Mold Species Research Articles

718. Posaconazole Versus Standard of Care for Treatment of Invasive Mold Infections

Abstract Background Invasive mold infections (IMIs) including Aspergillus spp. are a significant cause of morbidity and mortality in immunocompromised patients. Voriconazole is recommended as first line treatment for primary IA. Liposomal amphotericin B and isavuconazole are also recommended alternative therapies. Posaconazole has activity against mold species including those not covered by voriconazole, is available as an IV and oral formulation with a distinct adverse effect profile making it a potential effective alternative for treatment. This study investigated the clinical outcomes of treatment of invasive mold infections (IMIs) with posaconazole compared to standard of care (voriconazole, isavuconazole, liposomal amphotericin B). Methods A retrospective, single center, cohort study was completed to evaluate patients with IMIs treated with posaconazole versus those treated with standard of care therapy. Medication orders for posaconazole or standard of care therapy between January 2012 and December 2020 were identified from a clinical data repository. Only patients with antifungal orders with a listed indication for “treatment of fungal infection” were included. Data collected for each group included baseline demographics, underlying conditions, site of infection, type of mold, therapeutic drug monitoring and classification of IMI. Results A total of 120 patients met inclusion criteria, with 35 patients in the posaconazole group and 85 patients in the SOC group. Baseline characteristics were similar except for increased severe neutropenia in the posaconazole group (p< 0.0001), more probable IMIs in the SOC group (p=0.009) and more possible IMIs in the posaconazole group (p=0.043). In the posaconazole group 37.1% (13/35) of patients were treatment experienced vs. 29.4% (25/85) of patients in the SOC group. More patients achieved a complete/partial response in the SOC group compared to the posaconazole group (p=0.0001) and more patients experienced treatment failure in the posaconazole group (p=0.01). A higher proportion of patients experienced adverse effects in the SOC group compared to the posaconazole group (p=0.0001). Conclusion Posaconazole was not as effective as SOC in treating invasive mold infections but patients experienced comparatively fewer adverse events. Disclosures All Authors: No reported disclosures

Misalignment between Clinical Mold Antigen Extracts and Airborne Molds Found in Water-damaged Homes

Rationale: Epidemiologic studies have demonstrated that exposure to molds and other fungi can play a role in a variety of allergic and pulmonary diseases in susceptible individuals. Species-specific mold antigen extracts are used in the clinical evaluation of suspected mold-related conditions; however, alignment between these extracts and the species of molds identified in the indoor environment of water-damaged homes has not been rigorously evaluated. Objectives: To identify the predominant genera and species of mold in the air of homes with water damage, mold growth, and/or occupants with respiratory complaints (complaint homes), and to assess their alignment with the mold antigen extracts used in clinical practice. Methods: The genera and species of molds identified in culture-type outdoor and indoor air samples collected from complaint homes throughout the United States and Canada from 2002 to 2017 were examined. Mold antigen extracts available and utilized for skin and serum testing in clinical practice were assessed, and alignment between these data were evaluated. Results: Culture data from 24,455 indoor air samples from 7,547 complaint homes and 29,493 outdoor samples were evaluated. Mean exposure values (colony-forming units [cfu]/m3) were calculated for each genus and species and indoor versus outdoor values were compared. Penicillium was the predominant genus identified in water-damaged homes, with a mean exposure (233.3 cfu/m3) 2.9 times higher than that of the Aspergillus genus (81.4 cfu/m3). Five Penicillium (P. aurantiogriseum, P. brevicompactum, P. citrinum, P. crustosum, and P. variabile) and three Aspergillus (A. versicolor, A. sydowii, and A. niger) species were identified as the predominant indoor water-damage-related fungi. However, none of these Penicillium species and only one of the Aspergillus species is currently available as an antigen extract for use in skin testing or serum testing panels. Conclusions: Significant misalignment exists between the currently available mold antigen extracts and the predominant species of molds found in water-damaged homes. Improving alignment has the potential to enhance diagnosis of mold-related diseases, including allergic asthma and hypersensitivity pneumonitis and to improve patient outcomes via interventions, including antigen avoidance through building remediation and occupant relocation, consistent with the findings of a recent American Thoracic Society Workshop Report.

