Association of Obesity with Serum Leptin,Adiponectin, and Serotonin and Gut Microflora in Beagle Dogs

Background

Serotonin (5‐hydroxytryptamine, 5HT) is involved in hypothalamic regulation of energy consumption. Also, the gut microbiome can influence neuronal signaling to the brain through vagal afferent neurons. Therefore, serotonin concentrations in the central nervous system and the composition of the microbiota can be related to obesity.

Objective

To examine adipokine, and, serotonin concentrations, and the gut microbiota in lean dogs and dogs with experimentally induced obesity.

Animals

Fourteen healthy Beagle dogs were used in this study.

Methods

Seven Beagle dogs in the obese group were fed commercial food ad libitum, over a period of 6 months to increase their weight and seven Beagle dogs in lean group were fed a restricted amount of the same diet to maintain optimal body condition over a period of 6 months. Peripheral leptin, adiponectin, 5HT, and cerebrospinal fluid (CSF‐5HT) levels were measured by ELISA. Fecal samples were collected in lean and obese groups 6 months after obesity was induced. Targeted pyrosequencing of the 16S rRNA gene was performed using a Genome Sequencer FLX plus system.

Results

Leptin concentrations were higher in the obese group (1.98 ± 1.00) compared to those of the lean group (1.12 ± 0.07, P = .025). Adiponectin and 5‐hydroytryptamine of cerebrospinal fluid (CSF‐5HT) concentrations were higher in the lean group (27.1 ± 7.28) than in the obese group (14.4 ± 5.40, P = .018). Analysis of the microbiome revealed that the diversity of the microbial community was lower in the obese group. Microbes from the phylum Firmicutes (85%) were predominant group in the gut microbiota of lean dogs. However, bacteria from the phylum Proteobacteria (76%) were the predominant group in the gut microbiota of dogs in the obese group.

Conclusions and Clinical Importance

Decreased 5HT levels in obese group might increase the risk of obesity because of increased appetite. Microflora enriched with gram‐negative might be related with chronic inflammation status in obese dogs.     

Size Does Matter: Application-driven Approaches for Soil Metagenomics

Metagenomic analyses can provide extensive information on the structure, composition, and predicted gene functions of diverse environmental microbial assemblages. Each environment presents its own unique challenges to metagenomic investigation and requires a specifically designed approach to accommodate physicochemical and biotic factors unique to each environment that can pose technical hurdles and/or bias the metagenomic analyses. In particular, soils harbor an exceptional diversity of prokaryotes that are largely undescribed beyond the level of ribotype and are a potentially vast resource for natural product discovery. The successful application of a soil metagenomic approach depends on selecting the appropriate DNA extraction, purification, and if necessary, cloning methods for the intended downstream analyses. The most important technical considerations in a metagenomic study include obtaining a sufficient yield of high-purity DNA representing the targeted microorganisms within an environmental sample or enrichment and (if required) constructing a metagenomic library in a suitable vector and host. Size does matter in the context of the average insert size within a clone library or the sequence read length for a high-throughput sequencing approach. It is also imperative to select the appropriate metagenomic screening strategy to address the specific question(s) of interest, which should drive the selection of methods used in the earlier stages of a metagenomic project (e.g., DNA size, to clone or not to clone). Here, we present both the promising and problematic nature of soil metagenomics and discuss the factors that should be considered when selecting soil sampling, DNA extraction, purification, and cloning methods to implement based on the ultimate study objectives.     

不同磷肥用量对水稻土有机碳矿化和细菌群落多样性的影响

以水稻田田间定位试验为研究对象,利用三维荧光光谱技术(3D EEMs)和454测序技术,采集4个施磷水平(0、30、60、90 kg hm-2a-1)土壤,测定其有机碳矿化、溶解有机碳(DOC)组成和结构特征及细菌群落结构和丰度的变化。结果显示:水田施磷增加了土壤速效磷(Olsen-P),从而提高了土壤DOC含量,加速了有机碳的矿化速率和累积矿化量。3D EEMs结果表明,施磷分别显著增加了荧光指数和鲜度指数值1%~10%和3%~21%,而降低了腐殖化指数,且与土壤生化性质(Olsen-P、DOC和β-葡萄糖苷酶)具有显著相关性。说明施磷通过提高Olsen-P,促进了微生物源DOC的生成,同时降低了DOC的芳香化程度、分子量及腐殖化程度,从而提高了DOC生物可降解性。同时,施磷提高了细菌群落的丰度和多样性,特别是磷诱导了多种具有碳降解功能细菌的增加,从而加速了复杂有机碳的降解和甲烷氧化。此外,主成分分析表明稻田磷素施用量在30~60 kg hm-2a-1时对土壤有机碳矿化及细菌群落多样性的提高作用最为明显。因此,适度施磷能显著提高涉碳降解微生物的活性,从而提高DOC的生物可降解性,加速有机碳的矿化速率,促进稻田土壤有机碳循环。     

