取食聚苯乙烯塑料对黄粉虫幼虫肠道微生物群落的影响北大核心

随着塑料在人类社会中的普及,越来越多的废弃塑料及其前体物质被遗留在环境中,且其在自然环境中的降解速度十分缓慢,为此寻找有效的降解途径成为亟待解决的科学问题。【目的】探究利用黄粉虫(Tenebrio molitor)幼虫取食聚苯乙烯对其肠道微生物种群及其代谢路径的响应,以期通过食物诱导寻找一条生物降解和利用聚苯乙烯的有效途径。【方法】以聚苯乙烯为唯一食物来源喂饲黄粉虫幼虫,通过测量幼虫存活率和个体的体重来测定其生长发育情况;通过对其肠道内容物进行16S rRNA基因测序,分析其肠道菌群结构的变化;采用京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析法来预测相关功能基因。【结果】取食聚苯乙烯黄粉虫幼虫存活率和体重均下降,聚苯乙烯塑料明显减少;取食聚苯乙烯的黄粉虫幼虫肠道菌群丰度与多样性明显减少,在门水平,取食聚苯乙烯的黄粉虫幼虫肠道的优势菌为变形菌门(Proteobacteria)、软壁菌门(Tenericutes)和厚壁菌门(Firmicutes);在属水平,取食聚苯乙烯的黄粉虫幼虫肠道优势菌为螺旋体菌属(Spiroplasma)、肠杆菌(Enterobacillus)和大肠埃氏-志贺氏菌(Escherichia-Shigella);通过KEGG功能预测,找到与芳香类和烷烃降解功能相关的基因共18种,取食聚苯乙烯组黄粉虫幼虫肠道菌群降解聚苯乙烯相关通路丰度升高,相关基因表达增强。【结论】聚苯乙烯可以为黄粉虫幼虫生长发育提供一定的物质和能量,且能够使其完成一个世代过程;幼虫长时间取食单一食物后,其肠道菌群结构会发生目标性变化,利用KEGG预测能够找到与聚苯乙烯代谢相关的基因,为后续研究工作提供了有价值的依据。     

大麦虫Zophobas morio对塑料的取食和降解作用研究

研究了大麦虫Zophobas morio对不同塑料制品的喜食性,并对大麦虫取食不同塑料原料制品后的排泄物进行了热重分析同步差热分析和傅里叶转换红外光谱分析.试验结果确定大麦虫取食聚氯乙烯后的排泄物与聚氯乙烯相比,红外特征峰以及热特性均发生较大的变化.     

大麦虫及其肠道微生物对聚乙烯的生物降解

【背景】废旧塑料聚乙烯因具有较高的化学惰性,不易被自然降解而形成长期污染。【目的】探究聚乙烯泡沫塑料对大麦虫生长发育的影响,为大麦虫作为降解聚乙烯泡沫塑料的昆虫推广提供理论依据。【方法】以大麦虫幼虫为研究对象,选用常见的泡沫塑料(聚乙烯),采用4种不同的饲喂方式T1 (麦麸)、T2 (泡沫塑料)、T3 (泡沫塑料+麦麸)、T4 (不饲喂)进行驯化,处理30 d后对大麦虫进行解剖,取肠道内容物于LB培养基中进行富集培养,将富集培养后的菌液加入以聚乙烯(polyethylene,PE)为唯一碳源的LCFBM培养基进行选择性培养,从中筛选分离得到对PE塑料有降解能力的菌株。【结果】取食泡沫塑料30d后,与单一饲喂PE相比,麦麸和PE混合饲喂后大麦虫幼虫的存活率为76%。采用傅里叶变换红外光谱检测发现虫粪组分中主要官能团中峰值明显变化,表明PE长链有断裂现象,并从肠道中分离得到3株可以对PE薄膜边缘造成明显侵蚀的菌株。【结论】大麦虫可取食并消化PE塑料,其肠道内的微生物对PE塑料的降解起到关键作用,研究结果为塑料污染的生物降解提供了科学证据。     

聚乙烯塑料的微生物降解

聚乙烯(polyethylene,PE)是产量最大的通用塑料之一,通常被加工成一次性包装材料(包括塑料袋及容器)和农用薄膜等。PE塑料的广泛应用导致大量PE废弃物的累积,对生态环境造成严重的威胁。自20世纪70年代以来,一些研究陆续报道了PE塑料被微生物降解的现象,并从土壤、海洋、垃圾堆置点及昆虫肠道等生境中分离筛选到了若干种具有一定PE塑料降解能力的菌株,而且发现一些单加氧酶、过氧化物酶和漆酶等氧化还原酶对PE塑料具有氧化降解能力。这些研究为发展PE塑料废弃物生物降解处理技术提供了一定的依据。本文总结和分析了PE塑料降解微生物的分离和筛选方法,以及已报道的PE塑料降解微生物和降解酶的研究进展,以期为进一步研究PE塑料的微生物降解机理和处理技术提供参考。     

黄粉虫对不同比例聚苯乙烯的取食效率研究

为探索黄粉虫Tenebrio molitor在降解聚苯乙烯过程中聚苯乙烯与麸皮的最佳配比,本研究将聚苯乙烯与麸皮按不同比例PS1(1:9),PS2(2:8),PS3(3:7),PS4(4:6),PS5(5:5),PS6(6:4),PS7(7:3),PS8(8:2),PS9(9:1),PS10(10:0),CK(0:10...     

