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Despite the severe impact of uncommon yeast fungal infections and the pressing need for more research on the topic, there are still few studies available on the identification, epidemiology, and susceptibility profile of those pathogens. The aims of the current study were to define the profile of uncommon yeast species at Fattouma Bourguiba University Hospital using phenotypic, molecular, and proteomic methods and to study their antifungal susceptibility profile. Pre-identified uncommon yeast species were collected from 2018 to 2021. These isolates were further identified using phenotypic methods (ID32C® system and Vitek2® YST), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and sequencing. The antifungal susceptibility profile was studied using the reference CLSI broth microdilution method. In total, 30 strains were collected during the study period. Referring to the sequencing, the most isolated uncommon species were Saprochaete capitata, Candida lusitaniae, Candida kefyr, Candida inconspicua, and Candida guilliermondii. A total of 90% of isolates were correctly identified by MALDI-TOF MS compared to 76.7% and 63.3% by ID32® C and VITEK® 2 YST, respectively. The isolated species showed variable responses to antifungals. Candida guilliermondii showed increased azole minimum inhibitory concentrations. Misidentification of uncommon yeast species was common using commercial phenotypic methods. The high percentage of concordance of MALDI-TOF results with sequencing highlights its high performance and usefulness as a routine diagnosis tool., (© The Author(s) 2024. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Zoonotic yeast species have been implicated in disease development in both humans and cats. This study analyzed the yeast mycobiota present in feline facial hair and human nails and explored potential interspecies associations. A total of 118 biological specimens were examined, including 59 feline facial hair and 59 human nail samples. DNA extraction and DNA sequencing were performed to identify the specific yeast species. The most predominant yeast species in humans and cats were selected for antifungal susceptibility testing (itraconazole, ketoconazole, miconazole, and terbinafine). The findings unveiled diverse yeast species in cats and humans. Malassezia pachydermatis (45.8%) and Malassezia furfur (30.5%) were the most common yeast species in cats and humans, respectively. However, no significant correlation was detected between the yeast species identified in cats and their owners residing in the same household (p > 0.05). Miconazole exhibited the highest minimum inhibitory concentrations (MICs) against Malassezia pachydermatis and Malassezia furfur in both cat and human isolates, whereas terbinafine showed the lowest MICs against most Malassezia pachydermatis and Malassezia furfur in both cat and human isolates. Diverse yeast species in cat facial hair and human nails suggest possible cross-contamination among humans, pets, and environments., (© 2024. The Author(s).)

Yeast complexes in the fecal samples of wild (Dendrocopos major, Picus viridis) and partially synanthropic (Bombycilla garrulus, Garrulus glandarius, Pica pica, and Pyrrhula pyrrhula) birds were studied in a forest ecosystem during winter. A total of 18 yeast species were identified: 16 ascomycetes and two basidiomycetes belonging to five subphyla of fungi: Saccharomycotina (15), Pezizomycotina (1), Agaricomycotina (1), and Pucciniomycotina (1). Most yeast species were found in the fecal samples of P. pyrrhula (Candida parapsilosis, C. zeylanoides, Debaryomyces hansenii, Hanseniaspora uvarum, Metschnikowia pulcherrima, Meyerozyma carpophila, M. guilliermondii, Rhodotorula mucilaginosa); the lowest number of yeast species was observed in the feces of B. garrulus (C. parapsilosis, C. zeylanoides, Met. pulcherrima, and Rh. mucilaginosa). The opportunistic species of the genus Candida were found only in feces of partially synanthropic birds: C. parapsilosis was observed in the feces of B. garrulus, G. glandarius, P. pica, and P. pyrrhula; its relative abundance was 69.3%, 49.1%, 10.5%, and 1.1%, respectively; C. tropicalis was observed in the feces of P. pica and G. glandarius; its relative abundance was 54.6% and 7.1%, respectively. Strains of C. parapsilosis and C. tropicalis isolated from the feces of partially synanthropic birds were evaluated for their susceptibility to conventional antifungal agents (fluconazole, voriconazole, amphotericin B) and hydrolytic activity. A total of 160 strains were studied. Resistance to fluconazole was detected in 86.8% of C. parapsilosis strains and in 87% of C. tropicalis strains; resistance to voriconazole was detected in 71.7% of C. parapsilosis and in 66.7% of C. tropicalis strains, and the lowest percentage of resistant strains was detected to amphotericin B, 2.8% and 3.7% in C. parapsilosis and C. tropicalis strains, respectively. Multiresistance was detected in one strain of C. parapsilosis isolated from P. pica feces and in one strain of C. tropicalis isolated from G. glandarius feces. Phospholipase and hemolysin activities in the strains of C. parapsilosis were low (mean Pz values of 0.93 and 0.91, respectively); protease activity was moderate (mean Pz value of 0.53). The ability to produce hydrolytic enzymes was higher in the isolated strains of C. tropicalis. The mean Pz values of phospholipase and hemolysin activities were moderate (mean Pz values of 0.63 and 0.60, respectively), whereas protease activity was high (mean Pz value of 0.32). Thus, wild and partially synanthropic birds play an important role in disseminating of various yeast species. These yeasts can enter the topsoil via feces and contribute to the formation of allochthonous and uneven soil yeast diversity in natural ecosystems. In addition, partially synanthropic birds can be vectors of virulent strains of opportunistic Candida species from urban environments to natural biotopes., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Although Candida species are the most common cause of fungemia, non-Candida rare yeasts (NCY) have been increasingly reported worldwide. Although the importance of these yeast infections is recognized, current epidemiological information about these pathogens is limited, and they have variable antifungal susceptibility profiles. In this study, we aimed to evaluate the clinical characteristics for fungemia caused by NCY by comparing with candidemia. The episodes of NCY fungemia between January 2011 and August 2023 were retrospectively evaluated in terms of clinical characteristics, predisposing factor, and outcome. In addition, a candidemia group, including patients in the same period was conducted for comparison. Antifungal susceptibility tests were performed according to the reference method. A total of 85 patients with fungemia episodes were included: 25 with NCY fungemia and 60 with candidemia. Fluconazole had high minimal inhibitory concentration (MIC) values against almost all NCY isolates. The MIC values for voriconazole, posaconazole, and amphotericin B were ≤ 2 µg/ml, and for caspofungin and anidulafungin were ≥ 1 µg/ml against most of isolates. Hematological malignancies, immunosuppressive therapy, neutropenia and prolonged neutropenia, polymicrobial bacteremia/fungemia, preexposure to antifungal drugs, and breakthrough fungemia were associated with NCY fungemia, whereas intensive care unit admission, diabetes mellitus, urinary catheters, and total parenteral nutrition were associated with candidemia. In conclusion, the majority of fungemia due to NCY species was the problem, particularly in hematology units and patients with hematological malignancy. Preexposure to antifungal drugs likely causes a change in the epidemiology of fungemia in favor of non-albicans Candida and/or NCY., (© The Author(s) 2024. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

