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Frontiers in Cellular and Infection... 2023Mucoromycosis is a highly aggressive angio-invasive disease of humans caused by fungi in the zygomycete order, Mucorales. While is the principal agent of mucoromycosis,...
Mucoromycosis is a highly aggressive angio-invasive disease of humans caused by fungi in the zygomycete order, Mucorales. While is the principal agent of mucoromycosis, other Mucorales fungi including , , , , and are able to cause life-threatening rhino-orbital-cerebral, pulmonary, gastro-intestinal and necrotising cutaneous infections in humans. Diagnosis of the disease currently relies on non-specific CT, lengthy and insensitive culture from invasive biopsy, and time-consuming histopathology of tissue samples. At present, there are no rapid antigen tests that detect Mucorales-specific biomarkers of infection, and which allow point-of-care diagnosis of mucoromycosis. Here, we report the development of an IgG2b monoclonal antibody (mAb), TG11, which binds to extracellular polysaccharide (EPS) antigens of between 20 kDa and 250 kDa secreted during hyphal growth of Mucorales fungi. The mAb is Mucorales-specific and does not cross-react with other yeasts and molds of clinical importance including , , , , and species. Using the mAb, we have developed a Competitive lateral-flow device that allows rapid (30 min) detection of the EPS biomarker in human serum and bronchoalveolar lavage (BAL), with a limit of detection (LOD) in human serum of ~100 ng/mL serum (~224.7 pmol/L serum). The LFD therefore provides a potential novel opportunity for detection of mucoromycosis caused by different Mucorales species.
Topics: Humans; Mucorales; Antibodies, Monoclonal; Aspergillus; Fusarium; Biomarkers
PubMed: 38145040
DOI: 10.3389/fcimb.2023.1305662 -
Biotech (Basel (Switzerland)) Dec 2023Ionic additives affect the structure, activity and stability of lipases, which allow for solving common application challenges, such as preventing the formation of...
Ionic additives affect the structure, activity and stability of lipases, which allow for solving common application challenges, such as preventing the formation of protein aggregates or strengthening enzyme-support binding, preventing their desorption in organic media. This work aimed to design a biocatalyst, based on lipase improved by the addition of ionic additives, applicable in the production of ethyl esters of fatty acids (EE). Industrial enzymes from (TLL), (RML), (CALB) and Lecitase, immobilized in commercial supports like Lewatit, Purolite and Q-Sepharose, were tested. The best combination was achieved by immobilizing lipase TLL onto Q-Sepharose as it surpassed, in terms of %EE (70.1%), the commercial biocatalyst Novozyme 435 (52.7%) and was similar to that of Lipozyme TL IM (71.3%). Hence, the impact of ionic additives like polymers and surfactants on both free and immobilized TLL on Q-Sepharose was assessed. It was observed that, when immobilized, in the presence of sodium dodecyl sulfate (SDS), the TLL derivative exhibited a significantly higher activity, with a 93-fold increase (1.02 IU), compared to the free enzyme under identical conditions (0.011 IU). In fatty acids ethyl esters synthesis, Q-SDS-TLL novel derivatives achieved results similar to commercial biocatalysts using up to ~82 times less enzyme (1 mg/g). This creates an opportunity to develop biocatalysts with reduced enzyme consumption, a factor often associated with higher production costs. Such advancements would ease their integration into the biodiesel industry, fostering a greener production approach compared to conventional methods.
PubMed: 38131679
DOI: 10.3390/biotech12040067 -
Frontiers in Microbiology 2023Mucormycosis, an invasive fungal disease with severe consequences, poses a significant threat to immunocompromised individuals. However, the timely and accurate...
