Aspergillus Species Research Articles

Molecular Characterization of Fungal Isolates of Locally Processed Rice from Five Agro-Ecological Zones of Nigeria

The purpose of this study was to characterize fungal isolates contaminating locally processed rice in Nigeria and to assess the genetic similarity in the fungi isolates. Fungal species were characterized by amplification of a segment of the β-tubulin gene using Bt2aF (5’-GGTAACCAAATCGGTGCTGCTTTC-3’) and Bt2bR (5’ACCCTCAGTGT GT GACCCTTGGC-3’) primers and phylogenetic relationship established using the Maximum Likelihood method. Amplification of between 450 and 550bp of the β-tubulin region of the genomic DNA of the fungal isolates was obtained from thirty-eight samples (89%). Members of Aspergillus section flavi and section nigri groups were differentiated as A. flavus, A. parasiticus, A. tubigensis, A. sydowii and A. niger respectively. Molecular method identified Paecilomycesformosus with 99% similarity index while all the morphologically identified Penicillium species were identified to genus level as P. formosus, P. steckii and P. citrinum with a percentage similarity ranging from 95%-100%. Collectotrichum fruticola and Talaromyces flavus from SGS and NGS zone respectively were documented for the first time in the locally processed rice with 100% and 89% similarity index respectively. All the Aspergillus species were closely related to one another and narrowly diverged from the Penicillium species. Aspergilla and Penicillia were the predominant genera identified and the isolates were genetically similar irrespective of their agroecological zones.

Successful treatment of multiple intracerebral aspergillosis with voriconazole alone in an Egyptian diabetic patient with autoimmune hemolytic anemia

Aspergillosis is a common fungus that lives in soil and decaying vegetation. Inhalation of the spores causes infection mostly in immunocompromised patients. Invasive aspergillosis has an extremely high mortality, and a definitive diagnosis requires histopathological evidence of deep tissue invasion or positive culture; however, this evidence is often difficult to obtain due to the critical nature of the patients in these situations. The sensitivity of culture in this setting is also low. The galactomannan test is a recent antigen–antibody serologic test that depends on detecting an antigen which is a molecule found in the cell wall of aspergillus species. A positive result supports the diagnosis of invasive aspergillosis. We present a case of multiple intracerebral brain abscesses in an immunosuppressed patient due to an aspergillus species diagnosed by the galactomannan test with an excellent response to the treatment with the novel voriconazole alone, without any surgical intervention, and the purpose is to alert the physicians, neurologists, and infectious disease specialists to consider the intracranial aspergillosis among the differential diagnosis of the acute onset encephalitis especially in immunocompromised patients as early diagnosis and treatment may be life-saving.

The Coincidence of Aspergillus and Primary Lung Adenocarcinoma. A Case Report and Literature Review

"Although infrequent, lung cancer can be disguised by the synchronous presence of a fungal infection, most notably by the Aspergillus species. Some cases of incidental adenocarcinoma diagnosed during surgical treatment of pulmonary aspergillosis have been reported, showing just how these fungi can hide more serious, underlying pathology. This review aims to raise awareness about this occurrence to accelerate the correct etiological diagnosis, establish a timely oncological treatment, and improve the overall morbidity and mortality in lung cancer patients. In addition, we present a rather alluring case of concurrent adenocarcinoma and Aspergillus infection to display how these two entities can appear together."

Fungi involved in rhinosinusitis in arid regions: insights from molecular identification and antifungal susceptibility.

