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Abstract Polyethylene (PE) is high molecular weight synthetic polymer, very hydrofobic and hardly biodegradable. To increase polyethylene bio-degradability it is very important to find microorganisms that improve the PE hydrophilic level and/or reduce the length of its polymeric chain by oxidation. In this study, we isolated Cladosporium halotolerans, a fungal species, from the gastric system of Galleria mellonella larvae. Here, we show that C. halotolerans grows in the presence of PE polymer, it is able to interact with plastic material through its hyphae and secretes enzymes involved in PE degradation. In this article, a fungus is isolated from the gastrointestinal tract of the G. mellonella larva, which infests hives by feeding on wax. The similarity of structure between the wax and the polyethylene makes the microorganism suitable for the degradation of plastic materials that are difficult to eliminate.

Microbial volatiles have beneficial roles in the agricultural ecological system, enhancing plant growth and inducing systemic resistance against plant pathogens without being hazardous to the environment. The interactions of plant and fungal volatiles have been extensively studied, but there is limited research specifically elucidating the effects of distinct volatile organic compounds (VOCs) on plant growth promotion. The current study was conducted to investigate the impact of VOCs from Cladosporium halotolerans NGPF1 on plant growth, and to elucidate the mechanisms for the plant growth-promoting (PGP) activity of these VOCs. The VOCs from C. halotolerans NGPF1 significantly promoted plant growth compared with the control, and this PGP activity of the VOCs was culture medium-dependent. Headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography–mass spectrometry (GC–MS) identified two VOC structures with profiles that differed depending on the culture medium. The two compounds that were only produced in potato dextrose (PD) medium were identified as 2-methyl-butanal and 3-methyl-butanal, and both modulated plant growth promotion and root system development. The PGP effects of the identified synthetic compounds were analyzed individually and in blends using N. benthamiana plants. A blend of the two VOCs enhanced growth promotion and root system development compared with the individual compounds. Furthermore, real-time PCR revealed markedly increased expression of genes involved in auxin, expansin, and gibberellin biosynthesis and metabolism in plant leaves exposed to the two volatile blends, while cytokinin and ethylene expression levels were decreased or similar in comparison with the control. These findings demonstrate that naturally occurring fungal VOCs can induce plant growth promotion and provide new insights into the mechanism of PGP activity. The application of stimulatory volatiles for growth enhancement could be used in the agricultural industry to increase crop yield.

Abstract Halotolerant spoilage fungi, represented by Cladosporium halotolerans, make it difficult to ensure the microbiological stability of herbal beverages, such as chrysanthemum tea, and have renewed interest in natural antifungal options. In this work, a spoilage strain of C. halotolerans was isolated from a commercial chrysanthemum tea product and used as an indicator organism to assess the antifungal activity of eight plant-derived extracts. Plum extract and citral showed the most pronounced growth inhibition and were, therefore, incorporated into chrysanthemum tea to form composite preservative systems. Both a ternary formulation containing plum extract, citral, and ginger extract, and a simpler binary plum–citral formulation, markedly reduced C. halotolerans growth in chrysanthemum tea, with the binary system ultimately selected because it combined strong antifungal efficacy with better flavour compatibility. Scanning electron microscopy revealed the obvious deformation and collapse of C. halotolerans cells after exposure to the plum–citral composite, suggesting serious damage to the cell envelope. RNA-seq profiling further showed widespread changes in gene expression in response to the composite, and Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment analyses indicated that many differentially expressed genes were involved in central metabolic pathways, and organic-acid- and small-molecule–related metabolic processes. Taken together, these results indicate that a chrysanthemum tea-based composite of plum extract and citral is a promising natural antifungal system against C. halotolerans, acting mainly through the disruption of the cell envelope and the disturbance of key metabolic functions. Graphical Abstract Graphical Abstract

This study explored the great potential of endophytic fungi associated with red seaweed Gracilaria sp. and brown seaweed Sargassum sp. of the Bay of Bengal, Bangladesh, for the first time. Endophytic fungi were identified taxonomically by morphological features and molecular characterisation (ITS sequence). The identification of six fungal isolates revealed five different fungal species belonging to four genera, namely, Aspergillus subversicolor , A. terreus and Cladosporium halotolerans isolated from Gracilaria sp. and Chaetomium globosum , A. terreus and Curvularia perotidis isolated from Sargassum sp. The ethyl acetate extracts of fungal endophytes were evaluated for antimicrobial activity, DPPH radical scavenging activity, and brine shrimp lethality bioassay. Amongst all the fungal extracts evaluated in this study, four showed mild to moderate inhibitory activity (10–14 mm) against the tested bacterial strains. Exceptionally, Chaetomium globosum exerted significant antibacterial activity with the highest zone of inhibition (21 ± 0.3 mm) against the Gram-negative bacterial strain Pseudomonas aeruginosa and also showed moderate antifungal activity (13 ± 0.9 mm) against A. niger . Sargassum sp.-derived A. terreus exhibited the strongest DPPH radical scavenging activity (IC 50 value of 7.88 ± 0.09 µg/mL). All the fungal extracts have significant lethality against brine shrimp nauplii (LC 50 value of ≤ 20.39 ± 4.04 µg/mL), while Curvularia perotidis and Cladosporium halotolerans were the most effective (LC 50 values of 9.30 ± 2.96 µg/mL and 9.94 ± 3.49 µg/mL, respectively). Gas chromatography-mass spectrometry (GC-MS) analysis of the crude extracts identified the presence of several chemical compounds. These bioactive chemical constituents might contribute to the exhibited bioactivity in this study. The current study’s findings support the beneficial impacts of the fungal endophytes on exerting biological activities and consequently as valuable resources of bioactive compounds.