Mucormycosis, an invasive fungal disease with severe consequences, poses a significant threat to immunocompromised individuals. However, the timely and accurate identification of Mucorales infection continues to present difficulties. In this study, novel detection techniques utilizing recombinase polymerase amplification (RPA) and quantitative real-time polymerase chain reaction (qPCR) were developed, specifically targeting the mitochondrial gene, in order to address this challenge. The specificity of the RPA and qPCR assay was assessed by adding genomic DNAs extracted from 14 non-targeted strains, as well as human and mouse blood. No false-positive results were observed. Additionally, genomic DNAs from 13 species in five genera of order Mucorales were tested and yielded positive results in both methods. To further evaluate the sensitivity of the assays, DNAs from and were utilized, with concentrations ranging from 1 ng/μL to 1 fg/μL. The limit of detection (LoD) for the RPA assay was determined to be 1 pg., with the exception of which had a LoD of 1 ng. The LoD for the qPCR assay varied between 10 fg and 1 pg., depending on the specific species being tested. Sensitivity analysis conducted on simulated clinical samples revealed that the LoD for RPA and qPCR assays were capable of detecting DNA extracted from 10 and 10 colony forming units (CFU) conidia in 200 μL of blood and serum, respectively. Consequently, the real-time RPA and qPCR assays developed in this study exhibited favorable sensitivity and specificity for the diagnosis of mucormycosis.
PubMed: 37954252
DOI: 10.3389/fmicb.2023.1273073 -
Gastroenterology Research Oct 2023Mucormycosis is a devastating fungal infection that is usually seen in immunocompromised hosts. It is caused by fungi of the subphylum , order , with most cases caused... (Review)
Review
Mucormycosis is a devastating fungal infection that is usually seen in immunocompromised hosts. It is caused by fungi of the subphylum , order , with most cases caused by , , or species. It can involve any organ system and can disseminate in severe cases. Lately, there has been an increased number of reports for mucormycosis infection in immunocompetent patients. Gastrointestinal system involvement is rare compared to other organ systems but has been increasingly reported in the literature. Mucormycosis can affect any part of the gastrointestinal tract and lead to different presentations depending on the area of involvement. Due to the paucity of the condition, there has been no specific guidelines on how to treat gastrointestinal mucormycosis. In this review, we discuss the risk factors of gastrointestinal mucormycosis, clinical presentation, approach to diagnosis, and most recent treatment modalities for gastrointestinal mucormycosis.
PubMed: 37937225
DOI: 10.14740/gr1662 -
Poultry Science Jan 2024Fungi are pathogens that infect all types of poultry and farmers, leading to economic losses in poultry production. Fungi can be isolated from environmental samples and...
Fungi are pathogens that infect all types of poultry and farmers, leading to economic losses in poultry production. Fungi can be isolated from environmental samples and are ubiquitous in the air. This study aimed to evaluate fungal contamination in domestic duck farm environments and analyze biosecurity risk factors associated with fungal infection incidence to assess the vulnerability of the farms to fungal infection. The average fungal concentration was 203 colony-forming units (CFU)/m in the air and 365 × 10 CFU/m in the wall surface samples. Sixteen fungal genera were recovered from air and wall surface samples from 19 duck-breeding farms, Aspergillus being the most frequently isolated (air: 43.2%; wall surface: 40%). Eleven additional fungal genera (Acrophialophora, Byssochlamys, Fusarium, Lichtheimia, Paecilomyces, Penicillium, Polycephalomyces, Rhizomucor, Scopulariopsis, Talaromyces, and Thermoascus) were isolated from air samples. Also, 8 additional fungal genera (Chaetomium, Lichtheimia, Penicillium, Petriella, Rhizomucor, Rhizopus, Talaromyces, and Trichosporon) were isolated from wall surface samples. The characteristics of the poultry farms (geographic region, stocking density, breeding house type, affiliate, duck age, and season) and fungal concentrations in the air and wall surface samples were analyzed to evaluate the biosecurity risk of the farms. Fungal infections were significantly affected by high stocking density (>2 ducks/m), duck age (18-25 wk and >60 wk), and high fungal concentration in the wall surface samples (>300 × 10 CFU/m).
Topics: Animals; Ducks; Farms; Biosecurity; Chickens; Plant Breeding; Poultry; Mycoses; Risk Factors
PubMed: 37925771
DOI: 10.1016/j.psj.2023.103197 -
International Journal of Biological... Jan 2024Various nanoflowers are synthesized for enzyme immobilization. In order to increase the activity of nanoflowers, in this study, 3D flower-like structured...