Fungal rhinosinusitis (FRS) is a common problem worldwide, with an increasing burden in arid climate regions. Aspergillus species are the most common causative agents involved. In the present study, we investigated the prevalence, molecular characterization, and antifungal susceptibility of opportunists causing FRS in Sudan on the basis of strains collected over a period of 5 years. β-Tubulin and calmodulin sequencing were used for species identification, and antifungal susceptibility profiles were evaluated by the protocol of the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Predominant species belonged to the Aspergillus flavus complex (n = 244), A. terreus complex (n = 16), A. fumigatus complex (n = 7), and other fungi (n = 17). Molecular identification of 94 strains of Aspergillus revealed the following species: A. flavus (n = 88), A. terreus (n = 1), A. citrinoterreus (n = 2), A. fumigatus (n = 1), A. caespitosus (n = 1), and A. sydowii (n = 1). Several A. flavus and an A. fumigatus isolates showed reduced susceptibility to azoles (minimum inhibitory concentrations above the clinical breakpoints or epidemiological cutoff values). Despite several mutations revealed in cyp51A of these isolates, none could be directly linked to azole resistance. Molecular identification of fungi causing FRS is useful to identify cryptic species and for epidemiologic studies. IMPORTANCE Fungal rhinosinusitis (FRS) is a significant clinical problem in arid regions. This study provides new insights into the prevalence, etiology, and antifungal susceptibility of FRS pathogens in Sudan, where the disease burden is high. Aspergillus species, particularly the A. flavus complex, were identified as the primary FRS pathogens in the region, with some evidence of antifungal resistance. The molecular identification of fungal species causing FRS is useful for detecting antifungal resistance, identifying cryptic species, and characterizing the epidemiology of the disease. The emergence of Azole resistance Aspergilli in Sudan highlights the need for continued surveillance and appropriate use of antifungal agents. These findings have important implications for clinical management, public health policy, and future research on FRS. Publishing this study in Microbiology Spectrum would enable other researchers and clinicians to build on these findings, ultimately improving the diagnosis, treatment, and prevention of FRS.

Rapid Point-of-Care Identification of Aspergillus Species in Microbial Keratitis

Microbial keratitis (MK) is a common cause of unilateral visual impairment, blindness, and eye loss in low-income and middle-income countries. There is an urgent need to develop and implement rapid and simple point-of-care diagnostics for MK to increase the likelihood of good outcomes. To evaluate the diagnostic performance of the Aspergillus-specific lateral-flow device (AspLFD) to identify Aspergillus species causing MK in corneal scrape and corneal swab samples of patients presenting with microbial keratitis. This diagnostic study was conducted between May 2022 and January 2023 at the corneal clinic of Aravind Eye Hospital in Madurai, Tamil Nadu, India. All study participants were recruited during their first presentation to the clinic. Patients aged 15 years or older met the eligibility criteria if they were attending their first appointment, had a corneal ulcer that was suggestive of a bacterial or fungal infection, and were about to undergo diagnostic scrape and culture. Sensitivity and specificity of the AspLFD with corneal samples collected from patients with MK. During routine diagnostic scraping, a minimally invasive corneal swab and an additional corneal scrape were collected and transferred to aliquots of sample buffer and analyzed by lateral-flow device (LFD) if the patient met the inclusion criteria. Photographs of devices were taken with a smartphone and analyzed using a ratiometric approach, which was developed for this study. The AspLFD results were compared with culture reports. The 198 participants who met the inclusion criteria had a mean (range) age of 51 (15-85) years and included 126 males (63.6%). Overall, 35 of 198 participants with corneal scrape (17.7%) and 17 of 40 participants with swab samples (42.5%) had positive culture results for Aspergillus species. Ratiometric analysis results for the scrape samples found that the AspLFD achieved high sensitivity (0.89; 95% CI, 0.74-0.95), high negative predictive value (0.97; 95% CI, 0.94-0.99), low negative likelihood ratio (0.12; 95% CI, 0.05-0.30), and an accuracy of 0.94 (95% CI, 0.90-0.97). Ratiometric analysis results for the swab samples showed that the AspLFD had high sensitivity (0.94; 95% CI, 0.73-1.00), high negative predictive value (0.95; 95% CI, 0.76-1.00), low negative likelihood ratio (0.07; 95% CI, 0.01-0.48), and an accuracy of 0.88 (95% CI, 0.73-0.96). Results of this diagnostic study suggest that AspLFD along with the ratiometric analysis of LFDs developed for this study has high diagnostic accuracy in identifying Aspergillus species from corneal scrapes and swabs. This technology is an important step toward the provision of point-of-care diagnostics for MK and could inform the clinical management strategy.