This research paper presents a novel approach to the green synthesis of silver nanoparticles (AgNPs) using viticultural waste, allowing to obtain NP dispersions with distinct properties and morphologies (monodisperse and polydisperse AgNPs, referred to as mAgNPs and pAgNPs) and to compare their biological activities. Our synthesis method utilized the ethanolic extract of Vitis vinifera pruning residues, resulting in the production of mAgNPs and pAgNPs with average sizes of 12 ± 5 nm and 19 ± 14 nm, respectively. Both these AgNPs preparations demonstrated an exceptional stability in terms of size distribution, which was maintained for one year. Antimicrobial testing revealed that both types of AgNPs inhibited either the growth of planktonic cells or the metabolic activity of biofilm sessile cells in Gram-negative bacteria and yeasts. No comparable activity was found towards Gram-positives. Overall, pAgNPs exhibited a higher antimicrobial efficacy compared to their monodisperse counterparts, suggesting that their size and shape may provide a broader spectrum of interactions with target cells. Both AgNP preparations showed no cytotoxicity towards a human keratinocyte cell line. Furthermore, in vivo tests using a silkworm animal model indicated the biocompatibility of the phytosynthesized AgNPs, as they had no adverse effects on insect larvae viability. These findings emphasize the potential of targeted AgNPs synthesized from viticultural waste as environmentally friendly antimicrobial agents with minimal impact on higher organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objective: The aim was to systematically review clinical studies that investigated the efficacy of antimicrobial photodynamic therapy (aPDT) in reducing oral yeasts growth (OYG) in individuals wearing implant overdentures (IO)., Methods: The focused question was "Is aPDT effective in reducing OYG in patients wearing IO?" Literature search was performed in accordance with PRISMA guidelines. Indexed databases were searched without time and language restrictions up to and including January 2024. Clinical studies were included; and letters to the Editor, case-reports/case-series, perspectives/commentaries, in-vitro/ex-vivo studies, studies on animal models and expert opinions were excluded. The risk of bias was also assessed., Results: Two clinical studies were included and processed for data extraction. The study population comprised of 100 (mean age: 58.5 years) and 53 (mean age: 58.5 years) individuals. The numbers of males and females included in these studies ranged between 33 and 35 males and 18-67 females, respectively. In both studies, follow-up evaluations were performed after 60 days. In both studies, aPDT was performed using a 660 nm diode laser at a power of 100 mW and using methylene-blue as photosensitizer. Results from both studies showed that aPDT is effective in significantly reducing oral yeasts CFU/ml and improvement of OHRQoL of individuals using IO., Conclusion: The aPDT is useful in reducing OYG on IO; however, further well-designed and power-adjusted studies are needed in this area of research., Competing Interests: Declaration of competing interest The author declares no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Formaldehyde and acetaldehyde are reactive small molecules produced endogenously in cells as well as being environmental contaminants. Both of these small aldehydes are classified as human carcinogens, since they are known to damage DNA and exposure is linked to cancer incidence. However, the mutagenic properties of formaldehyde and acetaldehyde remain incompletely understood, at least in part because they are relatively weak mutagens. Here, we use a highly sensitive yeast genetic reporter system featuring controlled generation of long single-stranded DNA regions to show that both small aldehydes induced mutational patterns characterized by predominantly C/G → A/T, C/G → T/A, and T/A → C/G substitutions, each in similar proportions. We observed an excess of C/G → A/T transversions when compared to mock-treated controls. Many of these C/G → A/T transversions occurred at TC/GA motifs. Interestingly, the formaldehyde mutational pattern resembles single base substitution signature 40 from the Catalog of Somatic Mutations in Cancer. Single base substitution signature 40 is a mutational signature of unknown etiology. We also noted that acetaldehyde treatment caused an excess of deletion events longer than 4 bases while formaldehyde did not. This latter result could be another distinguishing feature between the mutational patterns of these simple aldehydes. These findings shed new light on the characteristics of 2 important, commonly occurring mutagens., (© The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America.)

Heavy metals act as cofactors for several microbial enzymes and are required in low concentrations for the proper biological functioning of yeasts. Because concentrations beyond the permitted threshold can damage a cell's functionality and viability, metal tolerance in yeasts towards such heavy metals is therefore desirable during fermentation. Tyrosol, a quorum-sensing molecule in yeasts, protects yeasts from oxidative stress induced by various factors, but the performance of the molecule under heavy metal-induced stress is not known. In this investigation, the metal tolerance of four species of endemic yeasts from northeast India, Wickerhamomyces anomalus, Candida tropicalis, Saccharomyces cerevisiae and Candida glabrata, isolated from traditional starter culture cakes, was tested towards zinc (Zn+2), manganese (Mn+2), cobalt (Co+2) and copper (Cu+2) in the presence and absence of tyrosols retrieved from these isolates. The decreasing order of the tolerance of isolates was found to be Mn+2 > Zn+2 > Co+2 > Cu+2. Under the influence of tyrosols, isolates showed enhanced growth at their upper metal tolerance limit. Candida tropicalis showed enhanced growth (2-48-fold, P < 0.0001) in all the tested metal consisting medium (2 mM Zn+2, 5 mM Mn+2, 2 mM Co+2 and 1 mM Cu+2), while W. anomalus, C. glabrata and S. cerevisiae showed increased growth (3-17-fold, P < 0.0001) in Zn+2 (2 mM), Mn+2 (5 mM) and Cu+2 (1 mM) augmented medium. The overall result suggests that tyrosol exerts a protective effect under heavy metal-induced stress, which could be useful in enhancing the quality of fermented products., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS.)

4,5,6,7-Tetrabromo-1H-benzotriazole is widely used as the reference ATP-competitive inhibitor of protein kinase CK2. Herein, we study its new analogs: 5,6-diiodo- and 5,6-diiodo-4,7-dibromo-1H-benzotriazole. We used biophysical (MST, ITC) and biochemical (enzymatic assay) methods to describe the interactions of halogenated benzotriazoles with the catalytic subunit of human protein kinase CK2 (hCK2α). To trace the biological activity, we measured their cytotoxicity against four reference cancer cell lines and the effect on the mitochondrial inner membrane potential. The results obtained lead to the conclusion that iodinated compounds are an attractive alternative to brominated ones. One of them retains the cytotoxicity against selected cancer cell lines of the reference TBBt with a smaller side effect on mitochondrial activity. Both iodinated compounds are candidate leaders in the further development of CK2 inhibitors., (© 2021. The Author(s).)

The essential oils of three specimens of Myrcia multiflora (A, B and C) and Eugenia florida were extracted by hydrodistillation, and the chemical compositions from the essential oils were identified by gas chromatography and flame ionization detection (CG/MS and CG-FID). The fungicide potential of the EOs against five fungicide yeasts was assessed: Candida albicans INCQS-40175, C. tropicalis ATCC 6258, C. famata ATCC 62894, C. krusei ATCC 13803 and C. auris IEC-01. The essential oil of the specimen Myrcia multiflora (A) was characterized by the major compounds: α-bulnesene (26.79%), pogostol (21.27%) and δ-amorphene (6.76%). The essential oil of the specimen M. multiflora (B) was rich in ( E )-nerolidol (44.4%), (E)-γ-bisabolene (10.64%) and ( E , E )-α-farnesene (8.19%), while ( E )-nerolidol (92.21%) was the majority of the specimen M. multiflora (C). The sesquiterpenes seline-3,11-dien-6-α-ol (12.93%), eremoligenol (11%) and γ-elemene (10.70%) characterized the chemical profile of the EOs of E. florida . The fungal species were sensitive to the essential oil of M. multiflora (B) (9-11 mm), and the lowest inhibitory concentration (0.07%) was observed in the essential oil of M. multiflora (A) against the yeasts of C. famata . Fungicidal action was observed in the essential oils of M. multiflora (A) against C. famata , with an MIC of 0.78 µL/mL and 3.12 µL/mL; C. albicans , with an MFC of 50 µL/mL and M. multiflora (C) against C. albicans ; and C. krusei , with a MFC of 50 µL/mL.

One of the greatest threats to human and animal health is posed by infections caused by drug-resistant bacterial strains. Therefore, newly synthesised substances are tested for their antimicrobial activity. Carbazole derivatives seem to be promising antibacterial agents. This study aimed at investigating the toxicity and activity of newly synthesised, functionalised carbazole derivative 2 (4-(4-(benzylamino)butoxy)-9 H -carbazole) against various microorganisms. Its antimicrobial potential against Gram-positive and Gram-negative bacteria, yeast, and filamentous fungi was examined according to CLSI (Clinical and Laboratory Standards Institute) standards. The tested compound was found to efficiently inhibit the growth of Gram-positive strains. The addition of carbazole derivative 2 at the concentration of 30 µg/mL caused inhibition of bacterial growth by over 95%. Moreover, about 50 and 45% limitation of Pseudomonas aeruginosa and Aspergillus flavus growth was observed in the samples incubated with the addition of 20 and 60 µg/mL of the compound, respectively. Its addition to the microbial cultures caused an increase in the permeability of the cellular membrane. Slight haemolysis of red blood cells was observed after 24-h treatment with carbazole derivative 2 . On the other hand, human fibroblasts were found to be more sensitive to its effects. The activity of the tested compound indicates a possibility of its further modification in order to obtain effective drugs, especially against drug-resistant staphylococci.