聚乙烯塑料生物降解研究进展

聚乙烯(polyethylene,PE)塑料是全球通用合成树脂中产量最丰富的品种,也是最难降解的塑料之一,其在环境中大量积累已造成严重的生态污染。传统的垃圾填埋、堆肥和焚烧处理技术难以满足生态环境的保护要求,生物降解是解决塑料污染问题的一种生态友好、成本低廉、前景可期的方法。本文对PE塑料的化学结构、降解微生物的种类、降解酶和代谢途径等方面进行了综述,结合国内外PE塑料生物降解的前沿和热点问题,建议重点开展高效降解菌株筛选、人工合成菌群构建、降解酶的挖掘与改造等方面的研究,为PE塑料生物降解研究提供路径选择和理论借鉴。     

黑粉虫幼虫肠道细菌的研究

从黑粉虫幼虫肠道环境中分离、纯化、培养,获得5个细菌菌株,对其培养性状、染色反应、菌体形态、生理生化反应等进行了系统研究.鉴定结果表明：上述5个细菌菌株分别属于金杆菌属（Aureobacterium）、李斯特菌属（Listeria）、微杆菌属（Microbacterium）、莫拉菌属（Moraxella）、短小杆菌属（Curtobacterium）,其中金杆菌属、微杆菌属、短小杆菌属菌也发现于黄粉虫幼虫肠道环境中.通过研究,既可为黑粉虫生产所需微生态制剂提供微生态理论依据,又发掘了微生物资源,丰富了微生物资源库.     

解塑再用-中欧合作项目“合成塑料降解转化微生物菌群”介绍

中欧环境生物技术合作研究重大项目废弃塑料的生物降解由国家自然科学基金委员会(NationalNatural Science Foundation of China,NSFC)与欧盟委员会(European Commission,EC)共同资助,旨在促使中欧科学家在"Microorganism communities for plastics biodegradation"领域开展实质性合作研究。该项目的目标是将易造成环境污染且难以降解的石化塑料,利用微生物菌群的代谢能力降解为单体小分子并为微生物所用,从而进一步实现高值生物化学品的生物合成。这不仅可以解决塑料污染问题,同时也将塑料垃圾"变废为宝",创造更高的经济效益。中欧合作研究项目将促进双方科学家在合成生物学领域的深入合作,有助于双方建立长期稳定的国际交流与合作关系。中欧双方将致力于解决全球塑料污染问题,形成科技战略力量,共同开启探索不可降解塑料资源化利用领域的新篇章。     

桑粒肩天牛幼虫肠道菌群的研究

Intestinal flora of 47 Apriona germari(Hope) larvae,collected from fields,had been isolated and identified.The results showed that the predominant bacteria were Staphylococcus.Its viable count was 7.63±0.21,and the detection rate was 100%.Meanwhile,a strain of cellulose-utilizing bacterium was isolated from the fore-midgut fluid of A.germari larvae with the cellulose-congo red agar medium.The bacterium was tentatively identified as Cellulomonas.The detection rate of the cellulolytic bacterium was 23.40%,and…     

发展生物可降解塑料的途径和前景

塑料作为高分子聚合物的第三代材料应用于工业、农业,医药、国防以及人们日常生活等方面发挥重要作用,塑料制品处处司见,但这类人造的化学塑料制品不论以何种形式存在,一旦废弃于环境,则难以为生物降解。这种性能有其利也有其弊,作为一种“塑料垃圾”就必然给环境造成一大危害,如一些旅游地区及海洋等水域都可见到这些塑料废弃物。为此,从     

Prostaglandin biosynthesis by fat body from larvae of the beetle Zophobas atratus

We describe prostaglandin (PG) biosynthesis by microsomal-enriched fractions of fat body prepared from larvae of the tenebrionid beetle, Zophobas atratus. PG biosynthesis was sensitive to incubation time, temperature, pH, substrate and protein concentration. Optimal PG biosynthesis conditions of those we examined included 2 mg of microsomal-enriched protein, incubated at 22 degrees C for 2 min at pH 6. These preparations yielded four major PGs: PGA(2), PGE(2), PGD(2) and PGF(2 alpha). PGA(2) and PGF(2 alpha) were the predominant eicosanoids produced under these conditions. Two non-steroidal anti-inflammatory drugs, indomethacin and naproxen, effectively inhibited PG biosynthesis in low concentrations. In vitro PG biosynthetic reaction conditions, using vertebrate or invertebrate enzyme sources, usually include a cocktail of reaction co-factors. The Z. atratus preparation similarly performs better in the presence of co-factors. Arch.     