The Influence of Medium Composition on EUCAST and Etest Antifungal Susceptibility Testing.

There is an ongoing effort to optimize and revise antifungal susceptibility testing (AFST) methods due to the rising number of fungal infections and drug-resistant fungi. The rising antifungal resistance within Candida and Aspergillus species, which are common contributors to invasive fungal infections (IFIs), is a cause for concern, prompting an expanding integration of in vitro AFST to guide clinical decisions. To improve the relevance of in vitro AFST results to therapy outcomes, influential factors should be taken into account. The tested medium is one of several factors that could affect the results of AFST. The present study evaluated the effect of two complex media (Sabouraud dextrose and Columbia) versus the standard defined medium (RPMI 1640) on the AFST results of amphotericin B, posaconazole, and voriconazole against Candida spp. and Aspergillus spp. representatives, utilizing the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Etest methods. Overall, Candida species exhibited higher variability in minimum inhibitory concentration (MIC) across different media (more than three log2 dilutions) comparing to Aspergillus spp., while quality control isolates showed consistency regardless of tested media, antifungals, and methods. When comparing tested methods, MIC variation was mostly detected using EUCAST than it was using Etest.

Peptides from Marine-Derived Fungi: Chemistry and Biological Activities.

Marine natural products are well-recognized as potential resources to fill the pipeline of drug leads to enter the pharmaceutical industry. In this circumstance, marine-derived fungi are one of the unique sources of bioactive secondary metabolites due to their capacity to produce diverse polyketides and peptides with unique structures and diverse biological activities. The present review covers the peptides from marine-derived fungi reported from the literature published from January 1991 to June 2023, and various scientific databases, including Elsevier, ACS publications, Taylor and Francis, Wiley Online Library, MDPI, Springer, Thieme, Bentham, ProQuest, and the Marine Pharmacology website, are used for a literature search. This review focuses on chemical characteristics, sources, and biological and pharmacological activities of 366 marine fungal peptides belonging to various classes, such as linear, cyclic, and depsipeptides. Among 30 marine-derived fungal genera, isolated from marine macro-organisms such as marine algae, sponges, coral, and mangrove plants, as well as deep sea sediments, species of Aspergillus were found to produce the highest number of peptides (174 peptides), followed by Penicillium (23 peptides), Acremonium (22 peptides), Eurotium (18 peptides), Trichoderma (18 peptides), Simplicillium (17 peptides), and Beauveria (12 peptides). The cytotoxic activity against a broad spectrum of human cancer cell lines was the predominant biological activity of the reported marine peptides (32%), whereas antibacterial, antifungal, antiviral, anti-inflammatory, and various enzyme inhibition activities ranged from 7% to 20%. In the first part of this review, the chemistry of marine peptides is discussed and followed by their biological activity.

Phase-Dependent Differential In Vitro and Ex Vivo Susceptibility of Aspergillus flavus and Fusarium keratoplasticum to Azole Antifungals.

