16S-rDNA

  • 文章类型: Journal Article
    绿僵菌属。已经成为化学农药的替代品,用于保护作物免受虫害。这里,我们调查了斜纹夜蛾感染黄花绿僵菌后三个不同时间点的中肠微生物群落和代谢产物。Litura的先天免疫系统被多酚氧化酶水平激活,羧酸酯酶,多功能氧化酶,谷胱甘肽S-转移酶活性显著增加。暴露于真菌病原体也改变了宿主中肠的细菌丰度和多样性,这些变化取决于曝光后经过的时间。在所有测试的时间点,与对照样品相比,我们在处理的样品中鉴定了更多的操作分类单位。总共鉴定出372种代谢物,在治疗后的3个时间点,在治疗组和对照组之间鉴定出88、149和142种差异积累的代谢物(DAMs),分别。基于不同时间点DAMs对Flavoviride感染的反应变化,并显着丰富了KEGG途径,我们推测,“酪氨酸代谢,“\”半乳糖代谢,\“\”ATP结合盒转运蛋白,神经活性配体-受体相互作用,嘌呤代谢,精氨酸和脯氨酸代谢,β-丙氨酸代谢,\"\"溶酶体,“碳代谢”和“碳代谢”可能参与代谢水平的防御反应。对16S-rDNA和代谢组学数据的综合途径水平分析说明了斜纹链球菌和中肠微生物对黄花支原体感染的代谢反应之间的联系和相互依赖性。这项工作强调了昆虫-病原体相互作用综合分析的价值,为这些相互作用的关键微生物和代谢决定因素的未来研究提供了框架,为黄维脲的可持续利用奠定了理论基础。
    Metarhizium spp. have emerged as an alternative to chemical pesticides for protecting crops from insect pest. Here, we investigated midgut microbial community and metabolites of Spodoptera litura at three different timepoints after infection with Metarhizium flavoviride. The innate immune system of S. litura was activated with levels of polyphenol oxidase, carboxylesterase, multifunctional oxidase, and glutathione S-transferase activity significantly increasing. Exposure to the fungal pathogen also altered bacterial abundance and diversity in host\'s midgut, and these changes varied depending on the time elapsed since exposure. We identified more operational taxonomic units in the treated samples as compared to the control samples at all tested time points. A total of 372 metabolites were identified, and 88, 149, and 142 differentially accumulated metabolites (DAMs) were identified between the treatment and control groups at 3 timepoints after treatment, respectively. Based on the changes of DAMs in response to M. flavoviride infection at different timepoints and significantly enriched KEGG pathways, we speculated that \"tyrosine metabolism,\" \"galactose metabolism,\" \"ATP-binding cassette transporters,\" \"neuroactive ligand-receptor interaction,\" \"purine metabolism,\" \"arginine and proline metabolism,\" \"beta-alanine metabolism,\" \"lysosome,\" and \"carbon metabolism\" may participate in the metabolic-level defense response. An integrated pathway-level analysis of the 16S-rDNA and metabolomic data illustrated the connections and interdependencies between the metabolic responses of S. litura and the midgut microorganisms to M. flavoviride infection. This work emphasizes the value of integrated analyses of insect-pathogen interactions, provides a framework for future studies of critical microorganisms and metabolic determinants of these interactions, establishes a theoretical basis for the sustainable use of M. flavoviride.
