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1.
Probiotics Antimicrob Proteins ; 14(3): 595-601, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35451745

RESUMO

Mycocins are substances that have the potential to affect other sensitive yeasts or microorganisms. Wickerhamomyces anomalus is a yeast that produces mycocins that have great biotechnological potential, being highly competitive in many habitats, as it is adaptable to a wide range of environmental conditions. Thus, they are targets for studies in different areas, including the environment, industry, and medical sciences. Yeasts of the genus Candida are of great importance due to the high frequency with which they colonize and infect the human host. Yeast infections are often difficult to treat due to the acquisition of resistance against antifungals, leading to studies focusing in new treatment alternatives. This work aims to verify the inhibition of Candida albicans isolated from vaginal secretion by mycocins produced by Wickerhamomyces anomalus. Tests were carried out in solid medium and microdilution tests, where mycocins proved to be efficient in inhibiting the growth of C. albicans, hemolysis, and irritation in an organotypic model, which showed that the mycocins produced by W. anomalus are safe and non-irritating. Thus, the results of this work can provide scientific evidence for the application of mycocins in the production of new antifungal alternatives.


Assuntos
Candida albicans , Saccharomycetales , Antifúngicos/farmacologia , Candida , Feminino , Humanos , Leveduras
2.
Appl Environ Microbiol ; 88(8): e0203021, 2022 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-35389250

RESUMO

Yeast killer toxins are widely distributed in nature, conferring a competitive advantage to the producer yeasts over nonkiller ones when nutrients are scarce. Most of these toxins are encoded on double-stranded RNAs (dsRNAs) generically called M. L-A members of the viral family Totiviridae act as helper viruses to maintain M, providing the virion proteins that separately encapsidate and replicate L-A and M genomes. M genomes are organized in three regions, a 5' region coding the preprotoxin, followed by an internal poly(A) stretch and a 3' noncoding region. In this work, we report the characterization of K74 toxin encoded on M74 dsRNA from Saccharomyces paradoxus Q74.4. In M74, there is a 5' upstream sequence of 141 nucleotides (nt), which contains regulatory signals for internal translation of the preprotoxin open reading frame (ORF) at the second AUG codon. The first AUG close to the 5' end is not functional. For K74 analysis, M74 viruses were first introduced into laboratory strains of Saccharomyces cerevisiae. We show here that the mature toxin is an α/ß heterodimer linked by disulfide bonds. Though the toxin (or preprotoxin) confers immunity to the carrier, cells with increased K74 loads have a sick phenotype that may lead to cell death. Thus, a fine-tuning of K74 production by the upstream regulatory sequence is essential for the host cell to benefit from the toxin it produces and, at the same time, to safely avoid damage by an excess of toxin. IMPORTANCE Killer yeasts produce toxins to which they are immune by mechanisms not well understood. This self-immunity, however, is compromised in certain strains, which secrete an excess of toxin, leading to sick cells or suicidal phenotypes. Thus, a fine-tuning of toxin production has to be achieved to reach a balance between the beneficial effect of toxin production and the stress imposed on the host metabolism. K74 toxin from S. paradoxus is very active against Saccharomyces uvarum, among other yeasts, but an excess of toxin production is deleterious for the host. Here, we report that the presence of a 5' 141-nt upstream sequence downregulates K74 toxin precursor translation, decreasing toxin levels 3- to 5-fold. Thus, this is a special case of translation regulation performed by sequences on the M74 genome itself, which have been presumably incorporated into the viral RNA during evolution for that purpose.


Assuntos
RNA de Cadeia Dupla , Saccharomyces cerevisiae , Humanos , Fatores Matadores de Levedura/genética , RNA de Cadeia Dupla/genética , RNA de Cadeia Dupla/metabolismo , Saccharomyces , Saccharomyces cerevisiae/genética , Regiões não Traduzidas
3.
Microorganisms ; 11(1)2022 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-36677329