Cardioactive properties of Solanaceae plant extracts and pure glycoalkaloids on Zophobas atratus

Glycoalkaloids, the biologically active secondary metabolites produced by Solanaceae plants, are natural defenses against animals, insects and fungi. In this paper, the effects of glycoalkaloids present in extracts of Solanaceae plants (potato, tomato and black nightshade) or pure commercial glycoalkaloids on the coleopteran Zophobas atratus F. were evaluated by in vitro and in vivo bioassays using heart experimental models. Each tested extract induced a dose‐dependent cardioinhibitory effect. The perfusion of Zophobas atratus semi‐isolated heart using the highest potato and tomato extract concentration (1 mmol/L) caused irreversible cardiac arrests, while extract from black nightshade produced fast but reversible arrests. Pure commercial glycoalkaloids caused similar but less evident effects compared with extracts. Our results showed that the bioactivity of tested compounds depended on their structure and suggested the existence of synergistic interactions when combinations of the main glycoalkaloids of potato and black nightshade were used for trials. Surprisingly, injection of tomato and potato extracts in 1‐day‐old pupae of Zophobas atratus induced reversible positive chronotropic effects and decreased the duration of the both phases (anterograde and retrograde) of the heart contractile activity. Furthermore, these extracts affected the amplitude of the heart contractions.     

Ecdysteroid titres in a tenebrionid beetle,Zophobas atratus: effects of grouping and isolation

Metamorphosis in Zophobas atratus is dependent on isolation: when kept in grouped conditions, larvae undergo numerous supernumerary moults, growing in size, without pupating. This beetle thus represents an interesting model for the analysis of possible differences in the endocrine regulation of normal vs. supernumerary larval moults. In this study, the ecdysteroid titres have been analysed in this species, using enzyme immunoassay. The hormonal variations of larvae undergoing normal or supernumerary larval cycles were particularly examined, in either grouped or isolated conditions. Normal larval cycles presented very similar ecdysteroid variations in grouped as well as isolated conditions, showing a single hormonal peak (at about 1000pg/&mgr;l). Supplementary larval cycles, occurring in grouped conditions, also showed a similar single ecdysteroid peak, but after a longer period of basal levels. Isolation of such larvae triggered their larval-pupal transformation, which was characterized by more complex hormonal fluctuations, including a small ecdysteroid peak before the main one. Interestingly, the isolation of big larvae during a large part of their cycle induced this peculiar hormonal pattern synchronously, confirming the involvement of a complex neuroendocrine control between external conditions and ecdysteroid titres.     

Microscopic analysis of mechanosensory system monitoring the dynamic claw actions in the tenebrionid beetle Zophobas atratus

Many insects have a pair of claws on each leg. The distribution of mechanoreceptors that monitor claw actions was examined in the tenebrionid beetle Zophobas atratus. Each claw has 25–45 campaniform sensilla (CS) that detect the claw’s deformation due to substrate engagement. Five CS clusters are observed around the end of the 5th tarsomere (Ta5) in a concave, socket-like structure. The 1st cluster, containing 2–5 CS, is embedded in the unguifer to which the claws are articulated. The symmetrical 2nd and 3rd clusters, each containing two CS, are located bilaterally in the ventrolateral grooves of the sidewall of the socket, into which the unguis retractor plate slides. The 4th and 5th clusters, containing 1–2 CS with two hair sensilla, are localized near the ventrolateral ridges of the socket into which the basal portion of the claw is pressed during maximal claw flexion. In addition, Ta5 has a chordotonal organ of six sensory cells to monitor claw extension. These results suggest that the mechanoreceptor system may directly monitor the precise mechanical states of individual claws and provide the central nervous system with the sensory information required for fine feedback control of movements of the pretarsus and other leg segments for locomotion and other purposes.     

Molecular cloning,structural analysis,and the developmental expression of vitellogenin in the giant mealworm Zophobas atratus

The giant mealworm Zophobas atratus, which is used for commercial purposes, was officially introduced to Korea. However, physiological information of oocyte maturation for the mass rearing of Z. atratus has not been investigated. Here, we cloned Z. atratus vitellogenin (ZaVg) cDNA and analyzed the expression profiles during development to better understand oocyte maturation in Z. atratus. A multi-alignment and phylogenetic analysis using the deduced amino acids of ZaVg and those of other insect vitellogenins showed that ZaVg has typical Coleopteran Vg features. Consistent with ovarian development, lower expression levels of ZaVg protein in the fat body, hemolymph, and ovary were observed at 0 and 10 days in the adult stage, while higher expression levels of that were detected at 20, 30, and 40 days in the adult stage. These results suggest that the expression level of ZaVg is adult-stage specific, which providing basic information on Vg and oocyte development in Z. atratus.     

