Chemical composition, antifungal, and anti-virulence action of the stem bark of Hancornia speciosa Gomes (Apocynaceae) against Candida spp.

ETHNOPHARMACOLOGICAL RELEVANCE: Hancornia speciosa Gomes is a fruit and medicinal species used for treating infectious diseases of the genitourinary system. However, its mechanism of action against microbes is still not fully understood. Infections in the genitourinary system caused by Candida spp. are associated with its fungal resistance and pathogenicity. New plant-derived compounds are an alternative to fight these Candida infections. AIM OF THE STUDY: The objective of this study was to evaluate the anti-Candida effects of extracts of the stem bark of H. speciosa. This research investigated the chemical composition of sulfuric ether (EEHS) and methanolic (MEHS) extracts, their drug-modifying action on fluconazole, and their anti-virulence action on the morphological transition of Candida species. MATERIALS AND METHODS: The extracts (EEHS and MEHS) of the stem bark of H. speciosa were chemically characterized via qualitative phytochemical screening and by liquid chromatography coupled with mass spectrometry (UPLC-MS-ESI-QTOF). The extracts were evaluated regarding their antifungal effects and fluconazole-modifying activity against Candida albicans, Candida krusei, and Candida tropicalis using the broth microdilution method. Additionally, the study evaluated the inhibition of fungal virulence in Candida species through morphological transition assays. RESULTS: The phytochemical screening revealed the presence of anthocyanidins, anthocyanins, aurones, catechins, chalcones, flavones, flavonols, flavanones, leucoanthocyanidins, tannins (condensed and pyrogallic), and xanthones in both extracts of the stem bark of H. speciosa. The UPLC-MS-ESI-QTOF analysis identified the same compounds in both extracts, predominating phenolic compounds. Some compounds were first time recorded in this species: gluconic acid, cinchonain IIb, cinchonain Ib isomer, and lariciresinol hexoside isomers. Most of the intrinsic antifungal activity was observed for the MEHS against C. krusei (IC50: 58.41 µg/mL). At subinhibitory concentrations (MC/8), the EEHS enhanced the action of fluconazole against all Candida strains. The MEHS exhibited greater efficacy than fluconazole inhibiting C. krusei growth. The EEHS completely inhibited hyphae appearance and reduced pseudohyphae formation in C. albicans. CONCLUSION: The stem bark of H. speciosa is a rich source of bioactive compounds, especially phenolic. Phenolic compounds can have important roles in fighting infectious diseases of the genitourinary system, such as candidiasis. The extracts of H. speciosa improved the action of the drug fluconazole against Candida species, inhibited hyphae appearance, and reduced pseudohyphae formation. The results of this study can support the development of new therapeutics against resistant strains of Candida.

Determination of microbial numbers in anaerobically digested biofertilisers.

This study aimed to quantity total numbers of bacteria, fungi and archaea in different types of commercial liquid anaerobic digestates, and to identify common patterns in their microbial numbers post-digestion and possible implications of their use as biofertiliser. Relationships between microbial numbers and physical-chemical traits of the digestates were also investigated. Quantification was performed using culturable and molecular (quantitative PCR) approaches. Bacterial and fungal CFUs ranged up to five orders of magnitude (105-1010; 0-105 g-1 DW, respectively) between different types of anaerobic digestates. Bacterial, archaeal and fungal gene copy numbers (GCN) varied by two orders of magnitude (108-1010; 107-109; 104-106 g-1 DW, respectively) between digestates. All microbial variables analysed showed significant differences between the different types of anaerobic digestate investigated (p < 0.05). Culturable microbial numbers for fungi (6.43 × 104 CFU g-1 DW) were much lower than for bacteria (2.23 × 109 CFU g-1 DW). Gene copy numbers were highest for bacteria (16S) (1.09 × 1010 g-1 DW), followed by archaea (16S) (5.87 × 108 g-1 DW), and fungi (18S) (1.77 × 106 g-1 DW). Liquid anaerobic digestates were predominantly dominated by bacteria, followed by archaeal and fungal populations. At 50% similarity level, the microbial profiles of the eleven anaerobic digestates tested separated into just two groups, indicating a broad relative degree of similarity in terms of microbial numbers. Higher bacterial (16S) GCN was associated with low OM and C/N ratio in digestates.

Physical-chemical traits, phytotoxicity and pathogen detection in liquid anaerobic digestates.

Anaerobic digestates, which are co-products from biogas production, have been recognised as potential biofertilisers for their benefits in nutrient recovery and recycling of different types of organic wastes. Due to the increasing number of different types of organic wastes being used to produce biogas, it is necessary to identify how different types of anaerobic digestates vary in their physical-chemical traits, and how these can impact upon their use as fertilisers. In addition, safe land spreading of anaerobic digestates must be within recommended limits for potentially toxic elements (PTEs) and pathogens. This study analysed physical-chemical traits, phytotoxicity, PTEs and indicator pathogens in a set of eleven different commercial liquid anaerobic digestates from Ireland and the UK, and compared them to the Irish draft standard for digestate. Liquid anaerobic digestates exhibited significant differences (P < 0.001) for most of the physical and chemical traits evaluated, with higher variability found for dry matter (DM) and K (CV = 17.2 and 16.8 respectively), and lower variation for pH and P (CV = 1.78 and 3.55 respectively). PTE concentrations were in general within recommended limits; nevertheless, some digestates showed higher concentrations than the recommended limits for Pb, Zn and Cu. Digestate from wastewater treatment feedstock was shown to be high in PTEs. Anaerobic digestates were found to negatively affect early stages of seed germination, but phytotoxicity effects were decreased by dilution in water. Levels of Salmonella spp. and E. coli were within recommended limits for most of the anaerobic digestates analysed.