RESUMO

(1) Background: Wine yeast research offers the possibility of isolating new strains with distinct metabolic properties due to the geographical location of the vineyard and the processes used in winemaking. Our study deals with the isolation and identification of six yeasts from spontaneously fermented wine wort from Romania and their characterization as new potential starter culture for traditional beverages, for food industry or biomedicine. (2) Materials and methods: The isolates were identified using conventional taxonomy tests, phenotypic phylogeny analysis (Biolog YT), MALDI-TOF mass spectrometry, PCR-RFLP, and sequencing of the ITS1-5,8S-ITS2 rDNA region. The capacity of the yeasts to grow under thermal, ionic, and osmotic stress was determined. The safe status was confirmed by testing virulence and pathogenicity factors. Assays were performed in order to evaluate the growth inhibition of Candida strains and determine the antimicrobial mechanism of action. (3) Results and discussions: The yeast isolates were identified as belonging to the Metschinikowia, Hanseniaspora, Torulaspora, Pichia, and Saccharomyces genera. All the isolates were able to develop under the tested stress conditions and were confirmed as safe. With the exception of S. cerevisiae CMGB-MS1-1, all the isolates showed good antimicrobial activity based on competition for iron ions or production of killer toxins. (4) Conclusions: The results revealed the resistance of our yeasts to environmental conditions related to industrial and biomedical applications and their high potential as starter cultures and biocontrol agents, respectively.

4.
Toxins (Basel) ; 13(9)2021 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-34564619

RESUMO

Yeasts can have additional genetic information in the form of cytoplasmic linear dsDNA molecules called virus-like elements (VLEs). Some of them encode killer toxins. The aim of this work was to investigate the prevalence of such elements in D. hansenii killer yeast deposited in culture collections as well as in strains freshly isolated from blue cheeses. Possible benefits to the host from harboring such VLEs were analyzed. VLEs occurred frequently among fresh D. hansenii isolates (15/60 strains), as opposed to strains obtained from culture collections (0/75 strains). Eight new different systems were identified: four composed of two elements and four of three elements. Full sequences of three new VLE systems obtained by NGS revealed extremely high conservation among the largest molecules in these systems except for one ORF, probably encoding a protein resembling immunity determinant to killer toxins of VLE origin in other yeast species. ORFs that could be potentially involved in killer activity due to similarity to genes encoding proteins with domains of chitin-binding/digesting and deoxyribonuclease NucA/NucB activity, could be distinguished in smaller molecules. However, the discovered VLEs were not involved in the biocontrol of Yarrowia lipolytica and Penicillium roqueforti present in blue cheeses.


Assuntos
Queijo/virologia , Citoplasma/virologia , Debaryomyces/virologia , Micotoxinas/análise , Retroelementos
5.
Toxins (Basel) ; 13(9)2021 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-34564659

RESUMO

Possible implications and applications of the yeast killer phenomenon in the fight against infectious diseases are reviewed, with particular reference to some wide-spectrum killer toxins (KTs) produced by Wickerhamomyces anomalus and other related species. A perspective on the applications of these KTs in the medical field is provided considering (1) a direct use of killer strains, in particular in the symbiotic control of arthropod-borne diseases; (2) a direct use of KTs as experimental therapeutic agents; (3) the production, through the idiotypic network, of immunological derivatives of KTs and their use as potential anti-infective therapeutics. Studies on immunological derivatives of KTs in the context of vaccine development are also described.


Assuntos
Anti-Infecciosos/toxicidade , Anti-Infecciosos/uso terapêutico , Doenças Transmissíveis/tratamento farmacológico , Citotoxinas/uso terapêutico , Fatores Matadores de Levedura/toxicidade , Fatores Matadores de Levedura/uso terapêutico , Saccharomycetales/química , Humanos , Desenvolvimento de Vacinas
6.
Antimicrob Agents Chemother ; 65(7): e0245020, 2021 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-33972245

RESUMO

Compared to other species of Candida yeasts, the growth of Candida glabrata is inhibited by many different strains of Saccharomyces killer yeasts. The ionophoric K1 and K2 killer toxins are broadly inhibitory to all clinical isolates of C. glabrata from patients with recurrent vulvovaginal candidiasis, despite high levels of resistance to clinically relevant antifungal therapeutics.


