Selection Of Anti-fat Accumulation Probiotics On Caenorhabditis Elegans And Polyphasic Taxonomy Analysis Of Four Novel Strains

With obesity being a growing threat to human health,the role of probiotics has been paid more and more attention to,while isolating anti-fat accumulation probiotics may be able to alleviate the problem of obesity.The probiotics that have been put into use are mainly lactobacillus.Marine environment,with great microbial diversity,can be a major source for probiotics isolation.In this paper,marine bacteria isolated from the viscera of marine living oysters and Lactobacillus isolated from pickle were used to feed C.elegans to find anti-fat accumulation probiotics.In addition,there are a lot of potential novel bacteria in the ocean,four novel marine bacteria strains were identified using polyphasic methods which enriched the microbial resources.This paper has the following results:Strain ML182(Olleya marilimosa)which was able to reduce the fat accumulation of C.elegans was isolated Oysters were collected from the Weihai international bathing beach(37°31'51.09"N,122°2'43.76"E).32 bacteria strains isolated from oyster viscera,18 isolated from pickles of which 6 were lactobaccilus,8 strains from the laboratory resource collection,as well as the four novel marine strains identified were used to feed C.elegans to find anti-obesity probiotics.A marine bacteria strain ML 182(Olleya marilimosa),was able to reduce the accumulation of fat,even after autoclaving.Therefore ML 182 was selected as a candidate probiotic.Physiological and biochemical reactions of C.elegans were tested and gene of fat synthesis and degradation was analyzed after feeding marine bacteria strain ML182.Testing analysis showed that strain ML182 was non-toxic and had no effect on vital signs of C.elegans while reducing the fat accumulation.RT-qPCR results showed that the fat synthesisenzyme related genes fasn-1,pod-2,fat-5,fat-6 and fat-7 were down regulated,while acs-2,the intermediate gene of fatty acid?-oxidization was up regulated,which suggested that feeding marine bacteria ML 182 can reduce fat accumulation of C.elegans.Four novel marine bacteria strain were identified and taxonomically studied using polyphasic methods.A novel Gram-stain-negative,facultatively anaerobic bacterium,designated YH6T,was isolated from marine sediment in Weiuai,China.The sole respiratory lipoquinone was Q-8 and the major fatty acids(>10%)were C16:0 and summed feature 3(C16:1 ?7c/iso-C15:0 2-OH).The polar lipids profile of the novel strain consisted of phosphatidylglycerol,phosphatidylethanolamine,diphosphatidylglycerol,and several other unknown lipids(phospholipids,lipids,and aminophospholipids).The G+C content of the genomic DNA was 46.5 mol%.The closest type strain of strain YH6T is Vibrio variabilis(92.99%)followed by Paramoritella alkaliphila(92.55%),Pseudoalteromonas aurantia(92.20%)and Pseudoalteromonas citrea(92.20%).On the basis of the 16S rRNA gene sequence data as well as physiological and biochemical characteristics,we concluded that the starin YH6T represents a novel species of a new genus.We propose the name Motilimonas eburnea gen.nov.,sp.nov.for this new species.The type strain of the new species is YH6T(=KCTC 42594T=MCCC 1H00122T).A novel Gram-stain-negative,rod-shaped facultatively anaerobic,agarolytic bacterium,designated 017T,was isolated from Gracilaria blodgettii collected at the coast of Lingshui county,Hainan province,China.Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 017T shared the highest similarity with Teredinibacter turnerae T7902T(94.4%).The predominant polar lipids profile of the novel isolate consisted of phosphatidylglycerol,phosphatidylethanolamine,aminophospholipid and some other unknown lipids.Major cellular fatty acids(>10%)were C16:0,C18:1?7c and summed feature 3(C16:1 ?7c/iso-C15:0 2-OH),and the sole respiratory lipoquinone was Q-8.The DNA G+C content of strain 017-T was 40.2 mol%.Comparative analysis of 16S rRNA gene sequences and characterisation indicated that strain 017T represents a novel species in a new genus of the family Cellvibrionaceae,order Cellvibrionales,for which the name Agarilytica rhodophyticola gen.nov.,sp.nov.is proposed.The type strain of Agarilytica rhodophyticola is 017T(=KCTC 42584T =MCCC 1H00123 T).A novel Gram-stain-negative,rod-shaped bacterium,designated WDS2A16aT,was isolated from a solar saltern from Weihai,China.The predominant polar lipids profile of the novel isolate consisted of two unknown phospholipid phospholipids,2 unknown aminolipids and 5 unknown lipids.The DNA G+C content of strain WDS2A16aT was 47.8 mol%.Phylogenetic analysis based on 16S rRNA gene sequences(1457 nt)revealed that strain WDS2A16aT shared the highest similarity with Methylobacter marinus(90.61%).Comparative analysis of 16S rRNA gene sequences and characterisation indicated that strain WDS2A16aT represents a novel genus of the family Gammaproteobacteria,for which the name Salinibius halmophila gen.nov.,sp.nov.is proposed.The type strain of Salinibius halmophila is WDS2A16aT(=KCTC 52225T=MCCC 1H00139T).A novel Gram-stain-negative,rod-shaped,bacterium,designated E85T,was isolated from saltern of Weihai,China.Phylogenetic analysis based on 16S rRNA gene sequences(1461 nt)revealed that strain E85-T shared the highest similarity with Spiribacter salinus(97.26%)and formed a distinct lineage within the genus Spiribacter.Comparative analysis of 16S rRNA gene sequences and characterisation indicated that strain E85T represents a novel species in the genus of Spiribacter,for which the name Spiribacter halobiumsp.nov.is proposed.The type strain ofSpiribacter halobium is E85T.