Various nanoflowers are synthesized for enzyme immobilization. In order to increase the activity of nanoflowers, in this study, 3D flower-like structured organic-inorganic hybrid nanoflowers (hNFs) with various lipases Rhizomucor miehei lipase (RML), Candida antarctica lipase B (CALB), Humicola insolens lipase (HIL), Thermomyces lanuginosus lipase (TLL), Eversa® Transform 2.0 (ET) a genetically modified enzyme derived of TLL and graphene quantum dots (GQDs) were prepared and characterized.Lipase hNFs [lipase-(Cu/Co)(PO)] and lipase@GQDs hNFs [lipase@GQDs-(Cu/Co)(PO)] were straightforwardly prepared through mixing with metal ion (Cuor Co) aqueous solutions with or without GQDs. The ET@GQDs-(Cu)(PO) hNFs demonstrated 687 % higher activity than ET-(Cu)(PO) hNFs and 650 % higher activity than the free ET. Similar results were also observed with other lipase hybrid nanoflowers. For example, TLL@GQDs-(Cu)3(PO4)2 hNFs exhibited a 557 % higher activity than TLL-(Cu)(PO) hNFs and a 463 % higher activity than free TLL. Additionally, TLL@GQDs-(Co)(PO) hNFs showed a 141 % higher activity than TLL-(Co)(PO) hNFs and a 304 % higher activity than free TLL. Upon examining pH and thermal stability, it was revealed that lipase@GQDs hNFs exhibited higher activity compared to free lipase and other hNFs without GQDs. The effect of metal ions, enzyme concentrations and amount of GQDs on the morphology and enzyme activity of the lipase-hNFs was examined.
Topics: Graphite; Quantum Dots; Lipase; Enzymes, Immobilized; Hydrophobic and Hydrophilic Interactions
PubMed: 37918587
DOI: 10.1016/j.ijbiomac.2023.127817 -
3 Biotech Nov 2023This study deals with lipase immobilization on micro- and mesoporous silica-based materials. The effects of the type of support (silica MCM-41, zeolite HZSM-5 (SAR 25),...
This study deals with lipase immobilization on micro- and mesoporous silica-based materials. The effects of the type of support (silica MCM-41, zeolite HZSM-5 (SAR 25), zeolite HZSM-5 (SAR 280), and the silica-aluminas Siral 10, Siral 20, and Siral 40) were investigated on the immobilization of lipase B from (CALB) and lipase from (RML). The supports that allowed the highest immobilization efficiencies for the CALB were Siral 40 (91.4%), HZSM-5 (SAR 280) (90.6%), and MCM-41 (89.4%). Siral 20 allowed the highest immobilization efficiency for RML (97.6%), followed by HZSM-5 (SAR 25) (77.1%) and HZSM-5 (SAR 280) (62.7%). The effect of protein concentration on lipase immobilization was investigated, and the results adjusted well on the Langmuir isotherm model (R > 0.9). The maximum protein adsorption capacity of the support determined by the Langmuir model was equal to 10.64 and 20.97 mg g for CALB and RML, respectively. The effects of pH (pH 7.0 and pH 11.0) and phosphate buffer solution concentration (5 and 100 mmol L) were also investigated on lipase immobilization. The immobilization efficiency for both lipases was similar for the different pH values. The use of 100 mmol L phosphate buffer decreased the lipase immobilization efficiency. The biocatalysts (CALB-Siral 40 and RML-Siral 20) were tested in the ethyl oleate synthesis. The conversion of 61.7% was obtained at 60 °C in the reaction catalyzed by CALB-Siral 40. Both heterogeneous biocatalysts showed increased thermal stability compared with their free form. Finally, the reuse of the biocatalysts was studied. CALB-Siral 40 and RML-Siral 20 maintained about 30% of the initial conversion after 3 batches of ethyl oleate synthesis. Silica-aluminas (Siral 20 and 40) proved to be a support that allowed a high efficiency of immobilization of lipases and activity for esterification reaction.