Impact of inorganic nitrogen additions on microbes in biological soil crusts

Many studies have shown that changes in nitrogen (N) availability affect the diversity and composition of soil microbial community in a variety of terrestrial systems, but less is known about the responses of microbes specific to biological soil crusts (BSCs) to increasing N additions. After seven years of field experiment, the bacterial diversity in lichen-dominated crusts decreased linearly with increasing inorganic N additions (ambient N deposition; low N addition, 3.5 g N m⁻² y⁻¹; medium N addition, 7.0 g N m⁻² y⁻¹; high N addition, 14.0 g N m⁻² y⁻¹), whereas the fungal diversity exhibited a distinctive pattern, with the low N-added crust containing a higher diversity than the other crusts. Pyrosequencing data revealed that the bacterial community shifted to more Cyanobacteria with modest N additions (low N and medium N) and to more Actinobacteria and Proteobacteria and much less Cyanobacteria with excess N addition (high N). Our results suggest that soil pH, together with soil organic carbon (C), structures the bacterial communities with N additions. Among the fungal communities, the relative abundance of Ascomycota increased with modest N but decreased with excess N. However, increasing N additions favored Basidiomycota, which may be ascribed to increases in substrate availability with low lignin and high cellulose contents under elevated N conditions. Bacteria/fungi ratios were higher in the N-added samples than in the control, suggesting that the bacterial biomass tends to dominate over that of fungi in lichen-dominated crusts after N additions, which is especially evident in the excess N condition. Because bacteria and fungi are important components and important decomposers in BSCs, the alterations of the bacterial and fungal communities may have implications in the formation and persistence of BSCs and the cycling and storage of C in desert ecosystems.     

Plot-scale manipulations of organic matter inputs to soils correlate with shifts in microbial community composition in a lowland tropical rain forest

Little is known about the organisms responsible for decomposition in terrestrial ecosystems, or how variations in their relative abundance may influence soil carbon (C) cycling. Here, we altered organic matter in situ by manipulating both litter and throughfall inputs to tropical rain forest soils, and then used qPCR and error-corrected bar-coded pyrosequencing to investigate how the resulting changes in soil chemical properties affected microbial community structure. The plot-scale manipulations drove significant changes in microbial community composition: Acidobacteria were present in greater relative abundance in litter removal plots than in double-litter plots, while Alphaproteobacteria were found in higher relative abundance in double-litter and throughfall reduction plots than in control or litter removal plots. In addition, the bacterial:archaeal ratio was higher in double-litter than no-litter plots. The relative abundances of Actinobacteria, Alphaproteobacteria and Gammaproteobacteria were positively correlated with microbial biomass C and nitrogen (N), and soil N and C pools, while acidobacterial relative abundance was negatively correlated with these same factors. Bacterial:archaeal ratios were positively correlated with soil moisture, total soil C and N, extractable ammonium pools, and soil C:N ratios. Additionally, bacterial:archaeal ratios were positively related to the relative abundance of Actinobacteria, Gammaproteobacteria, and Actinobacteria, and negatively correlated to the relative abundance of Nitrospira and Acidobacteria. Together, our results support the copiotrophic/oligotrophic model of soil heterotrophic microbes suggested by Fierer et al. (2007).     