Assuntos
Candida glabrata , Candidíase Vulvovaginal , Antifúngicos/farmacologia , Candida glabrata/genética , Candidíase Vulvovaginal/tratamento farmacológico , Farmacorresistência Fúngica/genética , Feminino , Humanos , Ionóforos , Testes de Sensibilidade Microbiana , Saccharomyces cerevisiae/genética
7.
Fungal Genet Biol ; 148: 103518, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33497840

RESUMO

Despite the interest on fungi as eukaryotic model systems, the molecular mechanisms regulating the fungal non-self-recognition at a distance have not been studied so far. This paper investigates the molecular mechanisms regulating the cross-talk at a distance between two filamentous fungi, Trichoderma gamsii and Fusarium graminearum which establish a mycoparasitic interaction where T. gamsii and F. graminearum play the roles of mycoparasite and prey, respectively. In the present work, we use an integrated approach involving dual culture tests, comparative genomics and transcriptomics to investigate the fungal interaction before contact ('sensing phase'). Dual culture tests demonstrate that growth rate of F. graminearum accelerates in presence of T. gamsii at the sensing phase. T. gamsii up-regulates the expression of a ferric reductase involved in iron acquisition, while F. graminearum up-regulates the expression of genes coding for transmembrane transporters and killer toxins. At the same time, T. gamsii decreases the level of extracellular interaction by down-regulating genes coding for hydrolytic enzymes acting on fungal cell wall (chitinases). Given the importance of fungi as eukaryotic model systems and the ever-increasing genomic resources available, the integrated approach hereby presented can be applied to other interactions to deepen the knowledge on fungal communication at a distance.


Assuntos
Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fungos/genética , Fungos/metabolismo , Transdução de Sinais , Parede Celular/metabolismo , Quitinases/genética , Fungos/citologia , Fusarium/genética , Fusarium/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação Fúngica da Expressão Gênica , Genômica/métodos , Hypocreales/genética , Hypocreales/metabolismo , Doenças das Plantas/microbiologia , Receptor Cross-Talk
8.
Med Mycol ; 58(8): 1102-1113, 2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-32196549

RESUMO

Candidiasis is a group of opportunistic infections caused by yeast of the genus Candida. The appearance of drug resistance and the adverse effects of current antifungal therapies require the search for new, more efficient therapeutic alternatives. Killer yeasts have aroused as suitable candidates for mining new antifungal compounds. Killer strains secrete antimicrobial proteins named killer toxins, with promissory antifungal activity. Here we found that the killer yeast Wickerhamomyces anomalus Cf20 and its cell-free supernatant (CFS) inhibited six pathogenic strains and one collection strain of Candida spp. The inhibition is mainly mediated by secreted killer toxins and, to a lesser extent, by volatile compounds such as acetic acid and ethyl acetate. A new large killer toxin (>180 kDa) was purified, which exerted 70-74% of the total CFS anti-Candida activity, and the previously described glucanase KTCf20 was inhibitory in a lesser extent as well. In addition, we demonstrated that Cf20 possesses the genes encoding for the ß-1,3-glucanases WaExg1 and WaExg2, proteins with extensively studied antifungal activity, particularly WaExg2. Finally, the 10-fold concentrated CFS exerted a high candidacidal effect at 37°C, completely inhibiting the fungal growth, although the nonconcentrated CFS (RCF 1) had very limited fungistatic activity at this temperature. In conclusion, W. anomalus Cf20 produces different low and high molecular weight compounds with anti-Candida activity that could be used to design new therapies for candidiasis and as a source for novel antimicrobial compounds as well.


Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Fatores Matadores de Levedura/farmacologia , Saccharomycetales/metabolismo , Compostos Orgânicos Voláteis/farmacologia , Antifúngicos/análise , Candida/crescimento & desenvolvimento , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/farmacologia , Relação Dose-Resposta a Droga , Fatores Matadores de Levedura/análise , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Compostos Orgânicos Voláteis/análise , Leveduras/classificação , Leveduras/metabolismo
9.
Antonie Van Leeuwenhoek ; 112(7): 965-973, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30671692