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  • 文章类型: Journal Article
    蜱是与多种脊椎动物宿主相关的嗜血体外寄生虫。在这个群体中,Ixodidae家族脱颖而出,其中Ixodes属在世界范围内至少包含245种,其中55种存在于新热带地区。Ixodesaffinis,1899年描述的一种蜱,从美国南锥体到美国都有广泛的分布。然而,因为它的描述,据报道,其种群之间存在形态变异性。此外,已经尝试使用线粒体标记来阐明其作为物种复合体的状态,但主要在南美洲和中美洲的限制人口中。因此,缺乏与新热带和近地带之间过渡区域种群有关的信息。由于这些原因,这项研究的目的是评估整个美洲I.affinis的遗传多样性和结构,并汇编所有已发表的I.affinis在美国的记录,阐明宿主与寄生虫的关系并确定其地理分布。为此,一个系统发育,和AMOVA分析进行了评估遗传结构的样品通过实地工作在南卡罗来纳州,美国和尤卡坦,墨西哥。从16S区的片段中检索到总共86个序列。系统发育和遗传结构分析表明,在地理和遗传上具有较高的分支支持和Fst值,所有这些都具有统计学意义。获得的结果支持以下假设:I.affinis它对应于四个物种的复合物,这必须通过未来的形态学比较来验证。
    Ticks are hematophagous ectoparasites associated with a wide range of vertebrate hosts. Within this group, the Ixodidae family stands out, in which the Ixodes genus contains at least 245 species worldwide, from which 55 species are present in the Neotropical region. Ixodes affinis, a tick described in 1899, has a wide distribution from the Southern Cone of America to the United States. However, since its description, morphological variability has been reported among its populations. Furthermore, attempts have been made to clarify its status as a species complex using mitochondrial markers, but mainly in restricted populations of South and Central America. Thus, information related to populations of the transition region between the Neotropical and Nearctic zones is lacking. For these reasons, the objectives of the study were to evaluate the genetic diversity and structure of I. affinis across the Americas and to compile all the published records of I. affinis in America, to elucidate the host-parasite relationships and to identify their geographical distribution. For this, a phylogeny, and AMOVA analyses were performed to assess the genetic structure of samples obtained by field work in South Carolina, USA and Yucatán, Mexico. A total of 86 sequences were retrieved from a fragment of the 16S region. Phylogeny and genetic structure analysis showed four groups that were geographically and genetically related with high branch support and Fst values, all of them statistically significant. The results obtained support the hypothesis that I. affinis it corresponds to a complex of four species, which must be validated through future morphological comparisons.
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  • 文章类型: Journal Article
    使用内生细菌控制番茄的早期疫病是一种生态友好且可持续的方法,可以管理由格格孢菌引起的这种常见真菌病,交替菌,还有弯孢菌.内生细菌是生活在植物组织内而不造成伤害的微生物,可以帮助保护宿主植物免受病原体的侵害。在这项工作中,从番茄健康植物中分离出的20株内生细菌对引起早期疫病的病原真菌进行了体外检测。在20个测试的分离株中,三个(B4,B7和B17)被认为是对抗真菌病原体生长的有效分离株。这三个分离株被识别为阴沟肠杆菌HS-6(B4),gessardii假单胞菌HS-5(B7),和地中海假单胞菌HS-4(B17)使用16s-rDNA测序。在体外测试了不同浓度的20、40和60%的细菌培养物扩张剂对病原真菌发育的拮抗作用。在所有细菌培养滤液中,致病性分离株的最低干重为60%。在所有培养滤液中,酚类化合物的峰面积最大。在温室条件下,与其他处理相比,阴沟肠球菌及其培养滤液的番茄早疫病的病情严重程度最低。实时PCR用于检查防御反应基因β-1.3葡聚糖酶基因在感染病原真菌(对照)的番茄植株中的表达模式及其与有效生物防治剂的关系(E。泄殖腔)。在接种C.lunata和E.cloacae感染的植物三天后,该基因的表达显着增加,而在接种A.alternata后五天达到最大值。A.solani和阴沟肠杆菌。我们的研究得出结论,内生细菌分离阴沟菌可以被认为是预防番茄早疫病的有前途的生物防治剂。
    Controlling early blight of tomatoes using endophytic bacteria is an eco-friendly and sustainable approach to manage this common fungal disease caused by Alternaria solani, Alternaria alternata, and Curvularia lunata. Endophytic bacteria are microorganisms that live inside plant tissues without causing harm and can help protect the host plant from pathogens. In this work, twenty endophytic bacterial isolates from tomato healthy plants were tested against pathogenic fungal isolates that caused early blight disease in vitro. Out of the 20 tested isolates, three (B4, B7, and B17) were considered effective isolates against the growth of fungal pathogens. The three isolates were recognized as Enterobacter cloacae HS-6 (B4), Pseudomonas gessardii HS-5 (B 7), and Pseudomonas mediterranea HS-4 (B17) using 16s-rDNA sequencing. Different concentrations of bacterial cultural diltrates at 20, 40, and 60% were tested for their antagonistic effects on the development of pathogenic fungi in vitro. The lowest dry weights of pathogenic isolates in all bacterial culture filtrates were discovered at 60%. In all culture filtrates, phenolic compounds showed the largest peak area. Under greenhouse conditions, the least disease severity of tomato early blight was found for E. cloacae and its culture filtrate compared to other treatments. Real-time PCR was used to examine the expression pattern of the defense response gene β-1.3 glucanase gene in infected tomato plants with pathogenic fungi (control) as well as its relations with efficient biocontrol agent (E. cloacae). The expression of the gene increased substantially and significantly after three days from the inoculation-infected plants with C. lunata and E. cloacae while it reached the maximum after five days from the inoculation with A. alternata, A. solani and E. cloacae. Our study concluded that the endophytic bacterial isolate E. cloacae can be considered a promising biocontrol agent for preventing tomato early blight.
