黑水虻幼虫和预蛹的饲料价值研究进展

黑水虻Hermetia illucens L.是一种全世界广泛分布的资源昆虫,幼虫在自然界以动物粪便、腐烂的有机物,如腐肉、腐烂的水果、蔬菜和植物性垃圾为食,能够被用于畜禽粪便的无害化处理。黑水虻幼虫在将粪便转化成昆虫生物量的同时,减少了粪便的污染。利用畜禽粪便饲养的黑水虻幼虫干重达到42%～43%,其中粗蛋白含量占干重的42%～44%,脂肪含量占干重的31%～35%,灰份占干重的11%～15%,钙质占干重的4.8%～5.1%,磷占干重的0.60%～0.63%。使用粪便饲养的幼虫和预蛹,含有丰富的必需氨基酸和矿物质,是家禽、家畜和鱼类养殖的良好饲料来源。文章综述利用不同动物粪便饲养的黑水虻幼虫和预蛹的饲料价值及其幼虫和预蛹用于鸡、猪和鱼类饲料添加剂的研究进展。     

黑水虻对小龙虾废弃物转化效果研究

我国水产养殖业近年来发展迅速,同时也增加了大量水产废弃物,小龙虾Procambarus clarkii废弃物就是其中之一。黑水虻Hermetia illucens能将有机废弃物转化成高附加值的昆虫蛋白,因此,本研究将小龙虾废弃物与蛋白含量较高的豆腐渣混合,设计6个不同混合比例的饲料,筛选出高转化率和高性价比的饲料配方,分析饲料中营养物质降解情况和预蛹的营养成分。结果表明：小龙虾废弃物与豆腐渣按照4∶1混合,黑水虻存活率较高（93.42%±2.65%）,幼虫产量最高（44.27±1.30 g）,料重比最低（2.26±0.07）,预蛹粗蛋白含量可达43.93%±0.49%;饲料残渣中的粗蛋白和粗脂肪含量比原饲料均有降低,其中粗脂肪降幅最大,说明黑水虻对脂肪利用率很高。综合评价认为小龙虾废弃物与豆腐渣4∶1混合为本实验范围内最佳饲料配方,本研究为进一步利用黑水虻处理小龙虾废弃物奠定了基础。     

Effects of food substrates and moxidectin on development of black soldier fly,Hermetia illucens

Black soldier fly is a common and widely distributed saprophagous species that has an excellent potential for being used for biological conversion of organic wastes on an industrial scale. The main goal of the reported study was expanding the list of wastes suitable for utilization by this species. We compared larval growth on cull potatoes, horse manure and cafeteria food waste in 100‐L bins in a greenhouse. We also conducted laboratory experiments to investigate whether black soldier fly larvae are affected by the presence of moxidectin, a common endectocide used to treat an array of domestic animals and readily excreted in faeces, in their food substrates. Feeding on potatoes resulted in slower growth, and the final size of potato‐fed larvae was smaller compared to the larvae fed on cafeteria waste. Nevertheless, potatoes supported substantial biomass accumulation, and could be a valuable option for rearing fly larvae for commercial feed production. Larvae feeding on horse manure gained very little weight and eventually failed to pupate. Moxidectin had a strong negative effect on larval survivorship; however, ca. 30% of larvae reared in the substrate containing a realistic field concentration of moxidectin still survived to adulthood. Our findings confirm that using black soldier fly larvae is a promising technology for recycling organic wastes, including those of plant origin.     