RESUMO

The juicing industry generates large amounts of waste that mostly lack commercial value and, in the absence of waste treatment policies, produces environmental pollution. Also, microbiological spoilage is a major concern in the wine industry and control tools are limited. Taking these challenges into account, agro-industrial waste coming from ultrafiltrated apple and pear juice were used to grow Saccharomyces eubayanus and to produce its killer toxin (SeKT). A Plackett-Burman screening was performed in order to optimize SeKT production in ultrafiltrated apple and pear juice. The optimized medium was characterized: 75% v/v WUJ, 0.5% m/v KH2PO4, 0.5% m/v MgSO4, 0.5% m/v (NH4)SO4, 0.5% g/L urea, 10% v/v glycerol and 0.1% v/v Triton X-100. SeKT produced in WUJ optimised medium was used to perform killer assays against wine spoilage yeasts and showed antagonistic activity against Brettanomyces bruxellensis, Pichia guilliermondii, Pichia manshurica and Pichia membranifaciens. Different inhibition percentages against spoilage species in a wine environment (49-69%) were detected and preserved for at least 48 h. For the first time, this work reports the ability of S. eubayanus to produce a killer toxin with potential use as a biocontrol tool in winemaking. Producing SeKT using agro-industrial waste as an alternative medium to cultivate S. eubayanus would have industrial, economic and ecological benefits.


Assuntos
Microbiologia Industrial/métodos , Resíduos Industriais/análise , Fatores Matadores de Levedura/metabolismo , Saccharomyces/metabolismo , Vinho/microbiologia , Meios de Cultura/química , Meios de Cultura/metabolismo , Fermentação , Microbiologia Industrial/instrumentação , Fatores Matadores de Levedura/farmacologia , Pichia/efeitos dos fármacos , Pichia/crescimento & desenvolvimento , Saccharomyces/química , Saccharomyces/genética , Resíduos/análise
10.
Toxins (Basel) ; 9(10)2017 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-29019944

RESUMO

Yeasts within the Saccharomyces sensu stricto cluster can produce different killer toxins. Each toxin is encoded by a medium size (1.5-2.4 Kb) M dsRNA virus, maintained by a larger helper virus generally called L-A (4.6 Kb). Different types of L-A are found associated to specific Ms: L-A in K1 strains and L-A-2 in K2 strains. Here, we extend the analysis of L-A helper viruses to yeasts other than S. cerevisiae, namely S. paradoxus, S. uvarum and S. kudriavzevii. Our sequencing data from nine new L-A variants confirm the specific association of each toxin-producing M and its helper virus, suggesting co-evolution. Their nucleotide sequences vary from 10% to 30% and the variation seems to depend on the geographical location of the hosts, suggesting cross-species transmission between species in the same habitat. Finally, we transferred by genetic methods different killer viruses from S. paradoxus into S. cerevisiae or viruses from S. cerevisiae into S. uvarum or S. kudriavzevii. In the foster hosts, we observed no impairment for their stable transmission and maintenance, indicating that the requirements for virus amplification in these species are essentially the same. We also characterized new killer toxins from S. paradoxus and constructed "superkiller" strains expressing them.


Assuntos
Fatores Matadores de Levedura/biossíntese , Saccharomyces/metabolismo , Saccharomyces/virologia , Totivirus/fisiologia , Sequência de Bases , DNA Complementar/genética , DNA Viral/genética , Interações Hospedeiro-Patógeno , Totivirus/genética
11.
J Appl Microbiol ; 122(3): 747-758, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27992098

RESUMO

AIMS: The control of the wine spoilage yeast Brettanomyces bruxellensis using biological methods such as killer toxins (instead of the traditional chemical methods, e.g. SO2 ) has been the focus of several studies within the last decade. Our previous research demonstrated that the killer toxins CpKT1 and CpKT2 isolated from the wine yeast Candida pyralidae were active and stable under winemaking conditions. In this study, we report the possible mode of action of CpKT1 on B. bruxellensis cells in red grape juice. METHODS AND RESULTS: Brettanomyces bruxellensis cells were exposed to CpKT1 either directly or through co-inoculation with C. pyralidae. This exposure yielded a temporary or permanent decline of the spoilage yeast population depending on the initial cell concentration. Scanning electron microscopy revealed cell surface abrasion while propidium iodide viability staining showed that CpKT1 caused plasma membrane damage on B. bruxellensis cells. Our data show that the exposure to CpKT1 resulted in increased levels of ß-glucan, suggesting a compensatory response of the sensitive cells. CONCLUSIONS: The toxin CpKT1 causes cell membrane and cell wall damage in B. bruxellensis. SIGNIFICANCE AND IMPACT OF THE STUDY: Candida pyralidae shows potential to be used as a biocontrol agent against B. bruxellensis in grape juice/wine.