Steroid-Functionalized Imidazolium Salts with an Extended Spectrum of Antifungal and Antibacterial Activity.

It is established that high rates of morbidity and mortality caused by fungal infections are related to the current limited number of antifungal drugs and the toxicity of these agents. Imidazolium salts as azole derivatives can be successfully used in the treatment of fungal infections in humans. Steroid-functionalized imidazolium salts were synthesized using a new, more efficient method. As a result, 20 salts were obtained with high yields, 12 of which were synthesized and characterized for the first time. They were derivatives of lithocholic acid and 3-oxo-23,24-dinorchol-4-ene-22-al and were fully characterized by 1H and 13C nuclear magnetic resonance (NMR), infrared spectroscopy (IR), and high resolution mass spectrometry (HRMS). Due to the excellent activity against bacteria and Candida albicans, new research was extended to include tests on five species of pathogenic fungi and molds: Aspergillus niger ATCC 16888, Aspergillus fumigatus ATCC 204305, Trichophyton mentagrophytes ATCC 9533, Cryptococcus neoformans ATCC 14116, and Microsporum canis ATCC 11621. The results showed that the new salts are almost universal antifungal agents and have a broad spectrum of activity against other human pathogens. To initially assess the safety of the synthesized salts, hemocompatibility with host cells and cytotoxicity were also examined. No toxicity was observed at the concentration at which the compounds were active against pathogens.

Practical Approach to the Pathologic Diagnosis of Mixed Filamentous Fungal Infections in Burn Wounds: A Case Series

Abstract Introduction/Objective Opportunistic infections by fungi are a major source of morbidity and mortality in patients suffering from extensive burn wounds. Here we review a series of cases of infections by multiple fungi in burn wounds as diagnosed by histopathology and outline the key features for the pathologist to include in the report. Methods/Case Report Biopsies from patients with more than one fungal species identified in the laboratory in a concurrent culture or by PCR were included in this study. Three cases are presented with multiple fungi identified. Each case had yeast and at least one different hyaline mold species present on pathology; two cases additionally had mucormycetes present, with angioinvasion in one case. All organisms requiried microbiologic cultures and variably required molecular testing for full identification. Results (if a Case Study enter NA) N/A Conclusion Pathologists should be aware of the possibility of infection by multiple fungal species in burn wounds. Fungal morphology in tissue sections should allow for detection and distinction of mucormyctes and other hyaline molds. Histopathologic correlation with culture and/or PCR results is essential to distinguish potential contaminants from true infection.

Emergency Response Validation of the Soleris® Direct Yeast and Mold Method for Detection of Yeast and Mold in Dried Cannabis Flower: AOAC Performance Tested MethodSM 051301.

Soleris® Direct Yeast and Mold (DYM) is a growth-based, automated method for detection of yeast and mold in select foods and other sample types including nutraceuticals and cosmetics. The Soleris method is used in a "dilute-to-specification" or threshold manner in which a result is scored as positive or negative around a predetermined cutoff (in CFU/g) established by the dilution and volume of sample homogenate tested. The objective of this study was to validate the method for testing of dried cannabis flower. The validation was conducted under the Emergency Response Validation program of the AOAC Research Institute. The study included inclusivity and exclusivity testing, in particular testing of yeast and mold species associated with cannabis, and a matrix study in which Soleris method presumptive results were compared with Soleris confirmed results using Dichloran Rose-Bengal Chloramphenicol (DRBC) agar for confirmation. Samples at four different levels of natural yeast and mold contamination were tested at two test thresholds. In inclusivity testing, all 63 yeast and mold strains tested produced positive results within the specified test duration of 72 h. In exclusivity testing, 36 of 37 strains tested produced no detection within 72 h. In matrix testing, there were no significant differences between Soleris presumptive and confirmed results for any contamination level or test threshold as determined by probability of detection analysis. Results indicate that the Soleris method is an effective procedure for detection of yeast and mold in dried cannabis flower. With the Soleris method, results are available within 72 h compared with the 5-7 days required for microbiological culture methods.

