Search Results (40)

Candidozyma auris (formerly Candida auris) is an emerging multidrug-resistant pathogenic fungus with an increased ability to cause outbreaks in healthcare facilities, leading to poor patient outcomes. Since its initial discovery in 2009, C. auris has spread rapidly across continents and is now classified by both the Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO) as a critical-priority pathogen. This review summarizes current knowledge on the origin, taxonomy, microbiology, and virulence mechanisms of C. auris, emphasizing its thermotolerance, osmotolerance, and biofilm-forming capacity on biotic and abiotic surfaces, as well as aspects related to its antifungal drug resistance and management. These features, together with its genomic plasticity, contribute to persistence, transmission, and drug resistance. Emerging evidence also supports a potential link between climate change and C. auris evolution, highlighting environmental adaptation as a driver of pathogenicity. Combating C. auris will require multidisciplinary efforts to mitigate its expanding global impact. Full article

In this study, we investigated the presence and diversity of FLA in 62 environmental samples collected across Tenerife, Canary Islands, Spain including agricultural and playground soils, and on double treated water from public refrigerated fountains. Amoebae were isolated by culturing processed samples onto 2% Non-Nutrient Agar plates (NNA) which were checked daily for further processing up to molecular characterization. In this case, two approaches for molecular identification were assessed: direct multiplex qPCR targeting four potentially pathogenic FLA (Acanthamoeba spp., Vermamoeba vermiformis, Naegleria fowleri, and Balamuthia mandrillaris) DNA, and culture-based isolation followed by standard PCR and sequence analysis. Regarding qPCR results, 72.6% (45/62) of the samples were positive for at least one FLA, with V. vermiformis (37/62) and Acanthamoeba spp. (34/62) being the most frequent. Moreover, B. mandrillaris was detected for the first time in the Canary Islands in 6 out of 62 samples. Results from standard PCR from cultured isolates confirmed the presence of Acanthamoeba (mainly genotype T4) and Vermamoeba and also allowed the identification of Vahlkampfia and Vannella genera, as well as the genus Rhogostoma—its first report in the Canary Islands. Thermotolerance and osmotolerance assays were performed on Acanthamoeba spp. and, innovatively, on V. vermiformis isolates. Both were capable of surviving at 37 °C and during incubation with 0.5 M mannitol, suggesting potential pathogenicity. However, growth was significantly impaired under harsher conditions (42 °C and 1 M mannitol). These findings underscore the widespread occurrence of FLA in public and agricultural environments in Tenerife and highlight their potential risk to public health. Their ability to act as carriers of pathogenic bacteria/viruses further reinforces the need for routine surveillance and preventive measures in the environment. Full article

Heterofermentative lactic acid bacteria (LABhe) can help to increase aerobic stability of silages. As high dry matter silages are especially prone to aerobic deterioration, the question was whether an in vitro test could help to select for LABhe with tolerance to these conditions, that is, to high osmolality. A medium proven for homofermentative lactic acid bacteria (LABho) with high sugar concentration was applied in vitro while validating the results in high DM grass silages in situ. The pH was measured at 0, 24, 36 and 48 h or after 5 d as indicator for the bacterial fermentation activity and compared. As a result, LABhe showed to be far less susceptible to high DM and osmolality respectively, compared to LABho. It was concluded that the in vitro test allows for qualitative assessment of the osmotolerance of LABhe, but not for quantitative ranking between LABhe strains. Full article

Candidozyma auris is a multidrug-resistant, thermo- and osmotolerant yeast capable of persisting on biotic and abiotic surfaces, attributes likely linked to its cell wall composition. Here, seven putative genes encoding yapsins, aspartyl proteases GPI-anchored to the membrane or cell wall, were identified in the genomes of C. auris CJ97 and 20-1498, from clades III and IV, respectively. The C. auris YPS1 gene is orthologous to the SAP9 of C. albicans. The YPS7 gene is orthologous to YPS7 in C. glabrata and S. cerevisiae, so that they may share similar roles. An in silico analysis suggested an interaction between pepstatin and the catalytic domain of Yps1 and Yps7. Although this inhibitor, when combined with caffeine, had a subtle effect on the growth of C. auris, it induced alterations in the cell wall. CauYPS1 and CauYPS7 expression increased under nutrient starvation and NaCl, and at 42 °C. The transcriptome of the 20-1498 strain suggests that autophagy may play a role in thermal stress, probably degrading deleterious proteins or maintaining cell wall and vacuolar homeostasis. Therefore, CauYps1 and CauYps7 may play a role in the cell wall integrity of C. auris in stress conditions, and they could be a target of new antifungal or antivirulence agents. Full article