Assuntos
Brettanomyces/efeitos dos fármacos , Candida/metabolismo , Parede Celular/efeitos dos fármacos , Micotoxinas/farmacologia , Brettanomyces/ultraestrutura , Parede Celular/ultraestrutura , Microbiologia de Alimentos , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Micotoxinas/isolamento & purificação , Propídio , Vitis/microbiologia , Vinho/microbiologia , Fermento Seco , beta-Glucanas/metabolismo
12.
Appl Environ Microbiol ; 83(4)2017 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-27940540

RESUMO

Saccharomyces cerevisiae killer strains secrete a protein toxin active on nonkiller strains of the same (or other) yeast species. Different killer toxins, K1, K2, K28, and Klus, have been described. Each toxin is encoded by a medium-size (1.5- to 2.3-kb) M double-stranded RNA (dsRNA) located in the cytoplasm. M dsRNAs require L-A helper virus for maintenance. L-A belongs to the Totiviridae family, and its dsRNA genome of 4.6 kb codes for the major capsid protein Gag and a minor Gag-Pol protein, which form the virions that separately encapsidate L-A or the M satellites. Different L-A variants exist in nature; on average, 24% of their nucleotides are different. Previously, we reported that L-A-lus was specifically associated with Mlus, suggesting coevolution, and proposed a role of the toxin-encoding M dsRNAs in the appearance of new L-A variants. Here we confirm this by analyzing the helper virus in K2 killer wine strains, which we named L-A-2. L-A-2 is required for M2 maintenance, and neither L-A nor L-A-lus shows helper activity for M2 in the same genetic background. This requirement is overcome when coat proteins are provided in large amounts by a vector or in ski mutants. The genome of another totivirus, L-BC, frequently accompanying L-A in the same cells shows a lower degree of variation than does L-A (about 10% of nucleotides are different). Although L-BC has no helper activity for M dsRNAs, distinct L-BC variants are associated with a particular killer strain. The so-called L-BC-lus (in Klus strains) and L-BC-2 (in K2 strains) are analyzed. IMPORTANCE: Killer strains of S. cerevisiae secrete protein toxins that kill nonkiller yeasts. The "killer phenomenon" depends on two dsRNA viruses: L-A and M. M encodes the toxin, and L-A, the helper virus, provides the capsids for both viruses. Different killer toxins exist: K1, K2, K28, and Klus, encoded on different M viruses. Our data indicate that each M dsRNA depends on a specific helper virus; these helper viruses have nucleotide sequences that may be as much as 26% different, suggesting coevolution. In wine environments, K2 and Klus strains frequently coexist. We have previously characterized the association of Mlus and L-A-lus. Here we sequence and characterize L-A-2, the helper virus of M2, establishing the helper virus requirements of M2, which had not been completely elucidated. We also report the existence of two specific L-BC totiviruses in Klus and K2 strains with about 10% of their nucleotides different, suggesting different evolutionary histories from those of L-A viruses.