Application of dielectric barrier discharge for improving food shelf life and reducing spoilage

Dielectric Barrier Discharge (DBD) based ozone therapy is an attractive non-thermal, additive-free and environment-friendly alternative to traditional food processing technologies. Its practical application is dependent on economical ozone generation and optimum ozone dosage. This study investigates the one-time and periodic application of a compact (48 cu. cm), lightweight (55 g), low power, low temperature, DBD ozone generator for treatment of spoilage inocula prepared from combinations of spoiled green beans, grape tomatoes, lettuce and strawberries. A one-time exposure of 126–136 ppm of average ozone concentration produced by the DBD generator over 3 min and 15 min resulted in at least 1 and 4 log reduction, respectively, in microbial colonies present in the spoilage inocula. Daily exposure of 128.7 ppm average ozone concentration over 3 min under similar conditions showed that inhibition through periodic exposure can successfully inhibit the growth of both bacteria and mold species with at least 5 log reduction of microbial colonies. Visual inspection of whole fruits and vegetables with similar 3-min daily exposure showed the potential of ozone therapy to at least double the shelf-life of food products. For the daily exposures, energy required by the DBD ozone generator was calculated as 0.39 pm 0.06 kJ/day.

A Review on Mucormycosis (Black Fungus) and its Recent Study

Mucormycosis is an infrequent fungal phycomycosis disease caused by species of molds called Mucormycetes and it proliferates over India. The infection is analogous to the upper or lower airways and clinical amelioration. Besides, use abundant surgical and medical remedies. The characteristics which are responsible for mucormycosis are neutropenia, diabetic ketoacidosis, protein-calorie malnutrition, iron overload. The various types of mucormycosis should mention in the article. Also, signs and symptoms, prevention, and diagnosis are granted in an object.

A fungal antigenic driver for Löfgren's syndrome sarcoidosis.

Löfgren's syndrome is an acute form of sarcoidosis that is characterized by the activation of CD4+ T helper cells. In this issue of JEM, Greaves et al. (2021. J. Exp. Med.https://doi.org/10.1084/jem.20210785) identified a peptide derived from an airborne mold species that stimulates T cells of Löfgren's syndrome patients in an HLA-DR3-restricted manner. An increased serum IgG antibody response to the full-length protein was also observed in those patients, indicating that the fungus Aspergillus nidulans might be the elusive microbial agent that drives acute sarcoidosis.

Diversity of Filamentous Molds Producing Mycotoxins in Rice Called "Deni Kachia" Sold on the Markets of Daloa (Côte d’Ivoire)

Filamentous fungi are frequent contaminants of many plant substrates and certain animal products. Their presence can improve the organoleptic quality of the product or alter it and lead to the accumulation of toxic secondary metabolites, such as mycotoxins. The objective of this work is to characterize the filamentous fungi isolated from the rice "deni kachia" stored and sold on public markets in the city of Daloa. A count and isolation of filamentous fungi, followed by macroscopic and microscopic identification was carried out. Several genera of filamentous fungi was identified. These are the genera Aspergillus, Penicillium and others. The different species of molds identified by conventional biochemical techniques were confirmed by the API 50 CH freezer.The genera Aspergillus and Penicillium are the most predominant in the samples analyzed. The presence of these species with their ability to produce mycotoxins suggests that contaminated rice poses a risk to the health of consumers.

Three novel sooty moulds species of Trichomerium from Yunnan, China

Collection of three sexual morphic fungal species occurring on the leaf surface of three plant species (Dalbergia obtusifolia, Syzygium fluviatile and Ziziphus mauritiana) collected in Yunnan Province, China, have shown that they are morphologically distinct. Phylogenetic analyses of partial sequences of internal transcribed spacer region (ITS) and large subunit rDNA (LSU) placed the three species in Trichomerium (Trichomeriaceae) but are sufficiently different from all known species. Therefore, three new species, Trichomerium multisetosum, T. xishuangbannaense and T. yunnanense are described with detailed, illustrated descriptions and notes.

Potential of Near Infrared Spectroscopy as a Rapid Method to Discriminate OTA and Non-OTA-Producing Mould Species in a Dry-Cured Ham Model System.