To learn about the yeast biodiversity of Hungarian honeys and to isolate osmotolerant yeasts, fifteen different honey varieties, beeswax, and bee bread were purchased, and samples of another, but highly osmotic material, tree sap (cherry, sour cherry, and plum), were collected from the northeastern region of the country. In total, 60 yeast strains were isolated and their taxonomic positions were determined by barcode sequences using ITS1-NL4 primers. The honey products contained mostly Zygosaccharomyces and Starmerella species. In addition, Hanseniaspora uvarum, Rhodotorula mucilaginosa and diobovata, Sporobolomyces roseus, Filobasidium magnum, Naganishia sp., and Aureobasidium pullulans were also present in smaller numbers. In contrast, tree saps contained Metschnikowia and Pichia fermentas cells. Further results suggest that some of the yeasts in honey can only “survive”, while others can propagate at high sugar levels, generally between 600 and 700 mg/g, with a predominance of fructose. Properties important for pathogenicity, such as invasive hyphae production, gelatin melting ability, and growth at 37 °C, were also examined. Hanseniaspora uvarum and Pichia fermentans representatives seemed to be negative for gelatin hydrolysis, while the other strains were able to melt gelatin. Although some of the strains could produce hyphae-like structures at 25 °C, none of them could grow at 37 °C. Full article

The traditional marination process enhances food flavor and inhibits microbial growth. However, in hyperosmotic environments, microorganisms can activate stress responses to ensure survival, potentially compromising food safety. This study investigated the osmotolerance mechanisms of Salmonella Derby (S. Derby) by comparing a wild-type strain (S. D-WT) and an osmotolerant strain (S. D-OT) under NaCl-induced hyperosmotic stress. Both strains were subjected to 0.85%, 4%, and 16% NaCl for 0, 8, and 16 days, and their growth behavior, membrane integrity, intracellular osmoprotectant content, and transcription of related genes were evaluated. By day 16, both strains showed a growth delay of approximately 3 h. S. D-OT maintained better membrane integrity and exhibited higher intracellular levels of osmoprotectants (K⁺, trehalose, and proline), which aligned with the upregulation of the transcriptional levels of kdpC, kuP, rpoS, and proU. These findings indicated that S. D-OT achieved improved osmotic stress tolerance by regulating osmoprotectant synthesis and maintaining intracellular homeostasis. In contrast, S. D-WT displayed greater resistance to multiple antibiotics (gentamicin, ciprofloxacin, trimethoprim-sulfamethoxazole, and chloramphenicol) under 4% and 16% NaCl conditions, which may pose a higher food safety risk. Overall, this study provides insights for improving microbial control strategies in preserved foods and mitigating foodborne disease risks. Full article

Smear-ripened cheeses are distinguished by their complex microbiota, which play an important role in ripening, flavour development, texture and microbiological safety. Although commercial production typically relies on defined starter and adjunct cultures, microorganisms from the production environment may also shape the product’s characteristics. This study examined the microbial composition of smear-ripened cheeses from six commercial manufacturers using culture and culture-independent techniques, including 16S rRNA gene sequencing and sequencing of internal transcribed spacers. A limited number of microorganisms was recovered by culture, with 37 different isolates identified across all samples. Sequencing of the 16S rRNA gene and internal transcribed spacers revealed 75 and 7 distinct operational taxonomic units, respectively. The microbiota composition reflected the contribution of both mesophilic and thermophilic starter and adjunct cultures, alongside microorganisms originating from the production environment. These included various psychrotrophic bacteria, marine (i.e., osmotolerant) bacteria, and other halophiles from Proteobacteria (Psychrobacter, Pseudoalteromonas, Marinomonas, and Vibrio), Firmicutes (Vagococcus and Marinilactibacillus), Actinobacteriota (Glutamicibacter), Bacteroidota (Winogradskyella and Brumimicrobium), Campylobacterota (Malaciobacter) and Fusobacteriota (Psychrilyobacter) specific to the environment of particular manufacturers. The results indicate that, although pasteurised milk and defined starter cultures are used in commercial production, microorganisms originating from the cheese factory environment form a substantial part of the microbiota of smear-ripened cheese. Full article