Structure-activity relationships for the cardiotropic action of the Led-NPF-I peptide in the beetles Tenebrio molitor and Zophobas atratus.

We have examined the effects of the Led-NPF-I peptide (Ala-Arg-Gly-Pro-Gln-Leu-Arg-Leu-Arg-Phe-amide) and a series of ten analogues on the heart contractile activity of Tenebrio molitor and Zophobas atratus, and the structure-activity relationships for cardioactive action of Led-NPF-I were established. A video microscopy technique and computer-based method of data acquisition and analysis were used to study the action of the peptides on continuously perfused heart preparations. Cardiac activity was progressively inhibited by Led-NPF-I when the peptide concentrations were increased from 10(-9) to 10(-5) M. Substitution of the L-proline residue at position 4 of the native peptide with hydroxyproline, valine or D-proline caused a loss of cardioinhibitory activity. Also, replacement of arginine residues at all three positions 2, 7 and 9 with another basic amino acid histidine, reduces cardioinhibitory action of Led-NPF-I. Some modifications of the C-terminal residues, as the Phe(4-NO2)-, Phe(4-NH2)- and Phe(4-NMe2)-analogues, resulted in agonistic peptides with biological activity similar to that of the native peptide. However, three other C-terminal analogues tested [Tyr10]-, [D-Phe10]-Led-NPF-I, and Ala-Arg-Gly-Pro-Gln-Leu-Arg-Leu-Arg-Phe-OH were inactive in the heart bioassay, which suggests that this end of the amino acid chain may play an important role in bioactivity and interaction of the native peptide with its receptor on the myocardium.     

Optimal hatching conditions of Zophobas atratus (Coleoptera: Tenebrionidae) eggs under various culture conditions

Supermealworm, Zophobas atratus (Coleoptera: Tenebrionidae) is a tropical species used for human consumption and as an ingredient in animal feed. We investigated the hatching characteristics of Z. atratus eggs under varying conditions to optimize artificial rearing conditions. Overall, the average egg weight, width, and length decreased by 42.3%, 3.7%, and 10.1%, respectively, on the 6th day post-oviposition. We varied either temperature, photoperiod, illuminance, or humidity, while the other conditions remained constant. Of the four experimental temperatures—25 °C, 27 °C, 30 °C, and 33 °C–30 °C was associated with the highest hatching rate (91.0%) during days 5–7. Although other experimental conditions did not have a significant effect on hatchability, we were able to elucidate the ideal conditions for improved hatchability. For photoperiod, hatchability was the highest (84%) at 12L/12D, followed by 81.0% at 15L/9D, and 70% at 9L/15D. Among the illuminance conditions tested—600, 1800, and 3000 lx—hatchability was the highest at 1800 lx (82.3% on days 4–6). Among the relative humidity (RH) levels tested—50%, 65%, and 80%—hatchability was the highest at 65% RH (88.5% on days 5–6). Thus, the ideal conditions for maximum hatchability were 30 °C, 12L/12D, 65% RH, and 1800 lx. Under these combined conditions, the hatching rate increased by 93.6%, and the hatching time was reduced by two days.     

Molecular identification and functional characterisation of uncoupling protein 4 in larva and pupa fat body mitochondria from the beetle Zophobas atratus

Uncoupling protein 4 (UCP4) is a member of the UCP subfamily that mediates mitochondrial uncoupling, and sequence alignment predicts the existence of UCP4 in several insects. The present study demonstrates the first molecular identification of a partial Zophobas atratus UCP4-coding sequence and the functional characterisation of ZaUCP4 in the mitochondria of larval and pupal fat bodies of the beetle. ZaUCP4 shows a high similarity to predicted insect UCP4 isoforms and known mammalian UCP4s, both at the nucleotide and amino acid sequence levels. Bioenergetic studies clearly demonstrate UCP function in mitochondria from larval and pupal fat bodies. In non-phosphorylating mitochondria, ZaUCP activity was stimulated by palmitic acid and inhibited by the purine nucleotide GTP. In phosphorylating mitochondria, ZaUCP4 activity decreased the yield of oxidative phosphorylation. ZaUCP4 was immunodetected with antibodies raised against human UCP4 as a single 36-kDa band. A lower expression of ZaUCP4 at the level of mRNA and protein and a decreased ZaUCP4 activity were observed in the Z. atratus pupal fat body compared with the larval fat body. The different expression patterns and activity of ZaUCP4 during the larval-pupal transformation indicates an important physiological role for UCP4 in insect fat body development and function during insect metamorphosis.