Assuntos
Antifúngicos/metabolismo , Vírus Auxiliares/genética , Saccharomyces cerevisiae/virologia , Totivirus/genética , Capsídeo/metabolismo , Proteínas de Fusão gag-pol/genética , Produtos do Gene gag/genética , Mitocôndrias/genética , Saccharomyces cerevisiae/metabolismo , Vírus Satélites/genética
13.
Int J Food Microbiol ; 217: 94-100, 2016 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-26513248

RESUMO

Microbiological spoilage is a major concern throughout the wine industry, and control tools are limited. This paper addresses the identification and partial characterization of a new killer toxin from Torulaspora delbrueckii with potential biocontrol activity of Brettanomyces bruxellensis, Pichia guilliermondii, Pichia manshurica and Pichia membranifaciens wine spoilage. A panel of 18 different wine strains of T. delbrueckii killer yeasts was analysed, and the strain T. delbrueckii NPCC 1033 (TdKT producer) showed a significant inhibitory effect on the growth of all different spoilage yeasts evaluated. The TdKT toxin was then subjected to a partial biochemical characterization. Its estimated molecular weight was N30 kDa and it showed glucanase and chitinase enzymatic activities. The killer activity was stable between pH 4.2 and 4.8 and inactivated at temperature above 40 °C. Pustulan and chitin ­ but not other cell wall polysaccharides ­ prevented sensitive yeast cells from being killed by TdKT, suggesting that those may be the first toxin targets in the cell wall. TdKT provoked an increase in necrosis cell death after 3 h treatment and apoptotic cell death after 24 h showing time dependence in its mechanisms of action. Killer toxin extracts were active at oenological conditions, confirming their potential use as a biocontrol tool in winemaking.


Assuntos
Fatores Matadores de Levedura/farmacologia , Pichia/efeitos dos fármacos , Torulaspora/metabolismo , Vinho/microbiologia , Quitinases/metabolismo , Dextranase/metabolismo , Polissacarídeos Fúngicos/metabolismo , Fatores Matadores de Levedura/química , Fatores Matadores de Levedura/isolamento & purificação , Testes de Sensibilidade Microbiana , Temperatura , Torulaspora/patogenicidade
14.
Crit Rev Biotechnol ; 35(2): 222-34, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24102112

RESUMO

Killer toxins secreted by some yeast strains are the proteins that kill sensitive cells of the same or related yeast genera. In recent years, many new yeast species have been found to be able to produce killer toxins against the pathogenic yeasts, especially Candida albicans. Some of the killer toxins have been purified and characterized, and the genes encoding the killer toxins have been cloned and characterized. Many new targets including different components of cell wall, plasma membrane, tRNA, DNA and others in the sensitive cells for the killer toxin action have been identified so that the new molecular mechanisms of action have been elucidated. However, it is still unknown how some of the newly discovered killer toxins kill the sensitive cells. Studies on the killer phenomenon in yeasts have provided valuable insights into a number of fundamental aspects of eukaryotic cell biology and interactions of different eukaryotic cells. Elucidation of the molecular mechanisms of their action will be helpful to develop the strategies to fight more and more harmful yeasts.


Assuntos
Fatores Matadores de Levedura , Biotecnologia , Candida
15.
Microorganisms ; 3(4): 588-611, 2015 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-27682107

RESUMO

The yeasts constitute a large group of microorganisms characterized by the ability to grow and survive in different and stressful conditions and then to colonize a wide range of environmental and human ecosystems. The competitive traits against other microorganisms have attracted increasing attention from scientists, who proposed their successful application as bioprotective agents in the agricultural, food and medical sectors. These antagonistic activities rely on the competition for nutrients, production and tolerance of high concentrations of ethanol, as well as the synthesis of a large class of antimicrobial compounds, known as killer toxins, which showed clearly a large spectrum of activity against food spoilage microorganisms, but also against plant, animal and human pathogens. This review describes the antimicrobial mechanisms involved in the antagonistic activity, their applications in the processed and unprocessed food sectors, as well as the future perspectives in the development of new bio-drugs, which may overcome the limitations connected to conventional antimicrobial and drug resistance.

