Investigation of culturable human gut mycobiota from the segamat community in Johor, Malaysia.

Although several studies have already been carried out in investigating the general profile of the gut mycobiome across several countries, there has yet to be an officially established baseline of a healthy human gut mycobiome, to the best of our knowledge. Microbial composition within the gastrointestinal tract differ across individuals worldwide, and most human gut fungi studies concentrate specifically on individuals from developed countries or diseased cohorts. The present study is the first culture-dependent community study assessing the prevalence and diversity of gut fungi among different ethnic groups from South East Asia. Samples were obtained from a multi-ethnic semi-rural community from Segamat in southern Malaysia. Faecal samples were screened for culturable fungi and questionnaire data analysis was performed. Culturable fungi were present in 45% of the participants' stool samples. Ethnicity had an impact on fungal prevalence and density in stool samples. The prevalence of resistance to fluconazole, itraconazole, voriconazole and 5-fluorocytosine, from the Segamat community, were 14%, 14%, 11% and 7% respectively. It was found that Jakun individuals had lower levels of antifungal resistance irrespective of the drug tested, and male participants had more fluconazole resistant yeast in their stool samples. Two novel point mutations were identified in the ERG11 gene from one azole resistant Candida glabrata, suggesting a possible cause of the occurrence of antifungal resistant isolates in the participant's faecal sample.

Lignin-cellulose complexes derived from agricultural wastes for combined antibacterial and tissue engineering scaffolds for cutaneous leishmaniasis wounds.

Bacterial infections in wounds significantly impair the healing process. The use of natural antibacterial products over synthetic antibiotics has emerged as a new trend to address antimicrobial resistance. An ideal tissue engineering scaffold to treat infected wounds should possess antibacterial properties, while simultaneously promoting tissue regrowth. Synthesis of hydrogel scaffolds with antibacterial properties using hemp shive (HT1/HT2) lignin, sugarcane bagasse (SCB) lignin and cellulose was carried out. All lignin samples had low molecular weights and were constituted of G-type ß-5 dimers, linked by ß-O-4 bonds, as determined by MALDI-TOF-MS. Hemp lignin was more cytotoxic to mouse fibroblasts (L929) compared to SCB lignin. All lignin samples demonstrated antibacterial properties against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis, with greater efficiency against Gram-negative strains. 3D hydrogels were engineered by crosslinking SCB lignin with SCB cellulose in varying weight ratios in the presence of epichlorohydrin. The stiffness of the hydrogels could be tailored by varying the lignin concentration. All hydrogels were biocompatible; however, better fibroblast adhesion was observed on the blended hydrogels compared to the 100% cellulose hydrogel, with the cellulose : lignin 70 : 30 hydrogel showing the highest L929 proliferation and best antibacterial properties with a 24-hour bacterial growth reduction ranging from 30.8 to 57.3%.

Polysucrose hydrogel loaded with natural molecules/extracts for multiphase-directed sustainable wound healing.

Natural molecules/extracts have numerous beneficial effects on wound healing processes which are challenged by appropriate use and non-toxic dosage. Polysucrose-based (PSucMA) hydrogels have been synthesized with in situ loading of one or more natural molecules/extracts namely Manuka honey (MH), Eucalyptus honey (EH1, EH2), Ginkgo biloba (GK), thymol (THY) and metformin (MET). EH1 presented low amounts of hydroxymethylfurfural and methylglyoxal compared to MH indicating that EH1 was not temperature-abused. It also showed high diastase activity and conductivity. GK was added to PSucMA solution along with other additives including MH, EH1 and MET and crosslinked to form dual loaded hydrogels. The in vitro release profiles of EH1, MH, GK and THY from the hydrogels followed the exponential Korsmeyer-Peppas equation, with a release exponent value of less than 0.5 indicating a quasi-Fickian diffusion mechanism. The IC50 values of these natural products using L929 fibroblasts and RAW 264.7 macrophages indicated that EH1, MH and GK were cytocompatible at relatively high concentrations compared to MET, THY and curcumin used as a control. MH and EH1 induced high IL6 concentration compared to GK. In vitro studies were modelled to mimic the overlapping wound healing phases using human dermal fibroblasts (HDFs), macrophages and human umbilical endothelial cells (HUVECs) in dual culture. HDFs showed a highly interconnected cellular network on GK loaded scaffolds. EH1 loaded scaffolds were seen to induce formation of spheroids which increased in number and size in co-culture studies. The SEM images of HDF/HUVEC seeded GK, GKMH and GKEH1 loaded hydrogels indicated formation of vacuoles and lumen structures. These results indicated that a combination of GK and EH1 in the hydrogel scaffold would accelerate tissue regeneration by acting on the four overlapping phases of wound healing.

