Effects of probiotics supplementation on physiology of silkworm larvae and nutrient characteristics of silkworm pupa

<p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d11508354e174"> <b>[Objective]</b> This study aimed to investigate the effects of probiotic supplementation on the physiology of silkworm larvae and the nutrient characteristics of silkworm pupae, which could provide theoretical basis for the application of probiotics in silkworm rearing and the development of high-quality silkworm pupae food products. </p><p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d11508354e179"> <b>[Method]</b> Fifthinstar silkworm larvae were fed mulberry leaves sprayed with <i>Lactobacillus rhamnosus</i> GIM 1.325 (LR group) or <i>Lactobacillus casei</i> GIM 1.411 (LC group), while the control group (CON) was fed with mulberry leaves sprayed with an equal amount of MRS broth medium. Hemolymph, midgut supernatant, fat body and whole intestine (including contents) were sampled. Mortality rates of larvae and pupae were recorded on day 6, and the survival rate of fifth-instar larvae and cocoon layer rate were calculated. The reagent kit was used to measure intestinal α-amylase, lipase, and trypsin activities, as well as hemolymph total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD) activity and 1, 1-dipheny-l-2-picrylhydrazyl (DPPH) radical scavenging ability. Real-time fluorescence quantitative PCR was used to analyze the relative expression levels of antimicrobial peptide genes in the fat body. Intestinal flora was analyzed via operational taxonomic unit (OTU) clustering and alpha diversity indexes using the Illumina HiSeq™ 2500 platform, and pupal nutrient composition was assessed. </p><p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d11508354e190"> <b>[Results]</b> The survival rate of fifth-instar larvae silk worm in the LR and LC groups was significantly higher than that in the CON group ( <i>P</i><0.05, the same below). From day 1 to day 6 of probiotic supplementation in fifth-instar larvae silk worm, α-amylase activity of silkworm intestine in the LR and LC groups was significantly lower than in the CON group, while trypsin activity was significantly higher than in the CON group. Except on day 5, lipase activity in the LR and LC groups was significantly higher than in the CON group. Except for day 2 and day 6 the total antioxidant capacity of silkworm hemolymph in the LR group was significantly higher than in the CON group. Except for day 1 and day 5, the total superoxide dismutase activity in the LR group was significantly higher than that in the CON group. Except for the day 3 and day 6, the total antioxidant energy in the LC group was significantly lower than that in the CON group. From the day 1 to day 3, the total superoxide dismutase activity in the LC group was significantly higher than that in the CON group. The free radical scavenging ability of DPPH in both the LR and LC groups was significantly higher than that in the CON group from the day 2 to day 6. The results of real-time fluorescence quantitative PCR showed that compared with the CON group, the relative