The ripening process of dry-cured meat products is characterised by the development of fungi on the product’s surface. This population plays a beneficial role, but, uncontrolled moulds represent a health risk, since some of them may produce mycotoxins, such as ochratoxin A (OTA). The aim of the present work is to assess the potential of near-infrared spectroscopy (NIRS) for the detection of OTA-producing mould species on dry-cured ham-based agar. The collected spectra were used to develop Support Vector Machines–Discriminant Analysis (SVM-DA) models by a hierarchical approach. Firstly, an SVM-DA model was tested to discriminate OTA and non-OTA producers; then, two models were tested to discriminate species among the OTA producers and the non-OTA producers. OTA and non-OTA-producing moulds were discriminated with 85% sensitivity and 86% specificity in the prediction. Furthermore, the SVM-DA model could differentiate non-OTA-producing species with a 95% sensitivity and specificity. Promising results were obtained for the prediction of the four OTA-producing species tested, with a 69% and 90% sensitivity and specificity, respectively. The preliminary approach demonstrated the high potential of NIR spectroscopy, coupled with Chemometrics, to be used as a real-time automated routine monitorization of dry-cured ham surfaces.

Characterisation of probiotic properties of yeast strains isolated from kefir samples

AbstractThis study aimed to isolate and identify yeast species from kefir samples and to determine their probiotic characteristics in comparison with commercial probiotic yeast Saccharomyces boulardii. The microbial flora of traditional kefir samples were dominated by Saccharomyces cerevisiae and Kluyveromyces marxianus. Kefir isolates demonstrated higher auto‐aggregation, hydrophobicity and antioxidant properties compared to S. boulardii although S. boulardii demonstrated the highest survival rate under in␣vitro gastrointestinal digestion conditions followed by two kefir yeasts K. marxianus and Kazachstania unispora. Importantly, kefir isolates revealed strong antifungal activities against mould species except Penicillium roqueforti. These findings suggest probiotic and functional roles of yeast strains isolated from traditional kefir samples.

Japanese Traditional Miso and Koji Making.

Miso is a traditional Japanese seasoning paste produced by fermenting soybeans using the power of koji mold. A recent Japanese cohort study has shown that increased consumption of fermented soybean products is associated with a reduced risk of death in both men and women. In this review, we briefly explain what miso means in the Japanese culture and food industry, varieties of miso available today, and steps involved in miso making. Then, we review early and latest scientific researches in koji mold species, their safety, and beneficial enzymes they produce during fermentation and maturation processes, which play a major part in determining the quality and sensory profile of miso.

Myxomycete-Coleoptera associations in the Polish Carpathians

ABSTRACT Few papers have been published on the interactions between slime moulds and beetles in Europe. The available knowledge was usually acquired incidentally to mycological studies and concerns common and easily identifiable slime moulds. The beetles in those studies were frequently identified to genus only, and sometimes only selected families were studied. In 2018–2019, a comprehensive survey of Myxomycete-Coleoptera associations was carried out in the Pieniny National Park (Carpathians, S Poland). A total of 164 sporocarps from 30 species of slime moulds were examined, and beetles were found in 112 sporocarps of 23 slime mould species. The sporocarps were inhabited by 674 individuals from 37 species of beetles. To date, this is the most comprehensive study of beetles associated with slime moulds in Europe.

Contamination of Acorns of Pedunculate Oak (Quercus robur L.), as Feed Material, by Moulds and Mycotoxins

Abstract In the past, pigs were commonly fed with acorns, and this was of remarkable economic importance. Currently this habit is continued in some areas, especially for production of prime-quality Iberian ham. Mature acorns, after shedding and during storage in unsuitable conditions, can be quickly infected with spores of many moulds, which cause mummification, blackening, dehydration, and nutrient loss. This study aimed to evaluate the quality of acorns of pedunculate oak (Quercus robur L.). The samples were collected in 2017 in southern Wielkopolska (central part of Poland), as feed material. In mouldy acorns a very high number of fungi was found (2.6 × 106 cfu/g), and 97% of them represented pathogenic Penicillium spp. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed in mouldy acorns high concentrations of mycophenolic acid (14580 μg/kg) and patulin (50 μg/kg). The dominant mould species, Penicillium expansum, showed a high cytotoxicity of swine kidney cells using assay based on the conversion of the tetrazolium salt, 3-(4,5, dimethylthiazol-2-yl)-2-5 diphenyltetrazolium (MTT). This raises the question if the pathogenic metabolites of moulds present in acorns can be dangerous for livestock, especially pigs, and people, as acorns are beginning to be seen as an interesting and functional part of their diet.