Fine root decomposition,nutrient mobilization and fungal communities in a pine forest ecosystem

Despite its importance to energy flow and nutrient cycling the process of fine root decomposition has received comparatively little detailed research. Disruption of the fine root-soil interface during preparation of root litterbags for decomposition studies could affect decay rates and nutrient mobilization in part by altering the community of decay organisms. We compared rates of decomposition and nutrient release from fine roots of pine between litterbags and intact cores and characterized the fungal community in the decomposing roots. Fine root decomposition was about twice as fast overall for intact cores than litterbags, and rapid mobilization of N and P was observed for roots in cores whereas nutrients were immobilized in litterbags. Fungal communities characterized using 454 pyrosequencing were considerably different between decaying roots in intact cores and litterbags. Most interesting, taxa from ectomycorrhizal fungal orders such as Boletales, Thelephorales and Cantharellales appeared to be more common in decaying roots from cores than litterbags. Moreover, the rate of N and P mobilization from decaying fine roots was highly correlated with taxa from two orders of ectomycorrhizal fungi (Thelephorales, Cantharellales). Although we caution that DNA identified from the decaying roots cannot be conclusively ascribed to active fungi, the results provide tentative support for a significant role of ectomycorrhizal fungi in decomposition and nutrient mobilization from fine roots of pine.     

Members of soil bacterial communities sensitive to tillage and crop rotation

Pyrosequencing was used to study the effect of rotation and tillage on total bacterial communities. We designed primers to the bacterial 16s rDNA and amplified DNA from soil samples from a long-term tillage/rotation trial in Kansas for two seasons. The 2 × 2 factorial trial had two rotation treatments (wheat-wheat and wheat-soybean) and two tillage treatments (conventional and no-till). A total of 20,180 16s rDNA sequences were generated and 2337 operational taxonomic units (OTUs) were assembled using a 97% similarity cut-off. The phylum Proteobacteria represented 38% of 299 identified taxa. The second most abundant phylum was Acidobacteria, making up 20% of the sequences, the majority of which were Acidobacteria Group 1. The phyla Actinobacteria and Gemmatimonadetes comprised 12% and 3.5% of the sequences. Other groups detected included TM7, Nitrospira, Verrucomicrobia, and Bacteroidetes. Some clusters of Acidobacteria Group 1 were more frequent in continuous wheat versus wheat-soybean rotation, some Acidobacteria Group 2 were more frequent in no-till, and some Acidobacteria Group 4 were more frequent in wheat-soybean rotation. These results were validated by quantitative real-time PCR. Pyrosequencing provided taxonomic information about the overall bacterial community, and detected community shifts resulting from different cropping practices.     

用焦磷酸测序技术检测凡纳滨对虾组织蛋白酶基因单核苷酸多态性

焦磷酸测序是一种新型的DNA测序技术,近年来开始应用于单核苷酸多态性(SNPs)检测.组织蛋白酶基因(Cathepsin-L,CTSL)在对虾的蜕皮周期中起重要的作用.本研究利用焦磷酸测序技术,对96个凡纳滨对虾的CTSL基因序列(GnBank登录号:AY366355)的C681G SNP位点进行检测分型.结果发现C/C、C/G和G/G基因型,频率分别是0.81、0.16和0.03,频率分布符合Hardy-Weinberg平衡.统计分析结果表明在该群体中C681G SNP和体重关系不显著(P>0.05).研究结果显示焦磷酸测序技术是一种高效的SNP检测技术,可以在对虾的遗传研究中发挥重要作用.     

马氏珠母贝转录组数据的重新组装

对本实验室和其他学者已经发表的马氏珠母贝外套膜和珍珠囊的454焦磷酸测序转录组数据以及NCBI中的EST数据进行了重组装、同源序列比对和GO注释分析。重组装获得了30266个contig,平均长度522 bp,其中最长的contig为4144 bp;NR数据库BLAST分析获取了2310个相似性contig(E≤10?5),其中1902个在模式动物(人、小鼠、海胆、线虫、斑马鱼、果蝇或者太平洋牡蛎和珠母贝)中可以查找到同源序列。基因注释(GO)结果表明,注释到生物学过程的3846个contig可分为23个亚类,其中代谢过程蛋白、细胞过程蛋白和生物学调控蛋白为contig数量最多的3个亚类;注释到分子功能的4601个contig共分为11个亚类,其中结合蛋白的contig数量最多,其次是具有催化活性的蛋白和结构分子活性蛋白;注释到细胞成分的2992个contig分为17个亚类,其中细胞和细胞组成部分的contig数量最多,其次分别是细胞器和大分子复合体contig数量。本研究结果有助于开展外套膜和珍珠囊特异性表达基因的大规模筛选。