Gut Mycobiome Dysbiosis is Linked to Hypertriglyceridemia Among Home Dwelling Elderly Danes

Abstract

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...

The role of gut mycobiome in health and diseases

“…Fungal community increased with age in infancy but decreased when they grow up to young adults ( Figure 2 ). 88 However, gut mycobiota alteration in elder subjects was less reported, 114 and most of the older adults studied were reported with known diseases including hypertriglyceridemia (HG), 115 Alzheimer’s disease (AD), 51 , 116 and type 2 diabetes mellitus (T2DM). 19 In Denmark, gut mycobiome analysis was performed in 100 elderly participants (70 with normotriglyceridemia versus 30 with hypertriglyceridemia) aged 65–81 years by ITS 2 amplicon sequencing on their fecal samples.…”

“…Penicillium, Candida , and Aspergillus were the top three genera among the elderly Danes, and genus Penicillium was strongly correlated with the hypertriglyceridemia. 115 Another study on 17 older subjects (11 mild cognitive impairment versus 6 cognitively normal) with average age of 64.6 years showed that Saccharomyces, Candida , and Aspergillus were major fungal genera in their fecal samples through ITS 1 sequencing. Higher abundance of Botrytis, Kazachstania, Phaeoacremonium , and Cladosporium and decreased abundance of Meyerozyma at genus level were found in the gut of patients with mild cognitive impairment compared with controls.…”

Metadata Analysis For Gut Microbiota between Indoor and Street Cats of Malaysia

“…Amplicon libraries were created using two-steps custom PCR protocols targeting the V4 region of 16s rRNA as described previously: 515fB (5′ GTGYCAGCMGCCGCGGTAA 3′) and 806rB (5′ GGACTACNVGGGTWTCTAAT 3′) [11]. Briefly, the 16S rRNA V4 region was amplified using OneTaq® 2X Master Mix (NEB, Ipswich, USA) from the extracted gDNA using the primer pair 515F-806R [1] containing a partial Illumina Nextera adapter in their 5' end. The PCR condition used was 94 o C -30 seconds followed by 30 cycles of 94 o C -15 sec, 50 o C -15 sec and 68 o C -30 seconds.…”

“…Microbiota is known as a microbial community that shares either commensalism, mutualistic or pathogenic relationships. Over the years, studies on the composition of gut bacteria have focused on human models [1], [2] with lesser attention on other kingdoms like animal, plant, and the environment. Recently, studies on mammals such as dogs and cats are getting highlights as these animals are kept as human beings' pets and inhibit the same living quarters.…”

Full-length ITS1 Genes Amplicon Analysis of Gut Mycobiome Using Nanopore Flongle Sequencing Reveals Unique Fungal Communities between Indoor and Outdoor Felines in Malaysia

“…The gastrointestinal tract (GIT) is an environment that hosts complex microbial ecosystems dominated by bacteria and other organisms such as fungus, Archaea, viruses and protozoa, collectively known as gut microbiomes [1]. The fungal community found in the GI tract is known as gut mycobiota, and like their bacteria counterpart, they have a profound impact on health and disease [2] by playing an important role in maintaining gut functioning and immune development [3], [4]. Gut microbiota is involved in energy harvesting, storage, and regulation of host metabolic functioning and is essential for the production and absorption of metabolites like short-chain fatty acids [5] and other metabolites necessary for health and survival [6].…”