Fungal keratitis (FK) is an invasive infection of the cornea primarily associated with Aspergillus and Fusarium species. FK is treated empirically with a limited selection of topical antifungals with varying levels of success. Though clinical infections are typically characterized by a dense network of mature mycelium, traditional models used to test antifungal susceptibility of FK isolates exclusively evaluate susceptibility in fungal cultures derived from asexual spores known as conidia. The purpose of this study was to characterize differences in fungal response when topical antifungal treatment is initiated at progressive phases of fungal development. We compared the efficacy of voriconazole and luliconazole against in vitro cultures of A. flavus and F. keratoplasticum at 0, 24, and 48 h of fungal development. A porcine cadaver corneal model was used to compare antifungal efficacy of voriconazole and luliconazole in ex vivo tissue cultures of A. flavus and F. keratoplasticum at 0, 24, and 48 h of fungal development. Our results demonstrate phase-dependent susceptibility of both A. flavus and F. keratoplasticum to both azoles in vitro as well as ex vivo. We conclude that traditional antifungal susceptibility testing with conidial suspensions does not correlate with fungal susceptibility in cultures of a more advanced developmental phase. A revised method of antifungal susceptibility testing that evaluates hyphal susceptibility may better predict fungal response in the clinical setting where treatment is often delayed until days after the initial insult.

Seasonal Influence on the Microbiology and Physicochemistry of Crude Oil Polluted Wetlands

Pollutants such as crude oil spills in an environment, affect the physical, chemical and biological properties of that environment. This study was conducted to determine the effect of seasonal variations on the microbiological and physicochemical characteristics of three crude oil-polluted wetlands in Rivers State, Nigeria. Soil samples were collected at 3 depths (0-15cm, 15-30cm and 30-45cm) with the aid of a hand auger, sampling was done for a period of twelve months covering the rainy (March to August) and dry seasons (September to February). Samples were analyzed for physicochemical characteristics, using standard analytical and microbiological methods. The data obtained was analyzed using the statistical package for social science (SPSS) version 22 and Duncan’s multiple range test was used to separate means where differences occurred. Microbiological analyses showed that significantly higher microbial counts were observed in the dry season than in the rainy season in the wetland samples which could be due to favourable environmental conditions during the period of study. Wetland soil total heterotrophic bacterial (THB) count in the wet season ranged from 1.09x107cfu/g to 10.45x107cfu/g and from 2.43x107cfu/g to 18.56x107cfu/g in the dry season. Total heterotrophic bacteria had the highest population in all soil depths (0-15cm, 15-30cm and 30-45cm) in both seasons. Hydrocarbon-utilizing bacteria such as Bacillus subtilllis, B. rigui, B. flexus, Lysinibacillus macrolides, Staphylococcus aureus, Proteus penneri and fungi such as Aspergillus species, Fusarium species, Mucor species and Rhizopus species were genetically identified in this study. Temperature and other physicochemical parameters in the wetlands were altered due to the variation in season. Higher temperature ranges were observed in the dry season than in the rainy season as seen in Iwofe, temperature ranged from 26.500C (0-15cm) to 28.500C (30-45cm) in the rainy season while in the dry season it ranged from 31.500C (0-15cm) to 33.500C (30-45cm) which was above FEPA permissible limit of 300C for soil temperature. The study showed that season of study as well as crude oil spillage have significant effects on the microbiology and physicochemical characteristics of wetlands and therefore, contribute to microbiological degradation in wetlands.

Biological reduction of aflatoxin M1 in dairy products using probiotic strains: a systematic review and meta-analysis

Abstract Aflatoxins are naturally occurring toxic substances produced mainly by species of the genus Aspergillus that can contaminate almost all foodstuffs. Apart from the harmful effects they have on human and animal health, they can be secreted unchanged in animal milk and cause contamination of milk and its products. Aflatoxin M1 (AFM1) is the major and most toxic type of aflatoxin after aflatoxin B1 (AFB1). The use of probiotic strains to reduce the amount of aflatoxin in milk and by-products has been observed in many studies. In this systematic review and meta-analysis, articles in PubMed, Scopus, Google Scholar and ISI Web of Science were searched to find eligible studies that reported reduction of AFM1 using probiotics in dairy products. The results were pooled using a random-effects model. In most studies, the efficiency of probiotics in milk has been tested by Lactobacillus strains. The results indicated that probiotic microorganisms could significantly reduce AFM1 by 55.76% (confidence interval (CI): 54.35%, 57.16%; I2 = 100%). Besides, the rank order of AFM1 reduction in dairy products based on probiotic strain subgroup was: Lactobacillus 51.99% (CI: 50.14%, 53.85%, I2 = 100%), Saccharomyces 67.36% (CI: 65.05%, 69.67%, I2 = 100%), Bifidobacterium 54.80% (CI: 54.18%, 55.43%, I2 = 99.9%), and 61.90% (CI: 53.80%, 70.00%, I2 = 100%) by a mix of strains. Considering the high binding potential of AFM1 to probiotic strains, these microorganisms can be recommended as a safe system to reduce AFM1 in dairy products.