16.
Int J Food Microbiol ; 188: 83-91, 2014 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-25087208

RESUMO

Wine spoilage associated with Brettanomyces bruxellensis is a major concern for winemakers. An effective and reliable method to control the proliferation of this yeast is therefore of utmost importance. To achieve this purpose, sulphur dioxide (SO2) is commonly employed but the efficiency of this chemical compound is subject to wine composition and it can elicit allergic reactions in some consumers. Biological alternatives are therefore actively sought. The current study focused on identifying and characterizing killer toxins which are antimicrobial compounds that show potential in inhibiting B. bruxellensis in wine. Two killer toxins, CpKT1 and CpKT2, from the wine isolated yeast Candida pyralidae were identified and partially characterized. The two proteins had a molecular mass above 50kDa and exhibited killer activity against several B. bruxellensis strains especially in grape juice. They were active and stable at pH3.5-4.5, and temperatures between 15 and 25°C which are compatible with winemaking conditions. Furthermore, the activity of these killer toxins was not affected by the ethanol and sugar concentrations typically found in grape juice and wine. In addition, these killer toxins inhibited neither the Saccharomyces cerevisiae nor the lactic acid bacteria strains tested. These preliminary results indicated that the application of these toxins will have no effect on the main microbial agents that drive alcoholic and malolactic fermentations and further highlight the potential of using these toxins as agents to control the development of B. bruxellensis in grape juice or wine.


Assuntos
Brettanomyces/efeitos dos fármacos , Candida/química , Microbiologia de Alimentos , Micotoxinas/química , Micotoxinas/farmacologia , Vitis/microbiologia , Leveduras/metabolismo , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antifúngicos/metabolismo , Antifúngicos/farmacologia , Candida/metabolismo , Fermentação , Concentração de Íons de Hidrogênio , Lactobacillaceae/efeitos dos fármacos , Micotoxinas/isolamento & purificação , Micotoxinas/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Dióxido de Enxofre/farmacologia , Temperatura , Vinho/microbiologia
17.
Rev. Inst. Med. Trop. Säo Paulo ; 52(3): 161-162, May-June 2010.
Artigo em Inglês | LILACS | ID: lil-550348

RESUMO

The aim of this study was to report the ability of killer toxins, previously used as biotyping techniques, as a new tool to differentiate C. albicans from C. dubliniensis. The susceptibility of C. albicans and C. dubliniensis to killer toxins ranged from 33.9 to 93.3 percent and from 6.67 to 93.3 percent, respectively.


Avaliou-se a capacidade das toxinas killer, previamente utilizadas na biotipagem de C. albicans, como método para diferenciar C. albicans de C. dubliniensis. A susceptibilidade de C. albicans e C. dubliniensis às toxinas killer variou de 33,9 por cento a 93,3 por cento para C. albicans e de 6,67 por cento a 93,3 por cento para C. dubliniensis.


Assuntos
Candida/classificação , Candida/efeitos dos fármacos , Citotoxinas/farmacologia , Fatores Matadores de Levedura/farmacologia , Técnicas de Tipagem Micológica/métodos , Candida albicans/efeitos dos fármacos , Testes de Sensibilidade Microbiana
18.
Rev. argent. microbiol ; 39(4): 230-236, oct.-dic. 2007. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-634563

RESUMO

La identificación rápida y segura de los agentes etiológicos y el desarrollo de nuevos antifúngicos con blancos de acción más específicos resultarán en tratamientos de las micosis más efectivos y menos lesivos. Mediante un método molecular rápido (ITS1-5.8S ADNr-ITS2 PCR-RFLP) se identificaron 53 aislamientos de levaduras provenientes de infecciones no sistémicas registradas en hospitales públicos de la ciudad de Neuquén y en un centro oftalmológico de Buenos Aires durante el año 2005. Adicionalmente y utilizando el método de inhibición del crecimiento en placa, se evaluó la sensibilidad de estas levaduras a toxinas killer producidas por levaduras indígenas de la Patagonia y por cepas de referencia. Ocho especies de levaduras fueron identificadas entre los aislamientos clínicos: Candida albicans (52%) , Candida parapsilosis (17%) , Candida tropicalis (10%) , Candida krusei (5%) , Candida glabrata (4%) , Candida guilliermondii (4%) , Kluyveromyces lactis (4%) y Saccharomyces cerevisiae (4%) . El 69% de los aislamientos de la especie mayoritaria, C. albicans, se relacionó con infecciones vaginales. Por otra parte, el 61% de las levaduras provenientes de infecciones oculares correspondió a la especie C. parapsilosis. En las condiciones de ensayo, las toxinas producidas por las levaduras killer indígenas DVMais5 y HCMeiss5 pertenecientes a las especies Pichia anomala y P. kluyveri, respectivamente, exhibieron el mayor espectro de acción sobre las levaduras aisladas de materiales clínicos.