Background Cladosporium halotolerans is a saprobic fungus, rarely implicated in human infections. The identification is challenging due to non-specific phenotypic features. Objective To decipher clinical spectrum, microbiological and susceptibility profile of clinical and environmental isolates of Cladosporium halotolerans . Method All the isolates identified as Cladosporium halotolerans deposited in National Culture Collection for Pathogenic Fungi (NCCPF), Postgraduate Institute of Medical Education and Research, Chandigarh, India were revived. Phenotypic and molecular characterization targeting internal transcribed spacer (ITS) region of ribosomal DNA, large subunit of ribosomal DNA (LSU; NL1 and NL4), actin ( ACT ) and beta-tubulin ( TUB ) was done. Scanning electron microscopy (SEM) was performed to determine any phenotypic variations. Antifungal susceptibility testing (AFST) was carried out for eight antifungal agents as per CLSI M38 Ed3 guidelines. We also performed systematic literature review of all the cases of Cladosporium halotolerans reported till date. Results A total of four isolates (clinical, n = 3; soil, n = 1) identified as Cladosporium halotolerans were included in the study. The clinical sites were skin, maxillary tissue and nail. All patients were apparently immunocompetent, and history of trauma was recorded in one patient. All patients improved on antifungal therapy. The cultures revealed growth of black mycelial fungus and microscopic examination demonstrated dematiaceous septate hyphae with erect conidiophores and conidia in branched acropetal chains. Based on molecular methods, all the four isolates were identified as C. halotolerans. SEM revealed no variation in length and width of the conidia, conidiophores, ramoconidium and hyphae among the isolates. All molecular targets, such as ITS region, LSU (partially sequenced), ACT and TUB were able to differentiate the isolates. Minimum inhibitory concentrations for antifungals were: triazoles (0.12–2 μg/ml), amphotericin B (4 μg/ml) and echinocandins (2–8 μg/ml). Conclusion We report role of the rarely isolated dematiaceous fungus, C. halotolerans, in causing human infections. The study emphasizes the role of molecular methods in precisely identifying these species. Triazoles are more active against these black fungi compared to polyenes or echinocandins.

The effect of microbial activity on the deterioration of archaeological stone items is a worldwide issue, and conserving them with low-toxicity, ecologically benign and naturally biocides is a difficult undertaking. Molecular identification of the microbial communities from a deteriorated archaeological object (Ptahshepses stone statue) located from the Saqqara excavation, Giza, Egypt was recorded. Six essential oils (EOs) (black cumin, clove, geranium, lavender, lemongrass, and thyme) were tested for antimicrobial activity against six degrading fungal and bacterial species. Alternaria alternate, Aspergillus flavus, A. niger, Cladosporium halotolerans, Penicillium crustosum, and Trichoderma viride and three bacterial species, Pseudomonas protegens, P. putida, and Serratia odorifera, were isolated. Of the most effective EOs, thyme showed the highest inhibition percentage (143.4%) against Serratia odorifera, followed by P. putida (135%), and Pseudomonas protegens (131.5%). Lemongrass and clove EOs had minimum inhibitory concentrations (MICs) ranging from 0.5 µL/mL to 2 µL/mL for all isolated deteriorated fungal and bacterial species, while the lowest efficiency EOs were lavender, geranium and black cumin. It can be concluded that thyme and lemongrass EOs have a potential use for protecting the Ptahshepses stone statue from microbial deterioration.

This report presents the pathologic findings associated with disseminated infection due to Cladosporium halotolerans in a dog that was simultaneously infected with canine adenovirus-1 (CAdV-1) and canine parvovirus-2 (CPV-2). A 12-year-old, mixed breed dog, with a clinical history of neurological manifestations was submitted for routine autopsy due to poor prognosis. The principal pathologic findings were mycotic necrotizing nephritis, hepatitis, and splenitis with embolic dissemination to the brain resulting in mycotic necrotizing meningoencephalitis, ventriculitis, choroid plexitis, and obstructive hydrocephalus associated with intralesional and intravascular septate pigmented fungi. PCR and sequencing of the ITS region of fungi revealed that the intralesional fungal organisms had 82% nucleotide identity with members of the Cladosporium sphaerospermum complex of organisms. However, a PCR assay and sequencing of the beta tubulin gene confirmed that the organism identified in this dog had 100% nucleotide sequence identity with C. halotolerans . Using immunohistochemistry, intralesional antigens of CAdV-1 were identified within the epithelial cells of the liver and lungs; there was positive immunolabeling for CPV-2 antigens in degenerated cardiomyocytes. These findings confirmed the active participation of C. halotolerans in the development of disseminated cladosporiosis in this dog and represent a rare occurrence of concomitant infection with CAdV-1 and CPV-2.

Salinization induced by salt stress is a critical environmental factor, and limits the expansion of agricultural areas and population distribution in continental regions, including Saudi Arabia. Common beans, a vital source of protein, energy, and dietary fibers, are negatively affected by salt stress. In this research, the endophytic fungus Cladosporium halotolerans was utilized to remediate saline soils and enhance common bean growth and resilience. The results of this study demonstrated that soil treatment with C. halotolerans enhances the soil properties by decreasing soil pH and increasing soil organic matter content under saline conditions. Inoculation by C. halotolerans also significantly improved plant growth parameters, induced systemic resistance to salinity, and increased the levels of chlorophyll b and carotenoids. Fungal inoculation also causes stress relief as indicated by reducing malondialdehyde concentration by 27.4% lower than stressed plants. Microscopic images revealed the active association and colonization of C. halotolerans within the roots of the Phaseolus vulgaris both under control and saline conditions. Therefore, utilizing endophytic fungi C. halotolerans for saline soil remediation appears to be a promising alternative in plant treatments, highlighting their potential as valuable resources for both research and commercial applications under salinity stress.