Molecular characterization and comparative genomic analysis of Acinetobacter baumannii isolated from the community and the hospital: an epidemiological study in Segamat, Malaysia.

Acinetobacter baumannii is a common cause of multidrug-resistant (MDR) nosocomial infections around the world. However, little is known about the persistence and dynamics of A. baumannii in a healthy community. This study investigated the role of the community as a prospective reservoir for A. baumannii and explored possible links between hospital and community isolates. A total of 12 independent A. baumannii strains were isolated from human faecal samples from the community in Segamat, Malaysia, in 2018 and 2019. Another 15 were obtained in 2020 from patients at the co-located tertiary public hospital. The antimicrobial resistance profile and biofilm formation ability were analysed, and the relatedness of community and hospital isolates was determined using whole-genome sequencing (WGS). Antibiotic profile analysis revealed that 12 out of 15 hospital isolates were MDR, but none of the community isolates were MDR. However, phylogenetic analysis based on single-nucleotide polymorphisms (SNPs) and a pangenome analysis of core genes showed clustering between four community and two hospital strains. Such clustering of strains from two different settings based on their genomes suggests that these strains could persist in both. WGS revealed 41 potential resistance genes on average in the hospital strains, but fewer (n=32) were detected in the community strains. In contrast, 68 virulence genes were commonly seen in strains from both sources. This study highlights the possible transmission threat to public health posed by virulent A. baumannii present in the gut of asymptomatic individuals in the community.

A review on association of fungi with the development and progression of carcinogenesis in the human body.

The role and impact of commensal and pathogenic fungi in different parts of the human body are being increasingly appreciated, unveiling the importance of such microorganisms in human health. A key function is the involvement of the mycobiota in cross-kingdom interactions within the microbiome. Any disturbance in the functionality of the microbiota could alter metabolic reactions, have a negative impact on homeostasis or induce diseases. The association of fungi with cancer development is the focus of this review. Several studies have reported direct or indirect involvement of fungal pathogens and mycobiome dysbiosis in induction of carcinogenesis. Most studies focused on cancers of the gastrointestinal tract. However, researchers are now investigating other organs, such as the skin, where the significant results obtained confirm the involvement of fungal pathogens and administration of antifungal drugs in development of cancer. This review gives an overview of the different organs affected and describes the mechanisms used by these eukaryotes or antifungals to induce oncogenesis.

Molecular typing of multi-drug resistant Candida albicans isolated from the Segamat community, Malaysia.

In the past decade, researchers have focused on the emergence of drug resistance in fungal pathogens such as Candida albicans, also considered as pathobionts that occur harmlessly in the human body but could potentially be triggered to cause diseases. The increasing rate of antifungal resistance in commensal gut fungi is alarming and should be further investigated. Here, we report seven novel MLST (Multi Locus Sequence Typing) genotypes of multi-drug resistant C. albicans isolates obtained from participants of a community study in Segamat, a district in the state of Johor, Malaysia. A total of eight C. albicans were isolated from four individuals, which were found to express high resistance against fluconazole, itraconazole, voriconazole and 5-fluorocytosine antifungals. MLST was performed to assess the clonal relatedness of these drug resistant isolates among themselves and against other strains isolated from other geographical regions. The novel MLST C. albicans sequence types suggest significant genetic changes compared to previous genotypes.

First reported case of Gilbertella persicaria in human stool: outcome of a community study from Segamat, Johor, Malaysia.

Species of fungi belonging to the order Mucorales can be found everywhere in the environment. Gilbertella persicaria, which belongs to this order, have often been isolated from fruits and in water systems. However, there has been no report of isolation of this fungus from human samples. During a gut mycobiome study, from the Segamat community, Gilbertella persicaria was isolated from a human fecal sample and was characterized through a series of morphological assessment, biochemical tests, and molecular techniques. The isolate produced a white velvety surface that turned grayish after 24 h. Although no biofilm production was observed, the results indicated that the isolate could form calcium oxalate crystals, produced urease, and was resistant to low pH. The isolate was sensitive to amphotericin but resistant to voriconazole and itraconazole. The features of this fungus that could help in its survival in the human gut are also discussed.