) and A. uvarum (2.6%). The isolates for inulinase production were selected using a cheap and easy, fast and non-hazardous alternative inulinase screening test developed in this work. Enzymatic activity of selected inulinase-producing strains was confirmed spectrophotometrically. Since some A. niger and A. welwitschiae strains are able to produce mycotoxins ochratoxin A (OTA) and fumonisins (FB), the toxigenic potential of selected inulinase producers was assessed analytically and genetically. Fungal enzyme producer can be considered safe for use in food industry only after comparing the results of both approaches for investigating toxic potential, the direct presence of mycotoxins in the enzyme preparation (analytically) and the presence of mycotoxin gene clusters (genetically). In some strains the absence of OTA and FB production capability was molecularly confirmed by the absence of complete or critical parts of biosynthetic gene clusters, respectively. The two best inulinase producers and mycotoxin non-producers (without mycotoxin production capability as additional safety) were selected as potential candidates for further development of enzyme production. Novelty and scientific contribution. The presented innovative approach for the selection of potential fungal enzyme producer shows that only non-toxigenic fungi could be considered as useful in food industry. Although this study was done on local isolates, the approach is applicable globally.

The sanitary quality of seed is essential in agriculture. This is because pathogenic fungi compromise seed physiological quality and prevent the formation of plants in the field, which causes losses to farmers. Multispectral images technologies coupled with machine learning algorithms can optimize the identification of healthy peanut seeds, greatly improving the sanitary quality. The objective was to verify whether multispectral images technologies and artificial intelligence tools are effective for discriminating pathogenic fungi in tropical peanut seeds. For this purpose, dry peanut seeds infected by fungi (A. flavus, A. niger, Penicillium sp., and Rhizopus sp.) were used to acquire images at different wavelengths (365 to 970 nm). Multispectral markers of peanut seed health quality were found. The incubation period of 216 h was the one that most contributed to discriminating healthy seeds from those containing fungi through multispectral images. Texture (Percent Run), color (CIELab L*) and reflectance (490 nm) were highly effective in discriminating the sanitary quality of peanut seeds. Machine learning algorithms (LDA, MLP, RF, and SVM) demonstrated high accuracy in autonomous detection of seed health status (90 to 100%). Thus, multispectral images coupled with machine learning algorithms are effective for screening peanut seeds with superior sanitary quality.

Invasive aspergillosis is a life-threatening complication in patients with severe influenza and COVID-19 in intensive care units. Risk factors for the invasive aspergillosis development are transitory immunosuppression associated with severe influenza and COVID-19, as well as the use of glucocorticosteroids and immunosuppressive therapy. In the presence of risk factors, suspected clinical and radiological signs of invasive aspergillosis, bronchoscopy and examination of material from the lower respiratory tract are necessary: test for galactomannan, microscopy with white calcofluor staining and inoculation on Sabouraud agar medium. Voriconazole or are recommended as first-line treatment for invasive aspergillosis in patients with severe influenza and COVID-19. Amphotericin B Liposomal, Amphotericin B Lipid Complex, and Caspofungin are the alternative options for the invasive aspergillosis treatment. Combination therapy is possible. It is necessary to control the underlying disease with eliminate or reduce the severity of risk factors.

Background and Aim: Scientists are interested in identifying natural antibiotic substitutes that are effective against drugresistant pathogenic microbes and spoilage fungi to counter pathogens and reduce the major public health problem of antibiotic residues in animal products. This study aimed to evaluate the antimicrobial activity of Rumex nervosus leaves (RNL) as a medicinal herb against four bacterial and two fungal strains using absolute ethanol, 50% ethanol, and aqueous extracts. Materials and Methods: The antimicrobial activities of various RNL extracts against selected microbes were evaluated using the disk diffusion antibiotic susceptibility test, minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), minimum fungicidal concentrations, and the poisoned food technique. Results: The absolute ethanol RNL extract showed the best bacteriostatic/bactericidal activity against Salmonella Typhimurium, Escherichia coli, and Staphylococcus aureus (MIC/MBC: 0.20/0.40, 0.20/0.40, and 0.32/0.65 mg/mL, respectively). The diameter of the zone of inhibition was larger (p < 0.05) for the 100% ethanol RNL extract (8.17 mm) against Salmonella Typhimurium, the 50% ethanol-RNL extract (11.5 mm) against E. coli, and the aqueous RNL extract (14.0 mm) against S. aureus than for any other bacterial isolate. The aqueous RNL extract strongly (p < 0.0001) inhibited the mycelial growth of Aspergillus fumigatus (100%) and Aspergillus niger (81.4%) compared with the control. Conclusion: The results of this study suggest that RNL is a promising new natural antimicrobial agent for food preservation. To date, most research on the antimicrobial properties of natural herbs has been conducted in vitro, with few exceptions in vivo and intervention-based research. [ABSTRACT FROM AUTHOR]

The antifungal resistance in non-fumigatus Aspergillus spp., as well as Aspergillus fumigatus, poses a major therapeutic challenge which affects the entire healthcare community. Mutation occurrence of cyp51 gene paralogs is the major cause of azole resistance in Aspergillus spp. To obtain a full map of genomic changes, an accurate scan of the entire length of the Aspergillus genome is necessary. In this study, using whole genome sequencing (WGS) technique, we evaluated the mutation in cyp51A, cyp51B, Cdr1B, AtrR, Hmg1, HapE and FfmA genes in different clinical isolates of Aspergillus fumigatus, Aspergillus niger, Aspergillus tubingensis, Aspergillus welwitschiae and Aspergillus terreus which responded to minimum inhibitory concentrations of itraconazole above 16 µg mL−1. We found different nonsynonymous mutations in the cyp51A, cyp51B, Cdr1B, AtrR, Hmg1, HapE and FfmA gene loci. According to our findings, Aspergillus species isolated from different parts of the world may represent different pattern of resistance mechanisms which may be revealed by WGS. [ABSTRACT FROM AUTHOR]

Traditional dry-cured hams are easily contaminated by toxigenic fungi during the fermentation and ripening stages. The detection and positive rates of ochratoxin A (OTA) are the highest among mycotoxins detected in traditional dry-cured hams, indicating that OTA in hams is a potential safety hazard to human health. This review addresses the mycotoxin-producing fungal species, the toxigenic conditions causing OTA contamination worldwide, the prevalence of OTA contamination in dry-cured hams, and the detection methods applied in OTA analysis. Additionally, this study introduces methods to prevent and control OTA in traditional dry-cured hams. The growth of common mycotoxin-producing fungi and the accumulation of mycotoxins in dry-cured ham can be controlled by a microbial starter. This review provides an important theoretical foundation for the research and control of OTA in traditional dry-cured hams. [ABSTRACT FROM AUTHOR]