Droughts are projected to become prevalent throughout the 21st century, endangering agricultural productivity and global food security. To address these challenges, novel strategies to enhance water management and augment plant resilience are imperative. Bacterial encapsulation has emerged as a promising approach, offering benefits such as enhanced bacterial survival, soil compatibility, and sustainable plant growth. This study evaluated the osmotolerance of bacteria from arid environments and determined their plant growth-promoting ability in drought conditions. The encapsulation of these bacteria in bio-compatible capsules led to a substantial enhancement in the performance of maize plants under drought stress. Maize plants treated with encapsulated bacteria demonstrated a 35% increase in root biomass and a 28% enhancement in shoot growth compared to untreated controls. Furthermore, significant physiological and biochemical adaptations were observed, including a 45% increase in photosynthetic pigment concentration and higher osmolyte levels, which contributed to improved drought stress tolerance. The findings of this study demonstrate the potential of encapsulated bacteria to enhance maize resilience to drought, thereby supporting robust growth under water-limited conditions. This approach presents a sustainable strategy to improve drought tolerance, and it may reduce irrigation dependency and maintain crop yields in the face of increasing climate uncertainty. Full article

This study aimed to address the need for effective probiotics and antibacterial agents to combat American foulbrood disease in honeybees, caused by Paenibacillus larvae. In the context of declining honeybee populations due to pathogens, we isolated eight lactic acid bacteria (LAB) strains from honeybee larvae (Apis mellifera L.) and evaluated their probiotic potential and inhibitory effects against P. larvae. Methods included probiotic property assessments, such as acid and bile salt resistance, hydrophobicity, auto-aggregation, co-aggregation with P. larvae, antioxidant capacities, osmotolerance to 50% sucrose, and antibiotic susceptibility. Results indicated that the GL3 strain exhibited superior probiotic attributes and potent inhibitory effects on P. larvae. Whole-genome sequencing revealed GL3 to be an Enterococcus lactis strain with genetic features tailored to the honeybee larval gut environment. Pangenome analysis highlighted genetic diversity among E. lactis strains, while molecular docking analysis identified aborycin, a lasso peptide produced by GL3, as a promising inhibitor of bacterial cell wall synthesis. These findings suggested that GL3 was a promising probiotic candidate and antibacterial agent for honeybee health management, warranting further investigation into its in vivo efficacy and potential applications in beekeeping practices. Full article

The Namib Desert is characterized by a number of abiotic stresses, including high temperature, high salinity, osmotic pressure, alkaline pH, and limited water availability. In such environments, dry soils typically exhibit a low water potential, scarce nutrients, and high concentrations of dissolved ions, collectively creating a challenging habitat for microbial life. In this study, 89 bacterial isolates belonging to 20 genera were identified. Bacteria demonstrated significant osmotolerance, with some strains thriving at polyethylene glycol (PEG) concentrations exceeding 20%. Furthermore, these bacteria demonstrated halotolerance, high pH tolerance, and capacity to produce plant growth-promoting (PGP) traits under conditions of osmotic stress. Osmotolerant bacteria exhibited higher proficiency in siderophore production, potassium solubilization, and phosphorus solubilization, all of which are critical for supporting plant growth in nutrient-scarce and stressful environments, such as deserts. However, alginate production was higher in isolates that were less osmotolerant, indicating the potential for a compensatory mechanism in strains that were more sensitive. These findings highlight the complex strategies employed by desert bacteria to survive and support host plants in extreme environments. The present study not only enhances our understanding of microbial adaptations in arid ecosystems, but also provides important information for the development of potential applications for these bacteria in the reclamation of arid land and agricultural practices aimed at improving crop resilience to abiotic stress. Full article

This review explores the production of the fungal polysaccharide pullulan by mutants and natural isolates of Aureobasidium species using strain improvement. Pullulan is a neutral polysaccharide gum whose structure is a maltotriose-containing glucan. This polysaccharide gum has applications in the fields of food, pharmaceuticals, biomedical and wastewater treatment. The strain improvement of Aureobasidium species has focused on the pullulan production process, including the isolation of strains exhibiting reduced pigmentation, polysaccharide overproduction, the production of pullulan with variable molecular weight, and increased osmotolerant strains promoting pullulan production at high carbon source concentrations and pullulan production on hemicellulosic substrates. The majority of studies have emphasized the isolation of reduced pigmentation and pullulan hyperproducer strains since the goal of large-scale commercial pullulan production is to synthesize non-pigmented polysaccharides. A promising area of strain improvement is the isolation of strains that synthesize authentic pullulan from hemicellulosic substrates. If strain improvement in this area is successful, the goal of commercially producing pullulan at a competitive cost will eventually be achieved. Full article