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  • 文章类型: Journal Article
    患有糖尿病前期的男性通常表现出伴随的低质量精子产生甚至不育,迫切需要改进治疗方案的问题。在这项研究中,我们建立了一个与生精缺陷相关的饮食诱导的前期糖尿病绵羊模型,并探索了可能的潜在代谢原因.
    我们比较了饲喂正常饮食的雄性绵羊与那些由丰富饮食诱发前驱糖尿病的绵羊,以及第三组由褪黑激素补充丰富饮食的绵羊。只有丰富的饮食组有糖尿病前期的症状,在这些羊身上,我们发现精子发生受损,其特征是圆形精子细胞发育受阻,睾丸凋亡细胞数量增加。比较三组的肠道微生物组和肠道消化代谢组,发现糖尿病前期绵羊的微生物群的分类组成存在显着差异,和改变的代谢组,其最重要的特征是鞘氨醇代谢改变;在血液和睾丸中也发现了鞘氨醇升高。褪黑素的给药减轻了糖尿病前期的症状,包括精子发生受损的精子发生,同时恢复更正常的微生物群和鞘氨醇的代谢水平。糖尿病前期绵羊的粪便微生物移植诱导受体小鼠鞘氨醇水平升高和精子发生受损,表明肠道微生物群在这些表型中的因果作用。
    我们的研究结果指出鞘氨醇在糖尿病前期绵羊精子发生破坏中的关键作用,并表明它可能是一个有用的疾病标志物;此外,褪黑激素是治疗前驱糖尿病引起的男性不育的潜在益生元。
    Men with prediabetes often exhibit concomitant low-quality sperm production or even infertility, problems which urgently require improved therapeutic options. In this study, we have established a sheep model of diet-induced prediabetes that is associated with spermatogenic defects and have explored the possible underlying metabolic causes.
    We compared male sheep fed a normal diet with those in which prediabetes was induced by a rich diet and with a third group in which the rich diet was supplemented by melatonin. Only the rich diet group had symptoms of prediabetes, and in these sheep, we found impaired spermatogenesis characterized by a block in the development of round spermatids and an increased quantity of testicular apoptotic cells. Comparing the gut microbiomes and intestinal digest metabolomes of the three groups revealed a distinctive difference in the taxonomic composition of the microbiota in prediabetic sheep, and an altered metabolome, whose most significant feature was altered sphingosine metabolism; elevated sphingosine was also found in blood and testes. Administration of melatonin alleviated the symptoms of prediabetes, including those of impaired spermatogenesis, while restoring a more normal microbiota and metabolic levels of sphingosine. Fecal microbiota transplantation from prediabetic sheep induced elevated sphingosine levels and impaired spermatogenesis in recipient mice, indicating a causal role of gut microbiota in these phenotypes.
    Our results point to a key role of sphingosine in the disruption of spermatogenesis in prediabetic sheep and suggest it could be a useful disease marker; furthermore, melatonin represents a potential prebiotic agent for the treatment of male infertility caused by prediabetes.
