美洲大蠊水解氨基酸含量测定及营养评价

采用酸水解法,测定美洲大蠊中氨基酸的种类和含量,并对其进行营养评价。结果表明:美洲大蠊含有17种氨基酸(色氨酸被水解破坏而未检出),总氨基酸质量分数为43.14%,其中药效氨基酸占总量的55.43%,呈味氨基酸占总量的20.28%,其氨基酸配比较为合理:E/(E+N)=0.35,E/N=0.55,与WHO/FAO提出的参考蛋白模式接近。美洲大蠊中氨基酸种类齐全,含量丰富,富含必需氨基酸和药效氨基酸,具有很高的食用、药用价值,有较好的开发潜力。     

四种蜚蠊的呼吸信号特征比较

在20℃下,利用CO₂红外分析仪采集了美洲大蠊Periplaneta americana、褐斑大蠊P.brunnea、澳洲大蠊P.australasiae和德国小蠊Blattella germanica的呼吸信号。结果表明,它们均具有典型的不连续气体交换循环（discontinuous gas exchange cycle, DGC）呼吸模式,且一个DGC可分为爆发间期和爆发期2个阶段。4种蜚蠊雄性成虫DGC的特征各异：美洲大蠊完成一个DGC周期平均约需24.55 min,明显长于褐斑大蠊(11.67 min)和澳洲大蠊(10.75 min),德国小蠊周期最短,仅为4.41 min;对于爆发间期历时在整个DGC历时所占的比例,美洲大蠊最大,平均为57％,德国小蠊次之,为48％,褐斑大蠊和澳洲大蠊比较接近,分别为37％和36％。德国小蠊单位体重的CO₂平均释放速率最大,而另外3种蜚蠊的差异不明显。4种蜚蠊爆发期CO₂释放体积均随DGC历时增加而增加,美洲大蠊和德国小蠊单位体重的CO₂平均释放率随DGC历时增加而减少,在褐斑大蠊和澳洲大蠊中它们关系不明显。美洲大蠊、褐斑大蠊和澳洲大蠊CO₂平均释放率和爆发期CO₂释放体积与体重呈正相关,在德国小蠊中关系不明显;4种蜚蠊DGC各阶段历时和DGC频率与它们体重的关系均不明显。     

美洲大蠊成虫肠道菌群多样性分析

美洲大蠊Periplaneta americana既是卫生害虫,也是重要的药用昆虫。许多研究表明,其肠道菌群种类和数量与宿主健康密切相关。本实验利用16S r RNA高通量测序技术分析了人工饲养的美洲大蠊成虫肠道菌群。结果得到有效序列67 492条,归类为581个操作分类单元,注释到21门79目193属。厚壁菌门Firmicutes(56.96%)和拟杆菌门Bacteroidetes(33.13%)为美洲大蠊成虫肠道的优势菌门。葡萄球菌属Staphylococcus(16.59%)、芽孢杆菌属Bacillus(13.26%)和副杆菌属Parabacteroides(10.26%)为美洲大蠊成虫肠道的优势菌属。研究结果为进一步探讨美洲大蠊肠道菌群多样性以及与宿主之间的关系打下了基础。     

三叶虫茶营养成分的分析与评价

通过与老鹰茶虫酿茶和一些品牌绿茶的对比,对三叶虫茶的一般营养成分、茶的生化特征、氨基酸和脂肪酸组成、维生素、矿物质元素进行分析和营养评价,为开发三叶虫茶提供科学依据。方法:用常规方法分析其主要营养素和生化成分;用氨基酸分析仪分析氨基酸组成,并采用WHO推荐的蛋白质模式对蛋白质进行营养评价;维生素和矿物元素含量直接说明其营养功能。三叶虫茶含有36.44%的水浸出物,16.28%的茶多酚、1.39%的氨基酸;氨基酸的种类较为齐全,总含量与传统茶叶相当,9种人体必需氨基酸的总量达到0.722%,是传统茶叶的3~12倍,特别是赖氨酸、蛋氨酸、苏氨酸、色氨酸的含量远远高于常规茶叶,而这4种氨基酸在一般植物食品中都是限制性氨基酸;但其茶氨酸、天门冬氨酸、谷氨酸的含量远远低于常规茶叶,而这3种氨基酸对改善茶汤滋味具有重要作用;三叶虫茶中脂肪酸含量为1.23%,其中多不饱和脂肪酸占35.25%;钙、磷、镁含量较高,铁、铜、锌、锰等的含量丰富;维生素C和维生素E含量与普通茶叶相当。三叶虫茶营养丰富,具备常规茶叶的一些生化特征,可作为茶饮料的代用品;若与普通茶叶混合使用,能在滋味和营养价值上实现互补。     

美洲大蠊microRNA的初步研究

美洲大蠊Periplaneta americana是一种世界性的卫生害虫,但其提取物有较高的药用价值。本研究对饲养的雌雄美洲大蠊成虫进行microRNA测序,分别在雄性和雌性中得到12 155 616条和9 847 263条序列。序列长度主要为18～23 nt,且在22 nt和29 nt处有2个峰值。将所得序列与数据库(NCBI、Rfam)进行比对注释,最终在雄性成虫中鉴定到57种已知的microRNA和152种潜在的新microRNA,在雌性成虫中鉴定到53种已知的microRNA和94种潜在的新microRNA。差异表达分析发现只有一种microRNA:miR-750在雌雄之间差异表达,其在雌虫中表达量显著高于雄虫。本研究首次在基因组水平研究了美洲大蠊microRNA的组成并对其功能进行了预测,为其后续研究奠定了基础。     