Silymarin is known for its hepatoprotective effects. Although there is solid evidence for its protective effects against Amanita phalloides intoxication, only inconclusive data are available for alcoholic liver damage. Since silymarin flavonolignans have metal-chelating activity, we hypothesized that silymarin may influence alcoholic liver damage by inhibiting zinc-containing alcohol dehydrogenase (ADH). Therefore, we tested the zinc-chelating activity of pure silymarin flavonolignans and their effect on yeast and equine ADH. The most active compounds were also tested on bovine glutamate dehydrogenase, an enzyme blocked by zinc ions. Of the six flavonolignans tested, only 2,3-dehydroderivatives (2,3-dehydrosilybin and 2,3-dehydrosilychristin) significantly chelated zinc ions. Their effect on yeast ADH was modest but stronger than that of the clinically used ADH inhibitor fomepizole. In contrast, fomepizole strongly blocked mammalian (equine) ADH. 2,3-Dehydrosilybin at low micromolar concentrations also partially inhibited this enzyme. These results were confirmed by in silico docking of active dehydroflavonolignans with equine ADH. Glutamate dehydrogenase activity was decreased by zinc ions in a concentration-dependent manner, and this inhibition was abolished by a standard zinc chelating agent. In contrast, 2,3-dehydroflavonolignans blocked the enzyme both in the absence and presence of zinc ions. Therefore, 2,3-dehydrosilybin might have a biologically relevant inhibitory effect on ADH and glutamate dehydrogenase.

Tailoring nanomaterials with tunable properties is of great importance to develop multifunctional candidates in the biomedical field. In the present study, we aimed to develop a promising nano-hybrid system composed of chitosan (CS) and mesoporous silica nanoparticles with a silver nanoshell coat (CS-AgMSNs). The physicochemical properties of CS-AgMSNs films were characterized using various techniques. Further, the mechanical properties of CS-AgMSNs were evaluated and compared with those of undoped CS film. Moreover, the antimicrobial activities of CS-AgMSNs (with different concentrations) were assessed against E-coli, S. aureus, C. albicans, and A. niger. Our results demonstrated that increasing the concentrations of doped AgMSNs (10 to 40 mg) in CS films lowered their transparency and blocked light transmission effectively. The measured elastic modulus of CS-AgMSNs films (20 and 30 mg) showed a decrease in the stiffness of CS films. Also, the elongation at break for CS-AgMSNs (40 mg) indicated a better flexibility. CS-AgMSNs films (10-40 mg) showed an enhanced antimicrobial activity in a concentration-dependent manner compared to undoped CS films. Collectively, the results suggest that our nano-hybrid CS-AgMSNs matrix has unique and promising properties, and holds potential for use in the biomedical field, food packaging, and textile industry., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Herein, tannic acid (TA)-reinforced chitosan (CHS)/β-cyclodextrin (β-CD) biocomposite membranes were prepared by TA solution incubating treatment. The functional groups, crystal structure, and morphological characterizations of the prepared biocomposite membranes were investigated using various methods. The biocomposite membranes were investigated in terms of their wettability, porosity, swelling degree, and water uptake. In vitro antioxidant investigation was carried out through DPPH assay. Moreover, the prepared biocomposite membranes were evaluated for their antimicrobial ability against three different microbial species. The introduction of TA effectively improved the swelling behavior, mechanical strength, and porosity of the biocomposite membranes. TA increased the tensile strength from 0.7 ± 0.2 MPa to a maximum of 2.2 ± 0.6 MPa and elongation at break from 26.9 ± 0.7% to a maximum of 36.7 ± 3.5%. The biocomposite membranes showed an initial burst release of TA (~40%) within 6 h, followed by a gradual release of 100% by 18 h. Furthermore, the introduction of TA into the biocomposite membranes further improved the antimicrobial activities against both bacteria and yeast, as well as the in vitro antioxidant potential. As a consequence, the prepared biocomposite membranes could potentially be used as scaffold in broaden biomedical fields due to their adaptable structure, porosity, greatly antioxidant, and antimicrobial activity., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The mass production of graphene oxide (GO) unavoidably elevates the chance of human exposure, as well as the possibility of release into the environment with high stability, raising public concern as to its potential toxicological risks and the implications for humans and ecosystems. Therefore, a thorough assessment of GO toxicity, including its potential reliance on key physicochemical factors, which is lacking in the literature, is of high significance and importance. In this study, GO toxicity, and its dependence on oxidation level, elemental composition, and size, were comprehensively assessed. A newly established quantitative toxicogenomic-based toxicity testing approach, combined with conventional phenotypic bioassays, were employed. The toxicogenomic assay utilized a GFP-fused yeast reporter library covering key cellular toxicity pathways. The results reveal that, indeed, the elemental composition and size do exert impacts on GO toxicity, while the oxidation level exhibits no significant effects. The UV-treated GO, with significantly higher carbon-carbon groups and carboxyl groups, showed a higher toxicity level, especially in the protein and chemical stress categories. With the decrease in size, the toxicity level of the sonicated GOs tended to increase. It is proposed that the covering and subsequent internalization of GO sheets might be the main mode of action in yeast cells.

Plant cysteine-rich peptides (CRPs) represent a diverse group of molecules involved in different aspects of plant physiology. Antimicrobial peptides, which directly suppress the growth of pathogens, are regarded as promising templates for the development of next-generation pharmaceuticals and ecologically friendly plant disease control agents. Their oligopeptide fragments are even more promising because of their low production costs. The goal of this work was to explore the antimicrobial activity of nine short peptides derived from the γ-core-containing regions of tomato CRPs against important plant and human pathogens. We discovered antimicrobial activity in peptides derived from the defensin-like peptides, snakins, and MEG, which demonstrates the direct involvement of these CRPs in defense reactions in tomato. The CRP-derived short peptides appeared particularly active against the gram-positive bacterium Clavibacter michiganensis, which causes bacterial wilt-opening up new possibilities for their use in agriculture to control this dangerous disease. Furthermore, high inhibitory potency of short oligopeptides was demonstrated against the yeast Cryptococcus neoformans , which causes serious diseases in humans, making these peptide molecules promising candidates for the development of next-generation pharmaceuticals. Studies of the mode of action of the two most active peptides indicate fungal membrane permeabilization as a mechanism of antimicrobial action.

Molecules involved in DNA damage response (DDR) are often overexpressed in cancer cells, resulting in poor responses to chemotherapy and radiotherapy. Although treatment efficacy can be improved with the concomitant use of DNA repair inhibitors, the accompanying side effects can compromise the quality of life of patients. Therefore, in this study, we identified a natural compound that could inhibit DDR, using the single-strand annealing yeast-cell analysis system, and explored its mechanisms of action and potential as a chemotherapy adjuvant in hepatocellular carcinoma (HCC) cell lines using comet assay, flow cytometry, Western blotting, immunofluorescence staining, and functional analyses. We developed a mouse model to verify the in vitro findings. We found that hydroxygenkwanin (HGK) inhibited the expression of RAD51 and progression of homologous recombination, thereby suppressing the ability of the HCC cell lines to repair DNA damage and enhancing their sensitivity to doxorubicin. HGK inhibited the phosphorylation of DNA damage checkpoint proteins, leading to apoptosis in the HCC cell lines. In the mouse xenograft model, HGK enhanced the sensitivity of liver cancer cells to doxorubicin without any physiological toxicity. Thus, HGK can inhibit DDR in liver cancer cells and mouse models, making it suitable for use as a chemotherapy adjuvant.