Inhibitory effects of binary combinations of microbial metabolites on the growth of tolerant Penicillium roqueforti and Mucor circinelloides

Dairy product spoilage by molds leads to substantial food waste and economic losses. Although some bioprotective lactic acid bacteria exhibit antifungal activity, some spoilage molds displayed extensive tolerance to these antifungal cultures and their metabolites. This study aimed to investigate some combinations of selected microbial compounds on two molds, Penicillium roqueforti ISI4 and Mucor circinelloides 01180023, previously found to be tolerant. Of the thirteen compounds tested in susceptibility test, octanoic acid (OA) exhibited the most potent antifungal activity, followed by diacetyl and 3-Phenylpropanoic acid (3-PPA). Six compounds were chosen for assessing the antifungal activity of binary combinations. OA combined with either diacetyl (COD) or 3-PPA (COP) displayed synergistic antifungal effects in laboratory medium and yoghurt. Using oCelloScope, mold growth inhibition and morphological changes induced by COD and COP were observed. The increase of extracellular conductivity, intracellular materials leakage, and the propidium iodide-stained spores demonstrated the membrane was damaged. The accumulation of reactive oxygen species and the elevated malondialdehyde (MDA) content indicated the two combinations induced oxidative stress, leading to membrane lipid peroxidation. These findings highlight some combinations may exhibit synergistic effects and open up for opportunities to design culture and metabolite mixes with effect even towards very robust mold species.

Influence of Dietary Supplementation of Ginger, Garlic and their Combination on Mycological Assessment of Quail Carcasses

Objectives: this work was under taken to evaluate the influence of dietary supplementation of ginger, garlic and their combination on quail meat through assessment of its contamination with fungi and its toxins and its effect on human. Methods: We obtained 110, Japanese quail chicks of one day old were included in this study, they were further subdivided into four groups (n = 25/group): Control group was free from any dietary additions and the other three treated groups with ginger, mixed and garlic supplements, respectively for 45 days. After scarifying, skin swab specimens were assembled from quail breast and thigh muscles. Results: revealed that breast and thigh muscles in garlic treated group have the lowest counts of total mold and yeast than those of the others. Notably, control group represented the highest incidence in mold and yeast contamination as compared to that of the others. The isolated mold species were Aspergillus spp. Mucor , Penicillium spp. , Fusarium spp. , Rhizopus , Alternaria and Cladosporium spp. . The isolated Aspergillus strains were A. flavus , A. fumigatus and A. niger Conclusions and potential relevance: It could be concluded that this research established the significance of the dietary addition of ginger, garlic and their combination as antifungal agents in the quail meal whereas, the garlic was the superior supplement for an obvious and considerable decreasing of yeast and mold loads in quails meat.

Antimicrobial activity of manogepix, a first-in-class antifungal, and comparator agents tested against contemporary invasive fungal isolates from an international surveillance programme (2018–2019)

