24 25 26 Email addresses: 27 HFA fauzanahmad@ump.edu.my 28 JLC jcame@food.ku.dk 29 ŁK krych@food.ku.dk 30 BK bzo@food.ku.dk 31 WK wk@envs.au.dk 32 RLB r.bechshoeft@gmail.com 33 SR s.reitelseder@gmail.com 34 GWH grith.westergaard.hoejfeldt.01@regionh.dk 35 SBE se@food.ku.dk 36 LH l.holm@bham.ac.uk 37 KF karoline.faust@kuleuven.be 38 DSN dn@food.ku.dk 39 40 41 42 43 44 45 ABSTRACT 46 Gut microbial dysbiosis have been in the etiology of a number of diseases, yet 47 the presence of fungal communities and their possible association with host health are 48 little understood. This study attempts to identify gut microbial fungal associations 49 with the progression of atherogenic dyslipidemia in a population of older adults by 50 investigating the interplay between dietary intake, gut mycobiome composition, 51 plasma and fecal metabolome and anthropometric/body-composition measurements of 52 100 Danes aged 65 to 81 (69.57 3.64) years. The gut mycobiome composition were 53 determined by high-throughput sequencing of internal transcribed spacer (ITS2) gene 54 amplicons, while the plasma and fecal metabolome was determined by GC-TOF-MS. 55 The gut microbiome of the subjects investigated is home to three main eukaryotic 56 phyla, namely Ascomycota, Basidiomycota and Zygomycota, with genera 57 Penicillium, Candida, and Aspergillus being particularly common. 58 Hypertriglyceridemia was associated with fewer observed fungal species, and Bray-59 Curtis dissimilarity matrix-based analysis showed significant (P<0.05) clustering 60 according to fasting levels of circulating plasma triglycerides (Tg) and very low-61 density lipoprotein (VLDL) cholesterol fasting levels, respectively. Interestingly, 62 neither hypertriglyceridemia nor elevated VLDL levels were reflected in the 63 prokaryotic component of the gut microbiome as determined by 16S rRNA gene 64 amplicon sequencing. Higher levels of Tg and VLDL cholesterol significantly 65 associates with increased relative abundance of genus Penicillium, possibly mediated 66 by a higher dietary fat intake (ANOVA, P<0.05), and Aspergillus and Guehomyces 67 were positively associated with SCFAs groups. Collectively, these findings suggest 68 that in older adults' gut mycobiome dysbiosis is associated with hypertriglyceridemia, 69 a known risk factor for development of cardiovascular disease. 70 71 Keywords: older-adults, hypertriglyceridemia, dysbiosis, gut mycobiome, host 72 metabolome, triglyceride, VLDL and dietary fat intake 73 74 75 93 dyslipidaemia in both men and women older than 65 years, as compared to younger 94 individuals [16]. Further, elevated triglycerides (Tg) and very low density level 95 (VLDL) cholesterol levels have been associated with subclinical atherosclerosis and 96 dubbed as independent risk factors for CVD [17]. Several large studies suggest that 97 hypertriglyceridemia is related to increased levels of remnant lipoproteins in 98 promoting atherogenesis [18], [19]. The possible mechanisms for this association 99 include excessive free fatty acid release, product...