亮斑扁角水虻对有机废弃物的应用及其肠道微生物研究进展

有机废弃物的安全处理与二次利用是当今社会发展进程中的一大难题,亮斑扁角水虻Hermetia illucens强大的生物转化能力可有效解决此类问题：一方面,亮斑扁角水虻可大量消耗有机废弃物,有效治理环境污染;另一方面,转化后的虫体营养价值高,广泛应用于养殖领域。然而,亮斑扁角水虻的生物转化作用机制目前尚不明确,可转化的有机废弃物范围及其转化效率的影响因素仍有待探讨。本文结合近年来关于亮斑扁角水虻领域的研究,主要从亮斑扁角水虻对有机废弃物的生物转化作用、转化效率的影响因素、虫沙的应用和肠道微生物的功能4个方面进行综述,同时对未来需要解决的问题进行了展望,以期为亮斑扁角水虻的综合开发利用提供参考依据。     

Bioconversion of poultry manure and vegetable waste mixes by Hermetia illucens (Diptera: Stratiomyidae)

Development time, survival and final nutrient content of Hermetia illucens L. larvae depends on the substrates in which they develop. Mixing different waste types together can increase the performance and survival of the larvae, as well as their waste reduction. The main objective of this study was to evaluate the effect of different ratios of mixed fruit and vegetable waste with poultry manure on larval development time, size, biomass production, survival, bioconversion and waste reduction. Freshly hatched neonates (90 mg; approx. 6000 individuals) were placed on 12 kg of a mixture of fruit and vegetable waste and fresh, unprocessed poultry manure and held at 28 ± 0.5°C. Inclusion of fruit and vegetable waste varied from 0% to 100% in 10% increments. Initial temperature of the substrate was also measured. The individual mass of larvae increased significantly as more fruit and vegetable waste was included, from less than 81.3 ± 6.6 mg on poultry manure only to an average size of 211.6 ± 6.0 mg at 100% fruit and vegetable waste. After approximately 60% inclusion of fruit and vegetable waste the performance and survival of the larvae increased significantly while development time was reduced. A combination of high fruit and vegetable waste and low initial temperatures resulted in lower development time overall. The mixing of wastes can be applied in industry to further the goals of waste reduction and biomass production while incorporating low-quality wastes like poultry manure.     

Development,growth and metabolic rate of Hermetia illucens larvae

The larvae of Hermetia illucens are known to successfully bio‐convert a vast range of organic substrates into high protein and fat biomass, but little is known about the larval instars. During this research, larval head capsules and biomass growth were measured daily and the specific metabolic rate of larger instars were considered. The head capsule measurements revealed that H. illucens pass through 6 actively feeding larval stadia before entering the last nonfeeding but migrating 7th stadium. Larval growth follows a sigmoid curve with slowly accelerating growth in the earlier stadia and decelerating growth in the latest stadia. In contrast, development was fast until reaching stadium 6 and then slowed down. Accordingly, the specific metabolic rate was high in instars 3, 4 and 5 and reduced in instars 6 and 7.     

黑水虻幼虫氨基酸及其微量元素分析

黑水虻具有较高的营养价值,是一种重要的环境友好型资源昆虫.本研究采用凯氏定氮法、茚三酮柱后衍生离子交换色谱法和微波消解-电感耦合等离子质谱法(ICP-MS)等多种分析技术,测定了厨余垃圾饲喂的黑水虻幼虫蛋白质、氨基酸和微量元素的组成及含量.研究结果表明,黑水虻幼虫中蛋白质的含量为40.60 g/100 g.在测定的16...     

Isolation and characterization of high-mobility-group proteins from maize

Chromosomal nonhistone high-mobility-group (HMG) proteins were purified from nuclei of maize (Zea mays L. cv. A619) endosperm and leaf tissue. Tissuespecific differences were observed in their polypeptide patterns, in in-vitro phosphorylation experiments with a casein-kinase type II, and by Western blot analysis with antisera against different HMG proteins. Gelfiltration chromatography demonstrated that maize HMG proteins occur as monomers. By measuring the capacity of the HMG proteins to bind to the 5 flanking region of a zein gene, the sensitivity of the proteins to different temperatures, salt concentrations and pH values was determined.Abbreviations EMSA electrophoretic-mobility-shift assay - FPLC fast protein liquid chromatography - HMG high-mobility group - kDa kilodaltons - PVDF polyvinylidenedifluoride - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis We would like to thank Mrs. E. Brutzer for excellent technical assistance. We are indebted to Mrs. M. Strecker and Dr. W. Bessler of the Institut für Immunbiologie, Freiburg, FRG, for the preparation of antisera and we gratefully acknowledge helpful discussions with Drs. T. Quayle, R. Grimm and U. Müller of this institute. This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fond der Chemischen Industrie.     