PubMed: 37900269
DOI: 10.1007/s13205-023-03801-x -
Current Medical Mycology Mar 2023Mucormycosis (previously called zygomycosis) is a diverse group of increasingly recognized and frequently fatal mycotic diseases caused by members of the class... (Review)
Review
Mucormycosis (previously called zygomycosis) is a diverse group of increasingly recognized and frequently fatal mycotic diseases caused by members of the class zygomycetes. Mucormycosis is around 80 times more common in India, compared to other developed countries, with a frequency of 0.14 cases per 1,000 population. The most frequent causative agent of mucormycosis is the following genera from the Order , , , , , , and . The major risk factors for the development of mucormycosis are diabetic ketoacidosis, deferoxamine treatment, cancer, solid organ or bone marrow transplantations, prolonged steroid use, extreme malnutrition, and neutropenia. The common clinical forms of mucormycosis are rhino-orbital-cerebral, pulmonary, cutaneous, and gastrointestinal. During the second wave of COVID-19, there was a rapid increase in mucormycosis with more severity than before. Amphotericin B is currently found to be an effective drug as it is found to have a broad-spectrum activity and posaconazole is used as a salvage therapy. Newer triazole isavuconazole is also found effective against mucormycosis. This study aimed to review various studies on the laboratory diagnosis and treatment of mucormycosis.
PubMed: 37867589
DOI: 10.18502/cmm.2023.345032.1400 -
Foods (Basel, Switzerland) Sep 2023Shaoxing rice wine is a notable exemplar of Chinese rice wine. Its superior quality is strongly correlated with the indigenous natural environment. The results indicated...
Shaoxing rice wine is a notable exemplar of Chinese rice wine. Its superior quality is strongly correlated with the indigenous natural environment. The results indicated that Firmicutes (75%), Actinobacteria (15%), Proteobacteria (5%), and Bacteroidetes (3%) comprised the prevailing bacterial groups. Among the main bacterial genera, was the most abundant, accounting for 49.4%, followed by (11.9%), (13.1%), (4.1%), and (1.1%). The dominant fungal phyla were Ascomycota and Zygomycota. Among the dominant genera, (59.3%) prevailed as the most abundant, followed by (10.7%), (7.1%), (6.2%), (4.9%), (2.2%), and (1.3%). The findings demonstrate that the structure of the bacterial and fungal communities remains stable in the environment, with their diversity strongly influenced by climatic conditions. The continuous fluctuations in environmental factors, such as temperature, air pressure, humidity, rainfall, and light, significantly impact the composition and diversity of microbial populations, particularly the dominant bacterial community.
PubMed: 37835217
DOI: 10.3390/foods12193564 -
Cureus Sep 2023Mucormycosis is a rare opportunistic fungal infection caused by and associated with high mortality rates. Rhino-orbito-cerebral localization usually occurs in...
Mucormycosis is a rare opportunistic fungal infection caused by and associated with high mortality rates. Rhino-orbito-cerebral localization usually occurs in individuals with uncontrolled diabetes mellitus. We report the case of a 41-year-old male, with previously undiagnosed diabetes, who presented with unilateral facial extensive black eschar and signs of diabetic ketoacidosis. Cerebral magnetic resonance imaging showed left pansinusitis, left craniofacial edematous infiltrate, and left proptosis. A left internal temporal abscess was identified at an early pre-suppurative stage. Magnetic resonance angiography revealed total occlusion of the left intracranial internal carotid artery. A histopathological study of nasal mucosa biopsy suggested mucormycosis. According to the clinical presentation and the radiological and histopathological findings, rhino-orbito-cerebral mucormycosis was presumed. Culture of nasal, ocular, and skin lesion specimens grew . and confirmed the diagnosis. The patient was treated with systemic liposomal amphotericin B. He died of multiple organ failure before surgical debridement was possible as he was in critical condition requiring stabilization before surgical treatment.
PubMed: 37809164
DOI: 10.7759/cureus.44768