Manogepix, the active moiety of the prodrug fosmanogepix, is a novel antifungal with activity against major fungal pathogens including Candida (except Candida krusei), Aspergillus and difficult-to-treat/rare moulds. We tested manogepix and comparators against 2669 contemporary (2018-2019) fungal isolates collected from 82 medical centres in North America (42.3%), Europe (37.9%), Asia-Pacific (12.3%) and Latin America (7.6%). Of these, 70.7% were Candida spp., 3.6% were non-Candida yeasts including 49 Cryptococcus neoformans var. grubii, 21.7% were Aspergillus spp. and 4.1% were other moulds. Isolates were tested for antifungal susceptibility by the CLSI reference broth microdilution method. Manogepix (MIC50/90, 0.008/0.06 mg/L) was the most active agent tested against Candida spp. isolates; corresponding anidulafungin, micafungin and fluconazole MIC90 values were 16- to 64-fold higher. Similarly, manogepix (MIC50/90, 0.5/2 mg/L) was ≥4-fold more active than anidulafungin, micafungin and fluconazole against C. neoformans var. grubii. Against Aspergillus spp., manogepix (MEC50/90, 0.015/0.03 mg/L) had comparable activity to anidulafungin and micafungin. Low manogepix concentrations inhibited uncommon species of Candida, non-Candida yeasts, and rare moulds including Scedosporium spp. and Lomentospora (Scedosporium) prolificans. Manogepix exhibited potent activity against contemporary fungal isolates, including echinocandin- and azole-resistant strains of Candida and Aspergillus spp., respectively. Although rare, Candida strains that were non-wild type for manogepix demonstrated resistance to fluconazole. However, the clinical relevance of this finding is unknown. The extended spectrum of manogepix is noteworthy for its activity against many less-common yet antifungal-resistant strains. Clinical studies are underway to evaluate the utility of fosmanogepix against difficult-to-treat resistant fungal infections.

Induction of anti-Zearalenone immune response with mimotopes identified from a phage display peptide library

Zearalenone (ZEN), a type of non-steroidal estrogenic mycotoxin, is mainly produced by several species of Fusarium molds. It is ubiquitous in contaminated grains and grain products all over the world, posing a serious threat to animal and human health. This study aims to screen the mimotopes of ZEN from a phage display random 12-mer peptide library and explore their immunogenicity. A monoclonal antibody (mAb) against ZEN was employed as the target for mimotope selection from a phage display random peptide library. After four rounds of panning, six mimotopes that could specifically bind to ZEN mAb were obtained. In order to explore the immunogenicity of these mimotopes, Balb/c mice were immunized with phages Z8, Z21, Z35, Z8:Z21:Z35(1:1:1) and the conjugate of ZEN-bovine serum albumin (ZEN-BSA), respectively. The titers of antibodies in the mice immunized with mimotopes were 1:3200 (Z8), 1:3200 (Z21), 1:6400 (Z35), 1:6400 (1:1:1 mixture of Z8, Z21 and Z35), and the binding between serum antibodies and ZEN-OVA could be blocked by ZEN standards. These results demonstrated that the mimotopes of ZEN could induce specific antibodies against ZEN, suggesting that these displayed peptides were immunogenic.

In Vitro Activity of Novel Antifungal Olorofim against Filamentous Fungi and Comparison to Eight Other Antifungal Agents.

Olorofim is a novel antifungal drug that belongs to the orotomide drug class which inhibits fungal dihydroorotate dehydrogenase (DHODH), thus halting pyrimidine biosynthesis and ultimately DNA synthesis, cell growth and division. It is being developed at a time when many invasive fungal infections exhibit antifungal resistance or have limited treatment options. The goal of this study was to evaluate the in vitro effectiveness of olorofim against a large collection of recently isolated, clinically relevant American mold isolates. In vitro antifungal activity was determined for 246 azole-susceptible Aspergillus fumigatus isolates, five A. fumigatus with TR34/L98H-mediated resistance, 19 Rhizopus species isolates, 21 Fusarium species isolates, and one isolate each of six other species of molds. Olorofim minimum inhibitory concentrations (MICs) were compared to antifungal susceptibility testing profiles for amphotericin B, anidulafungin, caspofungin, isavuconazole, itraconazole, micafungin, posaconazole, and voriconazole. Olorofim MICs were significantly lower than those of the echinocandin and azole drug classes and amphotericin B. A. fumigatus wild type and resistant isolates shared the same MIC50 = 0.008 μg/mL. In non-Aspergillus susceptible isolates (MIC ≤ 2 μg/mL), the geometric mean (GM) MIC to olorofim was 0.54 μg/mL with a range of 0.015–2 μg/mL. Olorofim had no antifungal activity (MIC ≥ 2 μg/mL) against 10% of the collection (31 in 297), including some isolates from Rhizopus spp. and Fusarium spp. Olorofim showed promising activity against A. fumigatus and other molds regardless of acquired azole resistance.