Chitosan (Ch) was reacted with seven benzaldehyde analogs separately through reductive amination in which the corresponding imines were formed and followed by reduction to produce N-(benzyl) chitosan (NBCh) derivatives. 1 H NMR spectroscopy was used to characterize the products. The nanoparticles (NPs) of Ch and NBCh derivatives were prepared according to the ionotropic gelation mechanism between Ch products and sodium tripolyphosphate, followed by high-energy ultrasonication. Scanning electron microscopy, particle size, polydispersity index, and zeta potential were applied for the NPs examination. The particle size was ranged from 235.17 to 686.90 nm and narrow size distribution (PDI <1). The zeta potential of NPs was varied between -1.26 and -27.50 mV. The antimicrobial activity was evaluated against bacteria (Erwinia carotovora subsp. atroseptica, Erwinia carotovora subsp. carotovora, and Ralstonia solanacearum), fungi (Aspergillus flavus and Aspergillus niger), and yeast (Candida albicans). The action of NBCh derivatives was significantly higher than Ch. The NPs had considerably higher than the Ch and NBCh derivatives. The activity was directly proportional to the chemical derivatization of Ch and the zeta potential of the NPs. The antimicrobial efficacy of these derivatives formulated in a greener approach could become an alternative to using traditional antimicrobial applications in an environmentally friendly manner., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Lifestyle factors such as hair length, the frequency of ear cleaning and bathing, age, cat rearing, and sex may contribute to opportunistic yeast infections in the external ear canal of cats. This study aimed to determine the prevalence of commensal yeast organisms in cats' external ear canals, evaluate their predisposing lifestyle factors, and test the susceptibility of Malassezia pachydermatis to antifungal agents., Results: A total of 53 cats (33 male and 20 female) seronegative for feline leukemia virus and feline immunodeficiency virus were enrolled in this study. Their mean age (± standard deviation) was 6.04 (± 3.49) years. Fungal cultures and polymerase chain reaction tests were performed to identify the yeast species derived from the external ear canal. The association between lifestyle factors and the presence of M. pachydermatis was evaluated using Fisher's exact test. The susceptibility of M. pachydermatis to antifungal agents was also analyzed. M. pachydermatis was the most frequently recovered yeast species, with a prevalence of 50.94 % (95 % confidence interval [CI]: 36.84-64.94 %). There was an association between hair length and a positive culture for M. pachydermatis (p = 0.0001). The odds of a negative culture for M. pachydermatis among short-haired cats was 11.67 (95 % CI, 3.22-42.24) times higher than that among long-haired cats (p = 0.0002). There was also an association between the frequency of ear cleaning and the presence of M. pachydermatis (p = 0.007). The odds of a negative culture for M. pachydermatis in cats that were receiving ear cleaning at intervals of ≤ 2 weeks was 5.78 (95 % CI, 1.67-19.94) times greater than that of cats receiving ear cleaning at intervals greater than 2 weeks or never (p = 0.0055). Ranges of minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations for itraconazole, ketoconazole, miconazole, and terbinafine against M. pachydermatis were ≤ 0.063-4 and ≤ 0.063-≥32, ≤ 0.063-8 and 0.125-≥32, ≤ 0.063-≥32 and 0.5-≥32, and ≤ 0.016-1 and 0.125-8 µg/ml, respectively., Conclusions: M. pachydermatis was the most commonly identified yeast organism in the external ear canal of healthy cats. Hair length and the frequency of ear cleaning played a role in the colonization of M. pachydermatis. The M. pachydermatis isolates had various MIC levels for common fungicides., (© 2021. The Author(s).)

Objectives: New automated modules are required to provide fully automated solutions in diagnostic microbiology laboratories. We evaluated the performance of a Becton Dickinson Kiestra™ IdentifA/SusceptA prototype for MALDI-TOF identification (ID) and Phoenix™ antibiotic susceptibility testing (AST)., Methods: The performance of the IdentifA/SusceptA coupled prototype was compared with manual processing for MALDI-TOF ID on 1302 clinical microbial isolates or ATCC strains and for Phoenix™ M50 AST on 484 strains, representing 61 species., Results: Overall, the IdentifA exhibited similar ID performances than manual spotting. Higher performances were observed for Gram-negative bacteria with an ID at the species level (score >2) of 96.5% (369/382) and 86.9% (334/384), respectively. A significantly better performance was observed with the IdentifA (95.2%, 81/85) compared with manual spotting (75.2%, 64/85) from colonies on MacConkey agar. Contrariwise, the IdentifA exhibited lower ID performances at the species level than manual processing for streptococci (76.1%, 96/126 compared with 92%, 115/125), coagulase-negative staphylococci (73.3%, 44/60 compared with 90%, 54/60) and yeasts (41.3%, 19/46 compared with 78.2%, 36/46). Staphylococcus aureus and enterococci were similarly identified by the two approaches, with ID rates of 92% (65/70) for the IdentifA and 92.7%, (64/69) for manual processing and 94.8%, (55/58) for the IdentifA and 98.2%, (57/58) for manual processing, respectively. The SusceptA exhibited an AST overall essential agreement of 98.82% (6863/6945), a category agreement of 98.86% (6866/6945), 1.05% (6/570) very major errors, 0.16% (10/6290) major errors, and 0.91% (63/6945) minor errors compared to the reference AST., Conclusions: Overall, the automated IdentifA/SusceptA exhibited high ID and AST performances., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Onychomycoses are difficult-to-treat fungal infections with a high recurrence rate that relates to the anatomic and pathophysiological conditions in the nail organ and the required extended duration of treatment. Clinical-epidemiological studies demonstrated that non-dermatophyte molds and yeasts are the primary causative agents in 20%-30% of onychomycoses. Mixed infections with dermatophytes are observed as well. Therefore, the causative agents should be determined by fungal culture and the antifungal treatment regimen should reliably cover non-dermatophytes, if appropriate. Systemic-topical combination therapy involving a broad-spectrum, locally applied antifungal may increase the mycological and clinical cure rates compared to monotherapy with systemic drugs., (© 2021 Wiley-VCH GmbH.)

Indole-3-acetic acid (IAA) is the most common plant hormone of the auxin class and regulates various plant growth processes. The present study investigated IAA production by the basidiomycetous yeast Rhodosporidiobolus fluvialis DMKU-CP293 using the one-factor-at-a-time (OFAT) method and response surface methodology (RSM). IAA production was optimized in shake-flask culture using a cost-effective medium containing 4.5% crude glycerol, 2% CSL and 0.55% feed-grade L-tryptophan. The optimized medium resulted in a 3.3-fold improvement in IAA production and a 3.6-fold reduction in cost compared with those obtained with a non-optimized medium. Production was then scaled up to a 15-L bioreactor and to a pilot-scale (100-L) bioreactor based on the constant impeller tip speed (V tip ) strategy. By doing so, IAA was successfully produced at a concentration of 3569.32 mg/L at the pilot scale. To the best of our knowledge, this is the first report of pilot-scale IAA production by microorganisms. In addition, we evaluated the effect of crude IAA on weed growth. The results showed that weed (Cyperus rotundus L.) growth could be inhibited by 50 mg/L of crude IAA. IAA therefore has the potential to be developed as a herbicidal bioproduct to replace the chemical herbicides that have been banned in various countries, including Thailand.