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  • 文章类型: Journal Article
    背景:尽管有很多假设,克罗恩病(CD)的发病机制至今仍不完全清楚。独家肠内营养(EEN)是治疗活动性CD的常规措施。我们旨在研究EEN对来自微生物代谢组学的活性CD患者的影响。
    方法:采用16S-rDNA测序技术和气相色谱-质谱分析研究EEN过程中肠道菌群和粪便短链脂肪酸(SCFA)的变化。
    结果:7名CD患者,谁主持了EEN,在本研究中成功随访。为期8周的EEN缓解了患有活动性CD的受试者的病情,如红细胞沉降率(ESR)显着降低(P=0.018)所示,C反应蛋白(CRP)(P=0.028),克罗恩病活动指数(CDAI)(P=0.018)。在EEN治疗8周后,受试者的营养得到改善,这与身体脏乱指数(BMI)(P=0.018)和血清白蛋白(ALB)(P=0.018)水平的增加有关。此外,我们的调查显示,Firmicutes的丰度显着增加,同时变形杆菌的水平降低。关于属,五种细菌包括Ruminococus(P=0.01),落叶松科(P=0.02),anaerotruncus(P=0.04),Flavonifractor(P=0.04),和新鞘氨醇(P=0.05)的丰度显着增加。这伴随着粪便短链脂肪酸水平的相对变化。此外,我们通过结合这5个显著不同的属,成功构建了一个稳定的模型来预测EEN对CD患者的治疗效果(AUC=0.9598).
    结论:结果表明,EEN可以通过调节肠道菌群和影响粪便短链脂肪酸的表达水平,减轻活动性CD患者的病情和营养。
    BACKGROUND: Although there are many hypotheses, the pathogenesis of Crohn\'s disease (CD) is not completely clear so far. Exclusive enteral nutrition (EEN) is a routine measure in the treatment of active CD. We aimed at investigating the impact of EEN on patients with active CD from microbial metabolomics.
    METHODS: 16S-rDNA sequencing technology and gas chromatography-mass spectrometer analysis were employed to investigate the modification of the intestinal flora and fecal short-chain fatty acid (SCFA) during the EEN.
    RESULTS: Seven patients with CD, who conducted EEN, were followed up successfully in the present study. The 8-week EEN resulted in a remission of the condition of subjects with active CD, as revealed by a significant decrease in erythrocyte sedimentation rate (ESR) (P = 0.018), C-reactive protein (CRP) (P = 0.028), and Crohn\'s disease activity index (CDAI) (P = 0.018). The nutrition of the subjects was improved after an 8-week treatment course with EEN, which was associated with an increase in body mess index (BMI) (P = 0.018) and serum albumin (ALB) (P = 0.018) levels. Furthermore, our investigations revealed a significantly increased abundance of Firmicutes paralleled by decreased levels of Proteobacteria. With respect to the genus, five species of bacteria including Ruminococcus (P = 0.01), Lachnospiraceae (P = 0.02), Anaerotruncus (P = 0.04), Flavonifractor (P = 0.04), and Novosphingobium (P = 0.05) showed significantly increased abundance. This was accompanied by relative changes in fecal short-chain fatty acids levels. Moreover, we successfully constructed a stable model by combining these five significantly different genera to predict the therapeutic effect of EEN on patients with CD (AUC = 0.9598).
    CONCLUSIONS: The findings indicated that EEN can alleviate the condition and the nutrition of patients with active CD by regulating the intestinal flora and influencing the expression level of fecal short-chain fatty acids.
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  • 文章类型: Journal Article
    OBJECTIVE: This study aimed to explore the effects of Psidium guajava L. extracts on mice diarrhea model and changes in their intestinal microflora diversity.
    METHODS: Mice diarrhea model was constructed with Folium Sennae. and The therapeutic effects of Psidium guajava L. extracts on diarrhea were evaluated by loose stools rates, diarrhea rates, diarrhea index and motor functions of intestine.
    RESULTS: Our results showed that all three kinds of extracts from Psidium guajava L. had inhibitory effects on Folium Sennae -induced diarrhea model. The proportion of Bacteroidetes is lower in Group DF (diarrhea model group) compared with Group B (blank group), while abundance of Deferribacteraceae was found in Group DF. The proportion of Deferribacteraceae lowered in Group DTF (diarrhea treatment group) treated with extracts from Psidium guajava L.
    CONCLUSIONS: However, mechanisms underlying the therapeutic effect of Psidium guajava L. and the changes in intestinal microflora still await further exploration.
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