expression levels of antimicrobial peptide genes in silkworm fat body tissue <i>BmAttacin1</i>, <i>BmCecropinB6</i>, <i>BmDefensinA</i> and <i>BmLebocin3</i> in the LR and LC groups were significantly increased on day 4 and day 6 after feeding, the relative expression level of <i>BmGloverin2</i> gene in the LC group was significantly increased on the day 2, day 4 and day 6 after feeding, and the relative expression level of <i>BmMoricin</i> gene in the LR and LC groups was significantly increased on day 2, day 4 and day 6 after feeding. OTU clustering analysis indicated that, except day 4 after supplementation, the number of OTU was significantly lower in the probiotic-fed groups than in the control group at the same sampling time. MetaCyc pathway functional prediction results indicated that the relative abundance of functional genes involved in carbohydrate biosynthesis was significantly lower in the probiotic-fed groups than in the control group at the same sampling time. Nutrient analysis of pupae showed that crude protein content was significantly increased, while total sugar content was significantly decreased in the LR and LC groups compared with the CON group. </p><p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d11508354e217"> <b>[Conclusion]</b> Probiotic supplementation improves the survival rate and nutrient digestion efficiency of fifthinstar silkworm larvae. It can enhance the immunity of silkworm by regulating the total antioxidant capacity of hemolymph, the activity of superoxide dismutase, the DPPH free radical scavenging ability and the relative expression levels of antimicrobial peptide genes, and also improve the nutritional composition of silkworm pupae. </p><p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="first" dir="auto" id="d11508354e223"> <b>摘要: 【目的】</b> 探究益生菌添食对家蚕幼虫生理及蚕蛹养分特征的影响, 为益生菌在家蚕饲养中的应用及高品质 蚕蛹食品的开发提供理论参考。 <b>【方法】</b>以家蚕5龄幼虫为研究对象, 添食喷洒鼠李糖乳杆菌 ( <i>Lactobacillus rhamnosus</i> GIM 1.325) 和干酪乳杆菌 ( <i>Lactobacillus casei</i> GIM 1.411) 的桑叶, 分别设为LR组和LC组, 对照组 (CON) 添食喷洒等 量稀释的MRS肉汤培养基的桑叶。取血淋巴、中肠上清液、脂肪体和全肠 (含内容物) 。记录添食后第6 d家蚕幼虫和 蚕蛹死亡数, 计算家蚕5龄幼虫存活率和茧层率。使用试剂盒检测肠道α-淀粉酶、脂肪酶和胰蛋白酶活力与血淋巴总 抗氧化能力、总超氧化物歧化酶活力和1,1-二苯基-2-三硝基苯肼 (DPPH) 自由基清除能力。采用实时荧光定量 PCR检测脂肪体组织抗菌肽基因相对表达量。利用Illumina HiSeq™ 2500平台对肠道菌群进行操作分类单元 (OTU) 聚类分析和Alpha多样性指数分析, 并检测蚕蛹营养成分。 <b>【结果】</b>LR和LC组家蚕5龄幼虫存活率显著高于 CON组 ( <i>P</i><0.05, 下同) 。家蚕5龄幼虫添食益生菌后第1~6 d, LR和LC组家蚕肠道α-淀粉酶活力均显著低于CON组, 胰蛋白酶活力均显著高于CON组, 除第5 d外, LR和LC组脂肪酶活力均显著高于CON组;除第2和第6 d外, LR组家蚕 血淋巴总抗氧化能力均显著高于CON组, 除第1和第5 d外, LR组总超氧化物歧化酶活力均显著高于CON组, 除第3和 第6 d外, LC组总抗氧化能均显著低于CON组, 在第1~3 d, LC组总超氧化物歧化酶活力均显著高于CON组, LR和 LC组DPPH自由基清除能力在第2~6 d均显著高于CON组。实时荧光定量PCR结果显示, 与CON组相比, LR和LC组 家蚕脂肪体组织抗菌肽基因 <i>BmAttacin1</i>、 <i>BmCecropinB6</i>、 <i>BmDefensinA</i>和 <i>BmLebocin3</i>相对表达量在添食后第4和第6 d 显著升高, LC组 <i>BmGloverin2</i>基因相对表达量在添食后第2、第4和第6 d均显著升高, LR和LC组 <i>BmMoricin</i>基因相对 表达量在添食后第2、第4和第6 d均显著升高。OTU聚类分析结果显示, 除添食后第4 d外, 同一取样时间下益生菌添 食组OTU数量均显著低于对照组。MetaCyc通路功能预测分析结果显示, 同一取样时间下, 参与碳水化合物生物合成 的功能基因相对丰度显著低于对照组。蚕蛹营养成分分析结果显示, 与CON组相比, LR和LC组蚕蛹粗蛋白含量显著 升高, 总糖含量显著降低。 <b>【结论】</b>益生菌添食能提高家蚕5龄幼虫存活率和对营养物质的消化能力, 并通过调节血淋 巴总抗氧化能力、超氧化物歧化酶活力、DPPH自由基清除能力和抗菌肽基因的相对表达量来增强家蚕免疫力, 还能 改善蚕蛹营养成分。 </p>