Introduction: Onychomycosis are infections with a variety of etiological agents. Although dermatophytes are responsible for most infections, yeasts are gaining importance as agents of these pathologies. The use of antifungals has increased the incidence of what had been considered rare or novel pathogens. We reidentify three rare yeasts from a culture collection of onychomycosis agents by matrix-assisted laser desorption/ionization time of flight/mass spectrometry (MALDI-TOF/MS) and sequencing the internal transcribed spacer (ITS) regions or the intergenic spacer (IGS) 1 region of ribosomal DNA (rDNA), and present their enzymatic and antifungal susceptibility profiles., Material and Methods: We performed a phenotypical characterization and molecular identification of five yeast isolates. We tested the urease, gelatinase, DNase, phospholipase, protease, and esterase activities, as well as the hemolytic activity. We evaluated the antifungal susceptibility to amphotericin B, fluconazole, anidulafungin and caspofungin., Results: Phenotypic methods could not identify the isolates. MALDI-TOF/MS was able to properly identify Candida duobushameulonii. The five isolates were successfully identified by sequence analysis as Candida duobushaemulonii, Meyerozyma caribbica and Cutaneotrichosporon dermatis. Candida duobushameulonii showed hemolytic, phospholipase, and protease activities. Meyerozyma caribbica was positive for gelatinase and protease activities. All antifungals exhibited minimum inhibitory concentrations (MICs) ≤2μg/mL against both species. The three isolates of Cutaneotrichosporon dermatis showed urease, DNase, and esterase activities, and resistance to echinocandins (MICs ≥8μg/mL), while amphotericin B and fluconazole exhibited low MICs against these isolates (0.50-2μg/mL)., Discussion: Sequencing of the ITS or IGS1 regions of rDNA remains the best method for identifying cryptic species over other commercially available systems. More reports are needed to define the enzymatic and antifungal profiles for these species. This is the first report of Meyerozyma caribbica and Cutaneotrichosporon dermatis as etiological agents of onychomycosis., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Lignocellulosic hydrolysates will also contain compounds that inhibit microbial metabolism, such as organic acids, furaldehydes, and phenolic compounds. Understanding the response of yeasts toward such inhibitors is important to the development of different bioprocesses. In this work, the growth capacity of 7 industrial Saccharomyces cerevisiae and 7 non-Saccharomyces yeasts was compared in the presence of 3 different concentrations of furaldehydes (furfural and 5-hydroxymetil-furfural), organic acids (acetic and formic acids), and phenolic compounds (vanillin, syringaldehyde, ferulic, and coumaric acids). Then, Candida tropicalis JA2, Meyerozyma caribbica JA9, Wickerhamomyces anomalus 740, S. cerevisiae JP1, B1.1, and G06 were selected for fermentation in presence of acetic acid, HMF, and vanillin because they proved to be most tolerant to the tested compounds, while Spathaspora sp. JA1 because its xylose consumption rate. The results obtained showed a dose-dependent response of the yeasts toward the eight different inhibitors. Among the compared yeasts, S. cerevisiae strains presented higher tolerance than non-Saccharomyces, 3 of them with the highest tolerance among all. Regarding the non-Saccharomyces yeasts, C. tropicalis JA2 and W. anomalus 740 appeared as the most tolerant, whereas Spathaspora strains appeared very sensitive to the different compounds.

Combination effects of ɛ-polylysine coating (0.5 and 1%) and stinging nettle extract (3, 6 and 9%) on quality properties and shelf life of beef meat (2 × 2 × 2 cm) was evaluated at 4 °C for 12 days. The results indicated that ɛ-polylysine (ε-PL) coating with stinging nettle extract (SNE) had no significant effects on ash, fat, protein and moisture content among packaged beef samples in polyethylene bags (in atmosphere condition). At the end of storage, beef samples coated with 1% ε-PL and 9% SNE had significantly lower TBARS and TVB-N values compared to those found in control. Furthermore, 1% ε-PL coating with SNE 9% showed the highest inhibitory effects against molds and yeast, total viable counts (TVC) and coliforms during storage. However, sensory evaluation showed that samples coated with 1% ε-PL and 6% SNE had the highest scores for overall acceptability compared to the other groups. Based on the obtained results, ε-PL coating with SNE could be effectively used for extending the beef meat shelf life without negative effects on sensory attributes., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Olive oil application in the cosmetic industry may be extended by its ozonation, bringing about new oil properties and increased stability. Olive oil treated with 0.04 mole O 3 or 0.10 mole O 3 per 100 g oil was subjected to chemical parameters evaluation and composition scrutinizing by gas chromatography-mass spectrometry (GC-MS) and headspace solid-phase microextraction (HS-SPME) GC-MS analysis. The biological activity of refined and ozonated oil included their antimicrobial properties by the agar diffusion method and cytotoxicity by the MTT assay towards two normal (LLC-PK1, HaCaT) and two cancerous (Caco-2, HeLa) cell lines. The oils served as the basis in cosmetic emulsions. The chosen organoleptic features, preservative efficacy in a challenge test, and persistency during six months of these formulations were assessed. However, the ozonation of the olive oil resulted in a decrease in unsaturated acids; several additional compounds were detected in the ozonated oil, which positively affect the physicochemical, sensory, and functional properties of cosmetic emulsions. Emulsions based on the ozonated olive oil retain their properties longer compared to emulsions based on the refined olive oil. Ozonated oil treated with 0.10 mole O 3 /100 g oil allowed increasing the shelf life of the non-preserved formulation up to six months. A weak inhibitory effect against Candida albicans and Aspergillus brasiliensis was also demonstrated for this emulsion in the challenge test. Moreover, an interesting aroma, slightly enhanced antimicrobial activity against Escherichia coli , Staphylococcus aureus , C. albicans , A. brasiliensis, and a lack of cytotoxicity at concentrations 625 µg mL -1 make the ozonated olive oil a promising raw material for the cosmetics and pharmaceutical industries.

Biological control against microbial infections has a great potential as an alternative approach instead of fungicidal chemicals, which can cause environmental pollution. The pigment producer Metschnikowia andauensis belongs to the antagonistic yeasts, but details of the mechanism by which it inhibits growth of other microbes are less known. Our results confirmed its antagonistic capacity on other yeast species isolated from fruits or flowers and demonstrated that the antagonistic capacity was well correlated with the size of the red pigmented zone. We have isolated and characterized its red pigment, which proved to be the iron chelating pulcherrimin. Its production was possible even in the presence of 0.05 mg/ml copper sulphate, which is widely used in organic vineyards because of its antimicrobial properties. Production and localisation of the pulcherrimin strongly depended on composition of the media and other culture factors. Glucose, galactose, disaccharides and the presence of pectin or certain amino acids clearly promoted pigment production. Higher temperatures and iron concentration decreased the diameter of red pigmented zones. The effect of pH on pigment production varied depending of whether it was tested in liquid or solid media. In addition, our results suggest that other mechanisms besides the iron depletion of the culture media may contribute to the antagonistic capacity of M. andauensis.

In this study, it was aimed to develop a novel disinfectant from various essential oils containing active components with antimicrobial activity. The mixture of oregano, cinnamon and clove oils (1 : 1 : 1) with 10% oil concentration (SOM) was used as potential disinfectant on various areas and showed the highest antimicrobial activity among oil combinations tested. SOM reduced the numbers of total mesophilic aerobic bacteria (TMAB; 2·27 log CFU per 25 cm 2 ) and Escherichia coli (4·60 log CFU per 25 cm 2 ) under the detection limits. Application of SOM (1, 2, 3, 4 and 6%) into incubators reduced TMAB and mould-yeast counts of incubator air by 82·9 and 100% respectively. SOM application (3%) into ambient air also reduced its TMAB and mould-yeast counts by 92 and 84·6% respectively. While ethanol is commonly used for the disinfection of environments, equipment and surfaces, SOM is an important alternative that may also be used for the disinfection of various surfaces as well as air., (© 2020 The Society for Applied Microbiology.)

Androgens, both steroidal and nonsteroidal in nature, are among the most commonly misused substances in competitive sports. Their recognized anabolic and performance enhancing effects through short- and long-term physiological adaptations make them popular. Androgens exist as natural steroids, or are chemically synthesized as anabolic androgenic steroids (AAS) or selective androgen receptor modulators (SARMs). In order to effectively detect misuse of androgens, targeted strategies are used. These targeted strategies rely heavily on mass spectrometry, and detection requires prior knowledge of the targeted structure and its metabolites. Although exquisitely sensitive, such approaches may fail to detect novel structures that are developed and marketed. A nontargeted approach to androgen detection involves the use of cell-based in vitro bioassays. Both yeast and mammalian cell androgen bioassays demonstrate a clear ability to detect AAS and SARMS, and if paired with high resolution mass spectrometry can putatively identify novel structures. In vitro cell bioassays are successfully used to characterize designer molecules and to detect exogenous androgens in biological samples. It is important to continue to develop new and effective detection approaches to prevent misuse of designer androgens, and in vitro bioassays represent a potential solution to nontargeted detection strategies., (© 2021 John Wiley & Sons, Ltd.)

In our screening program for new biologically active compounds, a new polyene macrolide, lavencidin (1), along with known compound RKGS-A2215A (2), was isolated from the fermentation broth of Streptomyces lavendulae FRI-5 by changing the composition of liquid medium normally used for the strain. Their structures were elucidated by spectral methods (high-resolution fast-atom bombardment mass spectrometry (HRFABMS) and nuclear magnetic resonance (NMR)). Compound 1 includes a conjugated pentaene moiety together with six hydroxy groups and a carboxylic acid as a side chain. Lavencidin (1) showed moderate growth-inhibitory activity against yeast and was cytotoxic against human cancer cell lines with low-micromolar IC 50 values.