The Tajogaite Volcano erupted on the western slope of the Cumbre Vieja mountain range on La Palma Island in the Canary Islands, Spain, in 2021. As one of the multiple consequences of this eruption, a layer of tephra was deposited, to a variable extent, over a large part of the island. Tephra deposits affect all aspects of vegetation recovery, the water cycle, and the long-term availability of volcanic nutrients. Protozoa, including free-living amoeba (FLA), are known to be among the first microorganisms capable of colonizing harsh environments. In the present study, the presence of FLA has been evaluated in the Tajogaite Volcano deposits. Samples of the tephra were collected and incubated at 26 °C on 2% non-nutrient agar plates with a layer of heat-killed E. coli. Morphological features, as well as the DF3 region sequence of the 18S rDNA, confirmed the presence of a T4 genotype strain of Acanthamoeba. Thermotolerance and osmotolerance assays were used to evaluate the strain’s pathogenic potential. This strain was considered thermotolerant but poorly osmotolerant. To the best of our knowledge, this is the first report of Acanthamoeba being isolated from a recently erupted volcano. Full article

Coniella vitis is a dominant phytopathogen of grape white rot in China, significantly impacting grape yield and quality. Previous studies showed that the growth and pathogenicity of C. vitis were affected by the environmental pH. Arrestin-like protein PalF plays a key role in mediating the activation of an intracellular-signaling cascade in response to alkaline ambient. However, it remains unclear whether palF affects the growth, development, and virulence of C. vitis during the sensing of environmental pH changes. In this study, we identified a homologous gene of PalF/Rim8 in C. vitis and constructed CvpalF-silenced strains via RNA interference. CvpalF-silenced strains exhibited impaired fungal growth at neutral/alkaline pH, accompanied by reduced pathogenicity compared to the wild-type (WT) and empty vector control (CK) strains. The distance between the hyphal branches was significantly increased in the CvpalF-silenced strains. Additionally, CvpalF-silenced strains showed increased sensitivity to NaCl, H₂O₂, and Congo red, and decreased sensitive to CaSO₄. RT-qPCR analysis demonstrated that the expression level of genes related to pectinase and cellulase were significantly down-regulated in CvpalF-silenced strains compared to WT and CK strains. Moreover, the expression of PacC, PalA/B/C/F/H/I was directly or indirectly affected by silencing CvpalF. Additionally, the expression of genes related to plant cell wall-degrading enzymes, which are key virulence factors for plant pathogenic fungi, was regulated by CvpalF. Our results indicate the important roles of CvpalF in growth, osmotolerance, and pathogenicity in C. vitis. Full article

Lactic acid bacteria (LAB) should not only survive, but also perform under increased osmotic pressure in the process of ensiling, which results from the best practice of wilting forage. Simple laboratory protocols are needed to select suitable LAB strains as inoculants for high dry matter (DM) conditions. The aim of this study was to simulate conditions of high osmolality without inducing salt stress and to select a suitable indicator of LAB performance. For that, an MRS medium was enriched with increasing concentrations of glucose and fructose plus a maximum of 28 g KCl/L until achieving an osmolality of 2.4 osmol/kg. Both, growth in the inoculated medium and pH decline, were then compared to the LAB performance in the basic medium. The latter was clearly delayed in the new medium. Finally, the method was validated by comparing the pH of small-scale grass silages of 30–35 and 45–49% target DM after 3–5 days of ensiling to the pH values of the microbiological growth medium. The pH levels of treatments with the homofermentative LAB were clearly attributable to the dry matter or the sugar concentration, respectively. The developed liquid growth medium sufficiently approximates high DM conditions to select for the osmotolerant homofermentative LAB. Full article

This study was conducted to investigate the genotypic and phenotypic characteristics of lactic acid bacteria (LAB) associated with forage plants in the native grassland of western Inner Mongolia and to evaluate their effects on alfalfa silage fermentation. Forage plants and their spontaneous fermentation silages were analysed using culture-based techniques for LAB isolation; the phenotypic properties and 16S rDNA and pheS or rpoA gene sequences of the isolates were evaluated; alfalfa was ensiled with four additive combinations: Lactiplantibacillus plantarum subsp. plantarum (GI19), Lact. plantarum subsp. plantarum and Pediococcus pentosaceus (GI19+GI51), GI19 and 20 g/kg fresh matter of sucrose (GI19+S), and GI19+GI51+S, for 60 d. A total of 73 strains belonging to 16 species were isolated. All isolates grew at 5–45 °C and in 3.0% NaCl, and most of them grew in 6.5% NaCl. Enterococcus faecalis and Lact. plantarum were 26.03% and 17.81% of the total isolates, respectively. All additives improved the silage quality, while GI19+S was more effective for alfalfa ensiling with a higher lactic acid content and lower pH, undesirable microorganism counts, and acetic acid and NH₃-N contents than remnant additives. In conclusion, the LAB species were diverse, and most of them possessed good cryotolerance and osmotolerance; GI19+S was the optimal inoculant for alfalfa fermentation improvement. Full article