Effect of the rearing diet on gene expression of antimicrobial peptides in Hermetia illucens (Diptera: Stratiomyidae)

Insect proteins have been proposed for human and animal food production. Safeguarding the health status of insects in mass rearing allows to obtain high-quality products and to avoid severe economic losses due to entomopathogens. Therefore, new strategies for preserving insect health must be implemented. Modulation of the insect immune system through the diet is one such strategy. We evaluated gene expression of two antimicrobial peptides (one defensin and one cecropin) in Hermetia illucens (L.) (Diptera: Stratiomyidae) reared on different diets. Analyses were performed on prepupae and 10-day-old larvae reared on cereal- and municipal organic waste-based diets and on only prepupae reared on a cereal-based diet supplemented with sunflower, corn, or soybean oil. The inclusion of sunflower oil at different points in the cereal-based diet was also evaluated. Moreover, diet-driven differences in the inhibitory activity of the hemolymph were tested against Escherichia coli DH5α and Micrococcus yunnanensis HI55 using diffusion assays in solid media. Results showed that a municipal organic waste-based diet produced a significant overexpression of antimicrobial peptides only in prepupae. Inclusion of vegetable oils caused an upregulation of at least one peptide, except for the corn oil. Higher expression of both genes was observed when sunflower oil was added 5 days before pupation. All hemolymph samples showed an inhibitory activity against bacteria colonies. Our results suggest that municipal organic waste-based diet and vegetable oil-added diet may successfully impact the immune system of H. illucens. Such alternatives may also exist for other species of economic interest.     

不同废弃物对黑水虻幼虫生长、营养组成及生产性能影响研究

本文以黑水虻Hermetia illucens为研究对象,探索不同废弃物对其幼虫生长、营养组成及生产性能的影响。结果表明,黑水虻幼虫可将餐厨垃圾（FW）、厨余垃圾（KW）、蔬菜废弃物（VW）、猪粪（PM）、牛粪（DM）、鸡粪（CM）等转化成优质蛋白质和脂肪。以FW和KW作为基质养殖10 d后,幼虫体长（19.0 mm、17.1 mm）与单头重（171.54 mg、136.82 mg）显著高于其它处理。虫粉粗蛋白含量受废弃物种类影响相对较小。不同废弃物处理虫粉均显示出良好氨基酸谱。尽管各处理虫粉总必需氨基酸含量低于鱼粉和豆粕,但FW、KW和CM处理虫粉缬氨酸含量高于鱼粉和豆粕,VW处理虫粉异亮氨酸和缬氨酸含量高于鱼粉和豆粕,PM处理虫粉组氨酸和缬氨酸含量高于鱼粉和豆粕。以FW和KW作为基质养殖10 d后,尽管虫粉不饱和脂肪酸和必需脂肪酸比例低于采食PM和DM的处理,但远高于其它处理的虫粉粗脂肪含量（42.13%、37.49%）,虫粉不饱和脂肪酸和必需脂肪酸含量也最高;料虫比（6.06、7.90）显著低于其它处理,幼虫转化率（59.40%、52.07%）显著高于其它处理;废弃物减量化率（89.53%、84.46%）仅低于VW处理（94.86%）。利用黑水虻幼虫处理FW和KW,幼虫生长性能和营养组成优于其它废弃物处理,且生产性能最好。     