Local administration of antiseptics is required to prevent and fight against biofilm-based infections of chronic wounds. One of the methods used for delivering antiseptics to infected wounds is the application of dressings chemisorbed with antimicrobials. Dressings made of bacterial cellulose (BC) display several features, making them suitable for such a purpose. This work aimed to compare the activity of commonly used antiseptic molecules: octenidine, polyhexanide, povidone-iodine, chlorhexidine, ethacridine lactate, and hypochlorous solutions and to evaluate their usefulness as active substances of BC dressings against 48 bacterial strains (8 species) and 6 yeast strains (1 species). A silver dressing was applied as a control material of proven antimicrobial activity. The methodology applied included the assessment of minimal inhibitory concentrations (MIC) and minimal biofilm eradication concentration (MBEC), the modified disc-diffusion method, and the modified antibiofilm dressing activity measurement (A.D.A.M.) method. While in 96-well plate-based methods (MIC and MBEC assessment), the highest antimicrobial activity was recorded for chlorhexidine, in the modified disc-diffusion method and in the modified A.D.A.M test, povidone-iodine performed the best. In an in vitro setting simulating chronic wound conditions, BC dressings chemisorbed with polyhexanide, octenidine, or povidone-iodine displayed a similar or even higher antibiofilm activity than the control dressing containing silver molecules. If translated into clinical conditions, the obtained results suggest high applicability of BC dressings chemisorbed with antiseptics to eradicate biofilm from chronic wounds.

Yeasts are the leading cause of spoilage in yogurt. Considering the high demand from consumers to use natural products as an alternative to additives, essential oils (EOs) could be a promising solution to guarantee high microbiological standards. The present study highlighted the in vitro antifungal potential of cinnamon, ginger, lemongrass, mandarin, orange, lemon and lime EOs against spoilage yeasts isolated from yogurts prepared with pasteurized buffalo milk. A total of 74 isolates represented by 14 different species of Candida, Rhodotorula, Debaryomyces, Kluyveromyces and Yarrowia genera were subjected to a disc diffusion assay, showing lemongrass EO to have the highest antifungal activity (40.97 ± 9.86 mm), followed by cinnamon (38.46 ± 6.59 mm) and orange (12.00 ± 4.52 mm) EOs. Yarrowia lipolytica was less susceptible to lemongrass EO than Candida sake and Yarrowia deformans isolates. Ginger EO exhibited the lowest efficacy. A minimum inhibitory concentration (MIC) assay showed the ability of lemongrass and cinnamon EOs to inhibit the growth of all selected isolates at concentrations between ≤0.31 and 1.25 μL/mL. Therefore, for the first time, the two best-performing EOs (lemongrass and cinnamon) based on in vitro assays were assessed for their potential roles as preservatives in an in vivo yogurt model prepared at the laboratory scale. Since some limitations, such as the inhibition of lactic acid bacteria by cinnamon EO, consequently leading to fermentation failure as well as species-specific antifungal activity of lemongrass EO, were observed, further studies are needed to explore the possibility of using a slightly higher concentration of lemongrass EO and/or combinations of different EOs and/or their components. Finally, since yogurt spoilage could also be prevented by correct sanitation procedures of the production environment, the sanitizers commonly used in the food industry were tested against all isolates, showing the high efficiency of alcohol-based sanitizers and the ineffectiveness of chlorine-based sanitizers., (Copyright © 2021 Elsevier B.V. All rights reserved.)

In a previous article, we reported on the higher toxicity of silver(I) complexes of miconazole [Ag(MCZ) 2 NO 3 ( 1 )] and [Ag(MCZ) 2 ClO 4 ( 2 )] in HepG2 tumor cells compared to the corresponding salts of silver, miconazole and cisplatin. Here, we present the synthesis of two silver(I) complexes of miconazole containing two new counter ions in the form of Ag(MCZ) 2 X (MCZ = 1-[2-(2,4-dichlorobenzyloxy)-2-(2,4-dichlorophenyl)ethyl]-1H-imidazole]; X = BF 4 - ( 3 ), SbF 6 - ( 4 )). The novel silver(I) complexes were characterized by elemental analysis, 1 H NMR, 13 C NMR and infrared (IR) spectroscopy, electrospray ionization (ESI)-MS spectrometry and X-ray-crystallography. In the present study, the antimicrobial activity of all obtained silver(I) complexes of miconazole against six strains of Gram-positive bacteria, five strains of Gram-negative bacteria and yeasts was evaluated. The results were compared with those of a silver sulfadiazine drug, the corresponding silver salts and the free ligand. Silver(I) complexes exhibited significant activity against Gram-positive bacteria, which was much better than that of silver sulfadiazine and silver salts. The highest antimicrobial activity was observed for the complex containing the nitrate counter ion. All Ag(I) complexes of miconazole resulted in much better inhibition of yeast growth than silver sulfadiazine, silver salts and miconazole. Moreover, the synthesized silver(I) complexes showed good or moderate activity against Gram-negative bacteria compared to the free ligand.

Microbial resistance to available drugs is a growing health threat imposing the need for the development of new drugs. The scaffold of plant defensins, including their γ-cores, are particularly good candidates for drug design. This work aimed to improve the antifungal activity of a previous design peptide, named A 36 , 42 , 44 γ 32-46 VuDef (for short DD) against yeasts by altering its biochemical parameters. We explore the correlation of the biological activity and structure of plant defensins and compared their primary structures by superimposition with VuDef 1 and DD which indicated us the favorable position and the amino acid to be changed. Three new peptides with modifications in charge, hydrophobicity (RR and WR) and chirality (D-RR) were designed and tested against pathogenic yeasts. Inhibition was determined by absorbance. Viability of mammalian cells was determined by MTT. The three designed peptides had better inhibitory activity against the yeasts with better potency and spectrum of yeast species inhibition, with low toxicity to mammalian cells. WR, the most hydrophobic and cationic, exhibited better antifungal activity and lower toxicity. Our study provides experimental evidence that targeted changes in the primary structure of peptides based on plant defensins γ-core primary structures prove to be a good tool for the synthesis of new compounds that may be useful as alternative antifungal drugs. The method described did not have the drawback of synthesis of several peptides, because alterations are guided. When compared to other methods, the design process described is efficient and viable to those with scarce resources.

Although the mitochondrial permeability transition pore (PTP) is presumably formed by either ATP synthase or the ATP/ADP carrier (AAC), little is known about their differential roles in PTP activation. We explored the role of AAC and ATP synthase in PTP formation in Saccharomyces cerevisiae using bisindolylpyrrole (BP), an activator of the mammalian PTP. The yeast mitochondrial membrane potential, as indicated by tetramethylrhodamine methyl ester signals, dissipated over 2-4 h after treatment of cells with 5 μM BP, which was sensitive to cyclosporin A (CsA) and Cpr3 deficiency and blocked by porin1/2 deficiency. The BP-induced depolarization was inhibited by a specific AAC inhibitor, bongkrekate, and consistently blocked in a yeast strain lacking all three AACs, while it was not affected in the strain with defective ATP synthase dimerization, suggesting the involvement of an AAC-associated pore. Upon BP treatment, isolated yeast mitochondria underwent CsA- and bongkrekate-sensitive depolarization without affecting the mitochondrial calcein signals, indicating the induction of a low conductance channel. These data suggest that, upon BP treatment, yeast can form a porin1/2- and Cpr3-regulated PTP, which is mediated by AACs but not by ATP synthase dimers. This implies that yeast may be an excellent tool for the screening of PTP modulators.