Filters of automobile air conditioning systems as in-car source of exposure to infections and toxic moulds

The main component of an air conditioning system is air filters. Over time, the filters of an air conditioning system in cars can turn into sources of emission of microbiological hazards. The aim of this study was to quantitatively and qualitatively assess the presence of infectious and toxic fungi in the AC filters in passenger cars. The studied non-woven filters were removed from passenger cars during the “winter”/“summer” seasons. The taxonomic identification of the fungi isolated from the filters was performed using both the culture-based and molecular methods. RT-PCR was applied to assess the presence of gene fragments regulating aflatoxin biosynthesis in the isolates obtained from fungal cultures. The average fungal concentrations in the filter samples collected during the summer/winter season were 5.4 × 104 cfu/m2 and 2.4 × 104 cfu/m2, respectively. Most of the filter samples, collected in both the studied seasons, revealed the presence of Aspergillus species including A. fumigatus, A. niger, A. terreus and/or A. flavus. The recorded levels of fungal contamination of AC filters in passenger cars indicate the necessity for more frequent filter replacement in this type of vehicle. Occupational exposure to moulds and the resulting health problems that may be experienced by professional drivers should be properly recognised in order to undertake effective preventive measures.

Evaluation of the Antibiotic Susceptibility Patterns of Isolates from Feed and Water of Selected Poultry Farms in Awka Anambra State, Nigeria

The susceptibility of microorganisms isolated from poultry feeds and poultry water samples to selected antibiotics was assessed. Standard methods were used to analyze selected poultry feeds and poultry water samples. The antibiotic susceptibility patterns of the bacterial isolates were determined against the following antibacterial agents: erythromycin (10 µg), ciprofloxacin (10 µg), ampiclox (20 µg), rifampicin (20 µg), amoxil (20 µg), septrin (30 µg), ampicillin (30 µg), ceporex (10 µg), levofloxacin (20 µg), gentamicin (10 µg), streptomycin (30 µg), norfloxacin (10 µg), chloramphenicol (30 µg), ofloxacin (10 µg), nalidixic acid (30 µg), reflecine (10 µg), and augmentin (30 µg).The highest viable counts of bacteria isolated from poultry feed and water samples were 2.7x106 cfu/g and 1.69x103 cfu/ml, respectively. The highest fungal counts in the poultry feed and water samples were 1.60x105 cfu/g and 2x105 cfu/ml, respectively. Bacterial isolates from poultry feed and water samples included Escherichia coli, Klebsiella pneumoniae, Salmonella species, and Staphylococcus aureus. Fungal isolates included Aspergillus species, Penicillium species, Mucor species, and Candida species.Staphylococcus aureus exhibited the highest susceptibility to most of the antibiotics, while Klebsiella pneumoniae showed the highest resistance, as it was resistant to five out of the ten antibiotics tested in this study. The research has demonstrated that poultry feed and poultry water showed varying levels of contamination, which may pose serious health risks to poultry. Amoxil, levofloxacin, ciprofloxacin, reflecine, and ofloxacin are recommended for use as antibiotics to treat diseases that may be caused by some of these pathogens.