黑水虻抗菌肽HI-3对RAW264.7细胞免疫调控作用研究

为了明确黑水虻Hermetia illucensL.抗菌肽HI-3对RAW264.7细胞的免疫调控作用,拓展黑水虻抗菌肽在免疫领域的应用潜能,本文研究了抗菌肽HI-3对RAW264.7细胞形态、增殖、吞噬功能、分泌NO和细胞因子以及抗氧化活性的影响。结果表明,20、40、80、160、320μg/mL的抗菌肽HI-3对RAW264.7细胞形态及增殖指数均无显著影响,但均能显著增强RAW264.7细胞的吞噬功能,且随着HI-3浓度增加吞噬能力逐渐增强,当HI-3浓度达到320μg/mL时RAW264.7细胞的吞噬能力最强(P<0.01);而在160μg/mL以下处理浓度时,对NO含量无显著性影响(P>0.05);抗菌肽HI-3可以上调RAW264.7细胞分泌IL-6、TNF-α和IL-1β,下调抑炎因子IL-10的分泌量,且具有一定的浓度依赖性,浓度达到320μg/mL时效果最为显著(P<0.01);另外,抗菌肽HI-3亦能够显著提高RAW264.7细胞超氧化物歧化酶(SOD)活力和总抗氧化能力(T-AOC)含量,尤其是320μg/mL时,效果最为明显(P<0.0...     

响应面法优化黑水虻幼虫蛋白质提取工艺

【目的】对黑水虻Hermetia illucens幼虫蛋白质进行不同提取方法的比较,选择最优提取方法,并确定其最优工艺参数,为黑水虻幼虫蛋白提取与资源利用提供依据。【方法】以黑水虻幼虫为原料,分别采用碱提法、酶提法、盐提法和Tris-HCl缓冲液提法对黑水虻幼虫蛋白质进行提取,并比较分析。通过单因素试验分别确定NaOH质量浓度、液料比、提取温度及提取时间4个因素的较优水平。在单因素试验结果基础上,按照Box-Behnken响应面试验设计进行响应面优化试验。【结果】提取黑水虻幼虫蛋白质的4种方法中碱提法的提取率最高,最佳提取条件为：NaOH质量浓度2.44 g/100mL,液料比22 mL/g,提取温度53℃,提取时间2 h。提取率验证试验结果为88.49%,与预测值相对误差为0.28%。【结论】响应面模型拟合度高,优化出的最佳提取工艺可行。     

工业蛋白质理性设计与应用

作为合成生物学与绿色生物制造等领域的底层核心技术,蛋白理性设计可有效解决天然功能元件性能不足等共性挑战,创制高性能人工酶元件。值此天津工业生物研究所(Tianjin Institute of Industrial Biotechnology, TIB)创立10周年之际,文中回顾了研究所在工业蛋白理性设计领域的系列重要工作进展。从酶设计方法学研究、新酶反应设计到生物催化应用等方面进行了分析讨论,并展望了本领域未来发展方向。望借此搭建学术界和产业界与酶理性设计的桥梁,促进新技术、新策略的开发应用,加速融合人工酶的基础研究与产业应用,推动我国生物制造领域的科技创新升级。     

黑水虻消化道形态及组织结构的观察

本文比较了不同发育阶段黑水虻Hermetia illucens消化道的形态学差异,掌握了幼虫消化系统的组织学特征。利用体视镜观察黑水虻5龄幼虫、预蛹及成虫的消化道形态,利用光学显微镜和扫描电镜观察幼虫消化道各段(前肠、中肠、后肠)的显微及超微结构。结果表明：黑水虻幼虫及预蛹的消化道均由前肠(食道和前胃)、中肠及后肠组成,从幼虫到成虫,消化道的长度不断缩短。与幼虫和预蛹相比,成虫消化道形态变化明显,前胃消失,出现了嗉囊及胃盲囊,中肠进一步缩短,后肠分化为回肠、结肠和直肠。组织学观察结果显示,幼虫的唾液腺开口于口腔,由膨大的管状腺体和腺管组成。食道由特化为角质刺突的内膜层及发达的肌层组成,其末端延伸至前胃。前胃膨大为球状,包括三层组织结构。根据上皮细胞形态的差异,中肠可分为四个区段。后肠薄,肠腔内褶丰富,肠壁可见数量较多的杆状细菌。马氏管开口于中、后肠交界处,包括4支盲管,管内壁密布微绒毛。黑水虻消化道形态随发育阶段的变化,反映了各阶段摄食及消化生理的差异。幼虫消化道各段具有各自典型的组织学特征,其前、中、后肠可能分别承担了食物接纳与初步消化、消化与吸收以及重吸收功能。本研究结果为进一步了...     