The use of quick and reliable yeast identification methods, as well as the development of new antifungal agents with more specific targets, will enable a more efficient treatment of mycoses. In the present work, a total of 53 clinical isolates obtained from non-systemic infections in Neuquén Hospitals and an ophthalmologic clinic in Buenos Aires during 2005, were identified by means of a rapid molecular method (ITS1-5.8S ADNr-ITS2 PCR-RFLP). Additionally, the killer susceptibility of the isolates was tested against reference and indigenous killer yeasts on plate tests. Eight yeast species were identified among the clinical isolates: Candida albicans (52%), Candida parapsilosis (17%), Candida tropicalis (10%), Candida krusei (5%), Candida glabrata (4%) , Candida guilliermondii (4%) , Kluyveromyces lactis (4%) and Saccharomyces cerevisiae (4%) . Sixty-nine percent of the isolates corresponding to the predominant species ( C. albicans) were related to vaginal infections. On the other hand, 61% of the yeasts associated with ocular infections were identified as C. parapsilosis. Two indigenous killer isolates DVMais5 and HCMeiss5, belonging to Pichia anomala and P. kluyveri respectively, exhibited the broadest killer spectrum against clinical isolates.


Assuntos
Feminino , Humanos , Masculino , Técnicas de Tipagem Micológica , Micoses/microbiologia , Micotoxinas/farmacologia , Proteínas/farmacologia , Leveduras/isolamento & purificação , Candida/efeitos dos fármacos , Candida/isolamento & purificação , Candidíase Vulvovaginal/microbiologia , Candidíase/microbiologia , Farmacorresistência Fúngica , Infecções Oculares Fúngicas/microbiologia , Fatores Matadores de Levedura , Kluyveromyces/efeitos dos fármacos , Kluyveromyces/isolamento & purificação , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/isolamento & purificação , Leveduras/efeitos dos fármacos
19.
Rev. Soc. Bras. Med. Trop ; 31(6): 523-527, nov.-dez. 1998. tab
Artigo em Português | LILACS | ID: lil-463594

RESUMO

Opportunistic infections of the oral cavity are primarily caused by Candida and frequently occur in patients with cancer who are undergoing chemotherapy and antibiotic treatment. Of the specimens received from the oral mucosa of 44 patients with cancer, 25 (56.8%) yielded Candida on culture in Sabouraud agar. Twenty four of these isolates were identified as C. albicans (96%) and 1 as C. krusei (4%). The phenotypic characteristics of these isolates showed that all of them were strongly proteolytic, had a high ability to produce phospholipase, and presented the byotypes characterized as 811 (95.8%) and 511 (4.2%) in terms of susceptibility to killer toxins.


Infecções oportunistas da cavidade bucal são primariamente causadas por fungos do gênero Candida e freqüentemente ocorrem em pacientes com câncer que estão sobtratamento quimioterápico e antibacteriano. De 44 amostras coletadas da mucosa oral de pacientes com câncer, observou-se o isolamento de 25 leveduras do gênero Candida em cultivo realizado em ágar Sabouraud-dextrose. Foram identificados Candida albicans em 24 (96%) isolados e C. krusei em 1 (4%). As características fenotípicas das amostras de Candida albicans mostraram que todos os isolados foram fortemente proteolíticos, capazes de produzir fosfolipases e possuíam os biotipos caracterizados como 811(95,8%) e 511 (4,2%) em relação a susceptibilidade às toxinas killer.


Assuntos
Humanos , Candida/isolamento & purificação , Candida/metabolismo , Candidíase Bucal/complicações , Micotoxinas/biossíntese , Mucosa Bucal/microbiologia , Neoplasias/complicações , Morte Celular , Candida albicans/enzimologia , Candida albicans/isolamento & purificação , Candida albicans/metabolismo , Candida/enzimologia , Candidíase Bucal/patologia , Endopeptidases/biossíntese , Fosfolipases/biossíntese , Testes de Sensibilidade Microbiana , Neoplasias/patologia
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