Regional distribution and diversity of Aspergillus and Penicillium species on Croatian traditional meat products

Various factors, such as weather and production practices (e.g., environmental hygiene, process duration, raw material quality, ripening temperature, and relative humidity), in combination with the intrinsic product properties (e.g., pH, aw, salt content), significantly affect the growth of surface moulds. The aim of this study was to isolate and identify surface moulds retrieved from traditional meat products (TMPs) and correlate these data to the production region and production technology. The surface of 250 TMPs (dry-fermented sausages, n = 108; dry-cured meat products, n = 142) from five Croatian regions were sampled during a two-year period. Dry-fermented sausages had a significantly higher pH and a lower salt concentration when compared to dry-cured meat products. In total, 528 isolates were obtained, comprising 20 Penicillium and 17 Aspergillus species. The species most frequently isolated from the dry-fermented sausages were P. commune (32.4 %), A. proliferans (33 %), and P. solitum (14.8 %), while A. proliferans (52.1 %), P. commune (28.9 %) and P. citrinum (19.7 %) predominated in dry-cured meat products. Aspergillus predominated on the TMPs from southern Croatia, while Penicillium was prevalent on products from the other four regions, possibly due to differences in weather conditions. Seven potentially mycotoxigenic species (A. creber, A. flavus, A. niger, A. westerdijkiae, P. citrinum, P. commune, and P. nordicum) were isolated and identified. Regular monitoring of mould species and their toxigenic metabolites present on traditional meat products is of the utmost importance from the public health perspective, while the results of such a monitoring can prove beneficial for the tailoring of the production technology development.

Biochemical characterization of an acid-thermostable glucoamylase from Aspergillus japonicus with potential application in the paper bio-deinking.

Aspergillus species have been highlighted in enzyme production looking for industrial applications, notably, amylases are one of the most interesting enzymes. They are capable of hydrolyzing α-glycosidic linkages of starch and widely used in industrial processes to produce ethanol, glucose, and fructose syrup as well as in the textiles, detergents, and paper industries applications. In this context, this work aimed at the biochemical characterization of the glucoamylase from Aspergillus japonicus and its application in the bio-bleaching process of recycled paper. The optimum temperature and pH for the glucoamylase assay were standardized as 50°C and 5.5. After 1 h of incubation, glucoamylase retained 90% of its activity at 30-50°C. It also kept 70% of its activity in the pH range of 4.0-6.5 after an hour of incubation. The enzyme led to an increase of 30% in the relative whiteness of 10 dry grams of sulfite paper and magazine paper when applied along with commercial cellulase and 10 mM MnCl2 . In addition, after the treatments, the glucoamylase recovered activity was 30%-32%, which indicates a prolonged availability of the enzyme and can considerably curtail the redundant downstream process of the recycled paper bio-bleaching. Thus, the glucoamylase from A. japonicus has a significant role in bio-bleaching recycled paper, reducing the necessity of hard chemicals, and improving the industrial process in an interesting economic and ecological mode.

Accelerated Degradation of four Organophosphorus Insecticides by Malathion Tolerant Aspergillus niger MRU01 a Soil Fungus

Insecticides are widely used in agriculture, horticulture, medicine, and industry, as well as domestically. Increased insecticide application has been attributed to higher agricultural productivity and lower infield and postharvest crop loss in the twentieth century. Malathion is a very commonly used organophosphorus (OP) insecticide applied in over 100 food crops in agriculture and post-harvest storage. Different species of Aspergillus have shown their efficiency to degrade OP chemicals. A malathion tolerant Aspergillus niger MRU01, developed by prolonged malathion exposure, was tested for its efficiency to remove malathion as well as three other commonly used OP insecticides, viz., parathion, chlorpyrifos and dimethoate. The tolerant strain showed growth efficiency as well as enhanced production of phosphatases and esterase enzyme as compared to untreated and treated wild type (A. niger ATCC16404). The removal of malathion was 29% and 68%, in the wild type and the tolerant strain, respectively during five days. The tolerant strain was also able to remove dimethoate (63%) and chlorpyrifos (54%) with high efficiency while a low rate of removal of parathion (33%) was observed. The strain can be tested in the microcosms more specifically for bioremediation of other OP insecticides before its application in the field.