Proguanil in combination with its synergistic partner atovaquone has been used for malaria treatment and prophylaxis for decades. However its mode of action is not fully understood. Here we used yeast to investigate its activity. Proguanil inhibits yeast growth, causes cell death and acts in synergy with atovaquone. It was previously proposed that the drug would target the system that maintains the mitochondrial membrane potential when the respiratory chain is inhibited. However our data did not seem to validate that hypothesis. We proposed that proguanil would not have a specific target but accumulate in the mitochondrial to concentrations that impair multiple mitochondrial functions leading to cell death. Selection and study of proguanil resistant mutants pointed towards an unexpected resistance mechanism: the decrease of CoQ level, which possibly alters the mitochondrial membrane properties and lowers proguanil intramitochondrial level., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Chlorpyrifos (CP) is one of the organophosphate insecticides most used worldwide today. Although the main target organ for CP is the nervous system triggering predominantly neurotoxic effects, it has suggested other mechanisms of action as cytotoxicity and endocrine disruption. The risk posed by the pesticide metabolites on non-target organisms is increasingly recognized by regulatory agencies and natural resource managers. In the present study, cytotoxicity and estrogenic activity of CP, and its principal metabolite 3,5,6-trichloro-2-pyridinol (TCP) have been evaluated by in vitro assays, using two mammalian cell lines (HEK293 and N2a), and a recombinant yeast. Results indicate that TCP is more toxic than CP for the two cell lines assayed, being N2a cells more sensitive to both compounds. Both compounds show a similar estrogenic activity being between 2500 and 3000 times less estrogenic than 17β-estradiol. In order to find new toxicity measurement models, yeasts isolated from marine sediments containing CP residues have been tested against CP and TCP by cell viability assay. Of the 12 yeast strains tested, 6 of them showed certain sensitivity, and a concentration-dependent response to the tested compounds, so they could be considered as future models for toxicity tests, although further investigations and proves are necessary.

In the food industry, food spoilage is a real issue that can lead to a significant amount of waste. Although current preservation techniques are being applied to reduce the occurrence of spoilage microorganisms, the problem persists. Food spoilage yeast are part of this dilemma, with common spoilers such as Zygosaccharomyces , Kluyveromyces , Debaryomyces and Saccharomyces frequently encountered. Antimicrobial peptides derived from plants have risen in popularity due to their ability to reduce spoilage. This study examines the potential application of a synthetic defensin peptide derived from barley endosperm. Its inhibitory effect against common spoilage yeasts, its mechanisms of action (membrane permeabilisation and overproduction of reactive oxygen species), and its stability in different conditions were characterised. The safety of the peptide was evaluated through a haemolysis and cytotoxicity assay, and no adverse effects were found. Both assays were performed to understand the effect of the peptide if it were to be consumed. Its ability to be degraded by a digestive enzyme was also examined for its safety. Finally, the peptide was successfully applied to different beverages and maintained the same inhibitory effects in apple juice as was observed in the antiyeast assays, providing further support for its application in food preservation.

We have isolated a filamentous fungus that actively secretes a pigmented exudate when growing on agar plates. The fungus was identified as being a strain of Epicoccum nigrum . The fungal exudate presented strong antifungal activity against both yeasts and filamentous fungi, and inhibited the germination of fungal spores. The chemical characterization of the exudate showed that the pigmented molecule presenting antifungal activity is the disalt of epipyrone A-a water-soluble polyene metabolite with a molecular mass of 612.29 and maximal UV-Vis absorbance at 428 nm. This antifungal compound showed excellent stability to different temperatures and neutral to alkaline pH.

Kpkt is a yeast killer toxin, naturally produced by Tetrapisispora phaffii, with possible applications in winemaking due to its antimicrobial activity on wine-related yeasts including Kloeckera/Hanseniaspora, Saccharomycodes and Zygosaccharomyces. Here, Kpkt coding gene was expressed in Komagataella phaffii (formerly Pichia pastoris) and the bioreactor production of the recombinant toxin (rKpkt) was obtained. Moreover, to produce a ready-to-use preparation of rKpkt, the cell-free supernatant of the K. phaffii recombinant killer clone was 80-fold concentrated and lyophilized. The resulting preparation could be easily solubilized in sterile distilled water and maintained its killer activity for up to six months at 4 °C. When applied to grape must, it exerted an extensive killer activity on wild wine-related yeasts while proving compatible with the fermentative activity of actively growing Saccharomyces cerevisiae starter strains. Moreover, it displayed a strong microbicidal effect on a variety of bacterial species including lactic acid bacteria and food-borne pathogens. On the contrary it showed no lethal effect on filamentous fungi and on Ceratitis capitata and Musca domestica, two insect species that may serve as non-mammalian model for biomedical research. Based on these results, bioreactor production and lyophilization represent an interesting option for the exploitation of this killer toxin that, due to its spectrum of action, may find application in the control of microbial contaminations in the wine and food industries., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aim of this research is to verify the yeast species isolated from oral mucosa in street mixed-breed dogs and to determine the antifungal profiles. After capturing and sedating the animals, oral mucosa samples were collected from fifty dogs and the materials were inoculated on Sabouraud dextrose agar with chloramphenicol. Forty-three yeast strains were isolated and identified trough the API-20C AUX method. Thirty-seven (86.1%) of the yeasts belonged to genus Candida, five (11.6%) to genus Trichosporon and only one strain (2.3%) to genus Malassezia. The sensitivity profiles to anifungals (amphotericin B, itraconanole, ketoconazole, fluconazole and variconazole) were determined through Etest® method. This study found resistance of some yeasts to amphotericin B and a good susceptibility to voriconazole and ketoconazole. Some of these antifungals are used in veterinary medical practice. This research is the first investigation on street mixed-breed dogs regarding yeast identifications and antifungals profiles., (Copyright © 2020. Published by Elsevier Masson SAS.)

Isavuconazole is the last antifungal agent approved by the FDA and available for treatment of fungal infections. In the present study, the in vitro activity of isavuconazole against several yeasts was investigated. Two hundred forty-six isolates were included: 64 Candida albicans, 53 Candida parapsilosis sensu stricto, 48 Cryptococcus neoformans species complex, 27 C. glabrata sensu stricto, 17 C. lusitaniae, 17 C. tropicalis, 5 C. orthopsilosis, 4 C. krusei, 3 C. guilliermondii sensu stricto, 3 C. pelliculosa, 2 C. dubliniensis, 1 C. auris, 1 C. fermentati and 1 Trichosporon asahii. All isolates were recovered from clinical isolates from Chile, being 221 from hemoculture, 22 from cerebrospinal fluid, 1 pleural fluid, and 1 from tissue culture. The minimal inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs) of isavuconazole were determined. Isavuconazole demonstrated good in vitro activity against all species tested. MIC90 values and MFC ranges of isavuconazole for Candida albicans were 0.03 mg/L and 0.03- > 16 mg/L respectively. Non-Candida albicans species isolates were inhibited by ≤ 1 mg/L, with MFC ranges from < 0.03- > 16 mg/L. Also, isavuconazole was active against the non-Candida yeasts, being inhibited with MIC values ≤ 0.06 mg/L. Isavuconazole has exhibited potent in vitro activity against clinical isolates of Candida spp., Cryptococcus neoformans species complex, and other clinical yeast in Chile. Despite the results obtained in the present work, additional clinical studies are necessary to verify the level of efficacy of this azole.

Microbiological contamination by spoilage yeasts species are frequent during winemaking, and biological control using antagonistic yeasts is considered a more beneficial alternative to conventional synthetic antimicrobials. Saccharomyces eubayanus killer toxin (SeKT) was produced and purified in a synthetic optimized medium. Purification procedure allowed the identification of SeKT as protein with an apparent molecular mass of 70 kDa and activity at physicochemical conditions suitable for winemaking process. Purified SeKT reduced the levels of volatile phenols produced by the spoilage yeasts Brettanomyces bruxellensis, Pichia membranifaciens, Meyerozyma guilliermondii and Pichia manshurica in wine-like medium. The putative mode of action of SeKT on sensitive yeast strains comprises cell wall disruption through β-glucanase and chitinase activities as well as necrotic and apoptotic death in a toxin dose dependent manner. Thus, SeKT appears to be a promising biocontrol agent against spoilage yeasts during wine aging and storing., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