Modeling bond correlations in denatured proteins and polypeptides

Bond‐orientational correlations for finite‐length homopolypeptides and a selected group of denatured proteins are obtained by numerical simulations using a polypeptide model with a potential of mean force. These correlations characterize the stiffness of the polypeptide backbone and are generally described by either an exponential or a power‐law decay in the asymptotic limit. However, for finite length polypeptides and unfolded proteins the correlations significantly deviate from either single exponential or power‐law behavior. A heuristic model is developed to analyze the correlations of homopolypeptides, which depends on the chain length and the side‐chain properties. The model contains power‐law and multi‐exponential terms, the latter which could be interpreted as local persistence lengths. In the asymptotic limit, the model reduces to the expected power‐law behavior. Simulations of denatured proteins show that the power‐law behavior of the correlations is significantly suppressed and only the multi‐exponential term is needed to model the correlations. In addition, average persistence lengths (ranging from 2.0 to 2.5 nm) are obtained from the correlations by fitting single exponentials and shown to be in general agreement with experiments, which also assume single exponential decay. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 312–323, 2016.     

黑水虻幼虫蛋白及其酶解产物的抗氧化活性研究

为了探究黑水虻Hermetia illucens幼虫蛋白及酶解产物的抗氧化活性,以鲜活黑水虻幼虫冻虫为原料,采用碱提酸沉法提取黑水虻蛋白,通过碱性蛋白酶、菠萝蛋白酶、风味蛋白酶、木瓜蛋白酶对其蛋白质溶液进行酶解,分别从2,2′-联氮-双-3-乙基苯并噻唑啉-6-磺酸（ABTS）自由基、羟自由基、1,1-二苯基-2-三硝基苯肼（DPPH）自由基3个方面对黑水虻蛋白及其酶解液的抗氧化能力进行测定。结果表明：黑水虻幼虫蛋白及4种酶解后的蛋白肽具有较好的抗氧化活性,其中黑水虻幼虫蛋白对ABTS自由基、羟自由基、DPPH自由基的半抑制浓度（IC50）分别为2.91、0.232、0.764 mg/mL,而酶解过后的蛋白肽具有更强的抗氧化活性,对ABTS自由基、羟自由基、DPPH自由基的最低半抑制浓度（IC50）分别为0.295、0.082、0.354 mg/mL。本研究初步证明了酶解制备抗氧化肽的可行性,为昆虫蛋白资源的利用和无抗饲料的研制提供了新的研究思路。     

Genetic and phenotypic consequences of early domestication in black soldier flies (Hermetia illucens)

The black soldier fly, Hermetia illucens, is an emerging biotechnological agent with its larvae being effective converters of organic waste into usable bio-products including protein and lipids. To date, most operations use unimproved commercial populations produced by mass rearing, without cognisance of specific breeding strategies. The genetic and phenotypic consequences of these commercial practices remain unknown and could have a significant impact on long-term population viability and productivity. The aim of this study was thus to assess the genetic and phenotypic changes during the early phases of colony establishment and domestication in the black soldier fly. An experimental colony was established from wild founder flies and a new microsatellite marker panel was developed to assess population genetic parameters along with the phenotypic characteristics of each generational cohort under captive breeding. The experimental colony was characterised by a small effective population size, subsequent loss of genetic diversity and rapid genetic and phenotypic differentiation between the generational cohorts. Ultimately, the population collapsed by the fifth generation, most likely owing to the adverse effect of inbreeding depression following the fixation of deleterious alleles. Species with r-selected life history characteristics (e.g. short life-span, high fecundity and low larval survival) are known to pose particular challenges for genetic management. The current study suggests that sufficient genetic and phenotypic variations exist in the wild population and that domestication and strain development could be achieved with careful population augmentation and selection during the early stages of colony establishment.