Haematopoietic Stem Cell Transplantation for Chronic Granulomatous Disease.

Chronic granulomatous disease (CGD) is an inborn error of immunity due to defects in the transport or function of subunits of nicotinamide adenine dinucleotide phosphate oxidase, the enzyme that generates the phagocyte respiratory burst responsible for intracellular killing of engulfed micro-organisms. Patients present with infectious or inflammatory complications. Common bacterial pathogens include Staphylococcus aureus and Burkholderia cepacia complex. Fungal pathogens include Aspergillus species, particularly Aspergillus fumigatus. Inflammatory complications most commonly manifest as inflammatory bowel disease or lung disease. Granulomata are the distinguishing histological feature. Haematopoietic stem cell transplantation (HSCT) was first considered for CGD in the early 1970's. Since then, refinements in transplant technique, donor selection, conditioning regimens, and graft engineering have widened the option of HSCT to most patients with CGD. This review charts the progress made in HSCT for CGD.

МИКОБИОТА В ДОННЫХ ГРУНТАХ ГОРЯЧИНСКОГО ТЕРМАЛЬНОГО ИСТОЧНИКА

The number of colony-forming units (CFU), composition and taxonomic structure of fungal biota in the bottom grounds of the Goryachinsk geothermal spring (Buryatia, Russian Federation) were determined by the cultural approach and the method of high-performance sequencing. The number of fungi in the grounds (0–3 cm) at the outlet of the spring and the watercourse at a short distance (3–100 m) varied in the range from several units to hundreds of CFU in 1 g. 70 isolates of fungi of 34 morphotypes were isolated by platings on nutrient media, of which 15 were identified to the species and 2 to the genus level. Its were thermophilic and thermotolerant species Aspergillus fumigatus, A. niger, A. nishimurae, A. terreus, Melanocarpus albomyces, Mycothermus thermophilus, Naganishia brisbanensis, Penicillium sp., Pseudothielavia terricola, Rhodotorula sp., Scedosporium apiospermum, Talaromyces flavus, Thermomyces dupontii, Thermothielavioides terrestris and Vishniacozyma carnescens. The method of high-performance sequencing of the ITS2 rDNA site in the source soils revealed an order of magnitude greater diversity of fungi, 149 species of 132 genera, and a total of 160 species were detected by both approaches. The number of identified operational-taxonomic units (OTE) to the species was 64.4%. In addition to ascomycetes and basidiomycetes, which were detected by platings on nutrient media, DNA barcoding showed the presence of representatives of the Phylums Mucoromycota, Mortierellomycota, Zoopagomycota, Chytridiomycota, Rozellomycota in the grounds. Moreover, among the fungi established by high-performance sequencing were not only thermotolerants, but also species with different resistance to high temperatures and trophic orientation. The application of both approaches provided more detailed information about the diversity of fungal organisms in the hot spring. However, in order to identify the inhabitant species of such ecotopes, a thorough analysis of their physiological and biochemical properties (which for many taxa is absent in due to volume) and the use of other approaches are necessary.

Highly Effective and Broad-Spectrum Antimicrobial Quaternary Ammonium Salts Containing Camphene Structure: Preparation, Surface-Active Properties, and Bioassay.