To create a broad-spectrum peptide biocide, we synthesized 45 analogs of antimicrobial peptide indolicidin (H-Ile-Leu-Pro-Trp-Lys-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-NH 2 ). Among them the peptides H-Ile-Leu-Pro-(2-Me)Phe-Lys-(2-Me)Phe-Pro-(2-Me)Phe-(2-Me)Phe-Pro-(2-Me)Phe-Arg-Arg-NH 2 and HN 2 -(CH 2 ) 10 -Ile-Leu-Pro-D-Phe-Lys-D-Phe-Pro-D-Phe-D-Phe-Pro-D-Phe-Arg-Arg-NH 2 have the broadest spectrum of antimicrobial activity and the lowest hemolytic activity. They are active against all 11 tested strains of Gram-positive bacteria, Gram-negative bacteria and fungi with MIC 50 from 0.9 to 6.1 μg/ml (0.5 to 3.2 μM), being up to 3 times more active than indolicidin, and are at least 1.8 times less hemolytically active than indolicidin (reached the detection limit). These peptides are patented and could be used for further drug development as antimicrobials., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Aims: The objective of this study was to evaluate the potential of hexanoic acid (Hex) as a silage additive., Methods and Results: The effect of Hex, Lactobacillus plantarum (Lp) and their combination (Hex + Lp) on the aerobic stability of napier grass silage was investigated. Napier grass was ensiled without additives (C) or with Lp, Hex or Hex + Lp for 60 days followed by 7 days of aerobic exposure. After 60 days of ensiling, the Lp silage had the lowest pH and the highest lactic acid (LA) concentration among all silage, whereas the highest water-soluble carbohydrates (WSC) content was observed in Hex + Lp silage, followed by Hex silage. After 60 days of ensiling, the population of yeasts in C and Lp silages was > 3·0 log 10 CFU per g FW, while that of Hex and Hex + Lp was <2·0 log 10 CFU per g FW. During aerobic exposure, the pH in the Lp and C silages increased (P < 0·05) above 7·0 on day 5. The pH of Hex silage was the lowest among all silages on day 3 and 5, followed by a significant (P < 0·05) increase until 7 days of aerobic exposure. There were no significant changes in pH and AA of Hex + Lp silage over the duration of aerobic exposure. The concentrations of LA in C, Lp and Hex silages decreased while that of Hex + Lp silage remained stable after 3 days of aerobic exposure. The Hex delayed the decline of WSC contents and the increase in yeasts over the aerobic exposure period., Conclusion: Addition of Hex (97 h) and Hex + Lp (>168 h) improved aerobic stability (P < 0·05) as compared to the control (83 h)., Significance and Impact of the Study: Hex or in combination with L. plantarum inhibited the proliferation of yeasts during aerobic exposure of napier grass. Thus, Hex is an alternative antifungal additive to improve aerobic stability., (© 2020 The Society for Applied Microbiology.)

In natural environments, microorganisms form microbial aggregates called biofilms able to adhere to a multitude of different surfaces. Yeasts make no exception to this rule, being able to form biofilms in a plethora of environmental niches. In food realms, yeast biofilms may cause major problems due to their alterative activities. In addition, yeast biofilms are tenacious structures difficult to eradicate or treat with the current arsenal of antifungal agents. Thus, much effort is being made to develop novel approaches to prevent and disrupt yeast biofilms, for example through the use of natural antimicrobials or small molecules with both inhibiting and dispersing properties. The aim of this review is to provide a synopsis of the most recent literature on yeast biofilms regarding: (i) biofilm formation mechanisms; (ii) occurrence in food and in food-related environments; and (iii) inhibition and dispersal using natural compounds, in particular.

Background: Yeasts, which are ubiquitous in agroecosystems, are known to degrade various xenobiotics. The aim of this study was to analyze the effect of fungicides on the abundance of natural yeast communities colonizing winter wheat leaves, to evaluate the sensitivity of yeast isolates to fungicides in vivo, and to select yeasts that degrade propiconazole., Results: Fungicides applied during the growing season generally did not affect the counts of endophytic yeasts colonizing wheat leaves. Propiconazole and a commercial mixture of flusilazole and carbendazim decreased the counts of epiphytic yeasts, but the size of the yeast community was restored after 10 days. Epoxiconazole and a commercial mixture of fluoxastrobin and prothioconazole clearly stimulated epiphyte growth. The predominant species isolated from leaves were Aureobasidium pullulans and Rhodotorula glutinis. In the disk diffusion test, 14 out of 75 yeast isolates were not sensitive to any of the tested fungicides. After 48 h of incubation in an aqueous solution of propiconazole, the Rhodotorula glutinis Rg 55 isolate degraded the fungicide in 75%. Isolates Rh. glutinis Rg 92 and Rg 55 minimized the phytotoxic effects of propiconazole under greenhouse conditions. The first isolate contributed to an increase in the dry matter content of wheat seedlings, whereas the other reduced the severity of chlorosis., Conclusion: Not sensitivity of many yeast colonizing wheat leaves on the fungicides and the potential of isolate Rhodotorula glutinis Rg 55 to degrade of propiconazole was established. Yeast may partially eliminate the ecologically negative effect of fungicides.

Introduction: Antimicrobial peptides are proving to be promising lead compounds for therapeutics. The major disadvantage of antimicrobial peptides is their proteolytic instability in the body, with half-lives averaging less than an hour. Peptoids, or N-substituted glycines, have emerged as a promising field of peptidomimetics by retaining the beneficial properties of antimicrobial peptides while improving their stability., Methods: This study evaluated peptoid derivatives of ultra-short lipophilic antimicrobial peptides, comparing their potency side-by-side with the most prevalent multidrug-resistant bacteria (ESKAPE) and yeast pathogens (Candida albicans and Cryptococcus neoformans)., Results: Both peptide and peptoid counterparts were most effective against Gram-positive bacteria with minimum inhibitory concentration (MIC) values as low as 1.6 and 6.3 µg/mL, respectively. In general, peptides retained better antimicrobial activity than their peptoid counterparts; however, certain peptoids proved to be more effective than peptides against Gram-negative bacteria. For example, peptoid MG10 displayed an MIC of 6.3 µg/mL against Pseudomonas aeruginosa compared with the peptide counterpart with an MIC of 100 µg/mL. All tested compounds were more potent against Cryptococcus neoformans compared with Candida albicans. Cytotoxicity analysis indicated that peptoids were generally slightly less toxic than their peptide counterparts. Additionally, trypsin rapidly degraded one of the evaluated peptides, while having no effect on comparable peptoids, demonstrating the proteolytic stability of peptoids., Conclusion: These results indicate that direct conversion of lipopeptides to lipopeptoids can result in compounds with comparable antimicrobial activity, favorable mammalian cell toxicity, and excellent proteolytic stability., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Along with swift economic evolution and continuous amelioration of lifestyle, people at present are paying more attention to health issues. Synthetic drugs will be compensated with other natural ones that belong to natural origin. Plants have always been considered as sources of several compounds that are used in many fields, especially human and animal health, starting from boosting immunity to the treatment of infectious diseases caused by some pathogenic microbes such as bacteria, fungi as well as viruses. This study aimed to incorporate some types of plants within the antimicrobial portfolio through the examination of different six plants which were Cichorium intybus, Cinnamomum camphora, Commiphora myrrha, Foeniculum vulgare, Nerium oleander, and Spartium junceum. As well, attempting to identify the active constituents of their extracts using GC-MS., Materials and Methods: All selected plants were analyzed to determine their phytochemical composition such as phenolics, alkaloids, flavonoids, terpenoids, and so on. The extraction step was done by sophisticated equipment called supercritical fluid extractor SFE through adjustment of specific conditions include temperature, time, flow rate and pressure to change the behavior of CO 2 . Testing the antimicrobial activity of each plant extract via agar well diffusion method through the formation of clear zones against a wide range of test microorganisms including both Gram-positive and Gram-negative bacteria as well as yeasts. Finally, attempting to primarily identify the constituents of each plant extract using GC-MS., Results and Discussion: The crude extract of F. vulgare showed the highest potency against C. albicans, E. faecalis and S. typhimurium, it contains some unique compounds such as squalene, eugenol and isoeugenol while, Extract of C. intybus showed a moderate activity especially against C. lipolytica and MRSA and it includes Vitamin A like compound which indicates antioxidant property., Conclusion: Conclusively, fennel gave a promising result as a good wide spectrum antimicrobial agent because it contains some compounds act as antimicrobial agents such as eugenol which was used as food preservatives in addition to squalene which acts as an antioxidant and antimycotic agent so, it will be useful especially while it was used in highly purified form excluding all undesirable subcomponents.