Usefulness of microsatellite loci for differentiating between <i>Dibothriocephalus dendriticus</i> and <i>Dibothriocephalus ditremus</i> (Cestoda: Diphyllobothriidea) <span class="so-article-trans-title" dir="auto"> Translated title: Utilité des loci microsatellites pour différencier <i>Dibothriocephalus dendriticus</i> et <i>Dibothriocephalus ditremus</i> (Cestoda : Diphyllobothriidea) </span>

<p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="first" dir="auto" id="d1713179e211">Differentiating between two diphyllobothriid tapeworms <i>Dibothriocephalus dendriticus</i> and <i>Dibothriocephalus ditremus</i> is complicated due to their morphological plasticity, intraspecific variability and a wide range of common hosts. The aim of this study was to develop a species-specific PCR-based method for single-step discrimination between <i>D. dendriticus</i> and <i>D. ditremus</i>. Intraspecific variation and interspecific differences were analysed in subunits/spacers of nuclear rRNA genes and protein-coding genes of mitochondrial DNA. In addition, the specificity of primers designed for the amplification of microsatellite loci in <i>D. dendriticus</i> was tested on <i>D. ditremus</i> DNA. Due to high identity within the rRNA gene in these species, no suitable DNA regions could be identified for the design of the species-specific primers. A higher level of interspecific differences was detected in the mitochondrial <i>cox</i>1 and <i>cob</i> genes, in which regions containing species-specific mutations were chosen for the design of <i>D. dendriticus</i>- and <i>D. ditremus</i>-specific primers. However, their specificity was not confirmed, as the <i>D. dendriticus</i>-specific primers also annealed to <i>D. ditremus</i> DNA and <i>vice versa</i>. Of the 15 primer pairs designed for the amplification of microsatellite loci in <i>D. dendriticus,</i> 13 primer pairs also annealed to <i>D. ditremus</i> DNA. Only two primer pairs, which amplify the Dd_8 and Dd_33 loci have been proven to be <i>D. dendriticus</i>-specific. The effectiveness and high reproducibility of the Dd_8 primers were validated on ~3,500 <i>D. dendriticus</i> and <i>D. ditremus</i> plerocercoids from Iceland and Norway. These primers are recommended for future molecular differentiation between both <i>Dibothriocephalus</i> species. </p><p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="first" dir="auto" id="d1713179e274">La différenciation entre deux cestodes Diphyllobothriidae, <i>Dibothriocephalus dendriticus</i> et <i>Dibothriocephalus ditremus</i>, est complexe en raison de leur plasticité morphologique, de leur variabilité intraspécifique et de la grande diversité de leurs hôtes communs. L’objectif de cette étude était de développer une méthode de PCR spécifique à chaque espèce pour la discrimination en une seule étape entre <i>D. dendriticus</i> et <i>D. ditremus</i>. La variation intraspécifique et les différences interspécifiques ont été analysées dans les sous-unités/espaceurs des gènes de l’ARNr nucléaire et dans les gènes codant pour des protéines de l’ADN mitochondrial. De plus, la spécificité des amorces conçues pour l’amplification des loci microsatellites chez <i>D. dendriticus</i> a été testée sur l’ADN de <i>D. ditremus</i>. En raison de la forte identité au sein du gène d’ARNr chez ces espèces, aucune région d’ADN appropriée n’a pu être identifiée pour la conception d’amorces spécifiques à l’espèce. Un niveau plus élevé de différences interspécifiques a été détecté dans les gènes mitochondriaux <i>cox1</i> et <i>cob</i>, dans lesquels des régions contenant des mutations spécifiques à l’espèce ont été choisies pour la conception d’amorces spécifiques à <i>D. dendriticus</i> et <i>D. ditremus</i>. Cependant, leur spécificité n’a pas été confirmée, car les amorces spécifiques à <i>D. dendriticus</i> s’hybrident également à l’ADN de <i>D. ditremus</i> et <i>vice versa</i>. Sur les 15 paires d’amorces conçues pour l’amplification des loci microsatellites chez <i>D. dendriticus</i>, 13 paires d’amorces s’hybrident également à l’ADN de <i>D. ditremus</i>. Seules deux paires d’amorces, qui amplifient les loci Dd_8 et Dd_33, se sont avérées spécifiques à <i>D. dendriticus</i>. L’efficacité et la reproductibilité élevée des amorces Dd_8 ont été validées sur ~3 500 plérocercoïdes de <i>D. dendriticus</i> et de <i>D. ditremus</i> d’Islande et de Norvège. Ces amorces sont recommandées pour la différenciation moléculaire future des deux espèces de <i>Dibothriocephalus</i>. </p>

Review of African Political Economy (ROAPE)

<p>Since 1974 the Review of African Political Economy has provided radical analysis of trends, issues and social processes in Africa, adopting a broadly materialist interpretation of change.</p>

Drug Repurposing Research Collection

<p>Drug Repurposing Research</p>

Topical issue "Using wavefronts: detection and processing"

Uncertainty analysis of spectral flux measurement using Monte Carlo simulation | Journal of the European Optical Society-Rapid Publications

JEOS-RP: Rapid progress in optics and photonics has broadened its application enormously into many branches, including information and communication technology, ...