A series of novel quaternary ammonium salts (QASs) (4a-4n) comprising a camphene moiety were synthesized for the first time. Fourteen examples were prepared from camphene through Prins reaction, halogenation, and quaternarization, successively. The structures of the synthesized QASs were analyzed by Fourier transform infrared spectroscopy, 1H NMR, 13C NMR and high-resolution mass spectrometry. Surface-active properties, emulsifying abilities, and foaming properties of the investigated compounds were then studied. The antimicrobial activities of these QASs against Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae), and fungi species (Candida albicans, Candida tropicalis, and Aspergillus niger) were determined by the microdilution method. The results showed that the chemical structural factors, including types of substitutes and alkyl chain length, might be correlated with the lipid-water partition coefficient (cLog P), which played a critical role in the antimicrobial process. Compounds with alkyl chain lengths (N) in the range of 10-14 were relatively more active, while compounds bearing pyridinium, benzyl, methylimidazolium groups, or varied alkyl chain lengths (N < 5 and N > 16) were almost inactive. Compound 4k possessing a dodecyl group exhibited the most effective and broad-spectrum antimicrobial activity against almost all tested bacteria and fungi with the minimal inhibitory concentration values ranging from 0.24 to 0.98 μg/mL.

Isolation and Identification of Microfungi from Diesel Oil Contaminated Soils and Their Physico-Chemical Analysis

Diesel oil contaminated soils were collected and analyzed for various Physico-chemical properties that included moisture content, total carbon, total nitrogen and phosphorus using standard protocol and the data compared from normal uncontaminated soils. Analysis revealed that all 5 samples of diesel oil contaminated soils possessed higher percent of moisture content, greater amounts of total carbon, nitrogen and phosphorus in comparison to non-contaminated control soils. The diesel oil contaminated soils are not suitable for cultivation of agricultural crops. Though, several species of fungi were isolated from diesel contaminated soils but only 6 species of Aspergillus utilized diesel oil as sole carbon and nitrogen source as these grew well and sporulated normally on 2% agar that contained 10% of diesel oil only as nutrient. These included Aspergillus luchuensis, Aspergillus violaceofuscus, Aspergillus niveus, Aspergillus terreus, Aspergillus japonicus, Aspergillus ustus. which also grew well in broth containing 0.5% jaggery (for initial growth) supplemented with 10% diesel oil only which suggest that these species could utilize diesel oil as sole carbon and nitrogen source because no other nutrient other than diesel oil was added into the medium. The role of carbon, nitrogen and phosphorus is soils has been discussed in this paper.

Azole-Containing Agar Plates and Antifungal Susceptibility Testing for the Detection of Azole-Resistant Aspergillus Species in Hospital Environmental Samples.

The indoor environment of hospitals should be considered as an important reservoir of azole resistant Aspergillus species. In this study, we evaluated azole-containing agar plates (ACAPs) and antifungal susceptibility testing (AFST) for the detection of azole-resistant Aspergillus species in hospital environmental samples. Between September 2021 and January 2022, environmental samples (108 instruments and 12 air) were collected from different wards of 4 educational hospitals in Mazandaran province, Iran. All samples were cultured using ACAPs. Recovered Aspergillus isolates were molecularly identified at species level using partial DNA sequencing of beta-tubulin gene. AFST of Aspergillus species was performed using the Clinical and Laboratory Standards Institute M38-A3 guideline. Screening for cyp51A mutations was also done. Overall, 18 (15.0%) isolates of Aspergillus species were recovered from ACAPs, of which Aspergillus tubingensis (50%) and Aspergillus fumigatus (38.9%) were the commonest species. No isolate of Aspergillus species grew on posaconazole (PCZ)-containing agar plates. Among the 18 Aspergillus isolated species from ACAPs, 83.3% were related to samples from instruments. Of the nine isolates of A. tubingensis, 22.2% and 44.4% isolates showed minimum inhibitory concentration (MIC) = 2 μg/mL against voriconazole (VCZ) and itraconazole, respectively; and 44.4% isolates showed MIC = 1 μg/mL against PCZ. Of the seven isolates of A. fumigatus, one (14.3%) was resistant to VCZ. This isolate showed F46Y, G54E, G138C, M172V, M220I, D255E, T289F, G432C, and G448S mutation in cyp51A. Our finding showed the emergence of high MICs in cryptic and non-fumigatus species of Aspergillus such as A. tubingensis and VCZ resistance in A. fumigatus in indoor environment of hospitals.