美洲大蠊成虫肠道抗细菌共生放线菌的筛选及鉴定

【目的】鉴于野外美洲大蠊Periplaneta americana对恶劣环境适应性强,本研究旨在从云南省大理州的野外美洲大蠊成虫肠道中分离、筛选出抗细菌活性放线菌,为抗生素开发提供菌种资源。【方法】采用涂布平板法和平板划线法对美洲大蠊成虫肠道放线菌进行分离;以金黄色葡萄球菌Staphylococcus aureus、耐甲氧西林金黄色葡萄球菌、铜绿假单胞菌Pseudomonas aeruginosa、粪肠球菌Enterococcus faecalis、大肠杆菌Escherichia coli和鼠伤寒沙门氏菌Salmonella typhimurium共6种人体病原细菌为指示菌株,采用牛津杯法对分离自这些放线菌的次生代谢产物进行抗菌活性测定;通过形态学特征和16S rRNA基因序列分析,对具有广谱和明显抗菌活性的放线菌进行鉴定,并经16SrRNA基因序列的BlAST同源性比对及系统发育分析确定它们的分类地位。【结果】从美洲大蠊成虫肠道共分离获得41株放线菌。抗菌活性测定结果表明,34株(82.9%)放线菌对至少1种指示病原细菌具有抑制作用,其中有7株对3种以上病原细菌具有抑制作用,9株表现出明显的抗菌活性。根据系统发育分析结果,这些放线菌均被鉴定为链霉菌属Streptomyces spp.。【结论】野外美洲大蠊成虫肠道含有丰富的抗细菌活性的放线菌资源,为后续挖掘新型抗生素提供重要的微生物资源。     

美洲大蠊胸腺素THY3的表达模式分析

美洲大蠊Periplaneta americana胸腺素基因具有THY1、THY2和THY3三个不同剪接体,其中THY3结构域最多。本研究使用实时荧光定量PCR技术比较分析了THY3在美洲大蠊不同性别、不同发育阶段及不同组织中的表达差异,以及大肠杆菌Escherichia coli诱导对美洲大蠊血淋巴和脂肪体中THY3表达的影响。结果表明:THY3在成虫期的表达量显著高于其他虫期,雌虫表达量显著高于雄虫,脂肪体表达量显著高于血淋巴、头部、肌肉、体壁组织。雌性成虫体腔注射大肠杆菌3 h后血淋巴中THY3表达量显著增高,而在脂肪体中12 h后才检测到THY3表达明显上调,研究结果为进一步研究美洲大蠊胸腺素的功能打下了基础。     

美洲大蠊粪便粗提物的聚集活性

用不同溶剂不同方法提取美洲大蠊Periplaneta americana(L.)粪便,测定美洲大蠊各虫态的诱集活性。结果表明,采用直接浸泡提取方法,丙酮、乙醇、正己烷和二氯甲烷4种粗提物对美洲大蠊各虫态都具有明显的诱集作用,其中乙醇和丙酮粗提物的引诱效果最好,正己烷次之,二氯甲烷最弱。4种溶剂粗提物对美洲大蠊雄成虫和高龄若虫聚集活性最强,对低龄若虫聚集活性最弱。用乙醇溶剂对粪便粗提,3种提取方法均对美洲大蠊有很强的诱集效果,其中索氏抽提诱集效果最弱,直接浸泡和超声波提取效果好,且差异不显著,但直接浸泡提取效果更好。     

杀虫环对昆虫血淋巴糖和游离氨基酸的影响

 用毛细管色谱和离子交换色谱法分析了黑胸大蠊雄性成虫血淋巴糖和游离氨基酸的组分、合量及杀虫环对它们的影响,并与电生理记录的EPSP电位比较,表明:1.黑胸大蠊血淋巴有五种糖,海藻糖、葡萄糖含量最多。2×10~(-5)mol/L杀虫环作用于虫体5分钟后,五种糖含量迅速增加;30分钟后下降23～47％。2.血淋巴中含有16种游离氨基酸及大量的NH_3。杀虫环可降低14种氨基酸的含量,尤其对生糖氨基酸(丙、异亮、酪、脯)的影响显著。3.2×10~(-5)mol/L杀虫环作用5分钟可增加中枢第Ⅵ腹神经节自发电位的释放频率和振幅,6～7分钟后迅速减少并消失,此时诱发EPSP电位振幅下降。18～20分钟突触传递被阻断。此过程恰巧与血淋巴糖代谢的加快而减缓的过程一致。     

美洲大蠊基因组重复序列分析

重复序列是真核生物基因组的重要组成部分。一些重复序列,如自主型的逆转录转座子LINE,在昆虫的系统进化和遗传多样性研究方面得到了广泛的应用。de novo从头预测和基于同源比对预测相结合的方法被用来搜索美洲大蠊Periplaneta americana基因组,共鉴定出大约占全基因组62%的重复序列。研究发现,散在重复序列中,DNA转座子占美洲大蠊基因组的16.18%;逆转座元件中LINE最多,占基因组的13.64%,SINE和LTR分别占基因组的3.52%和1.32%。LINEs中的Bov Bs亚家族在所有转座子亚家族中比例最高(约6.73%)。美洲大蠊与德国小蠊Blattella germanica相比,除LTR外,其他类型的转座子占基因组的比例均高于德国小蠊。通过分析逆转录转座子反转录酶完整度、氨基酸序列相似度及遗传距离,从美洲大蠊基因组中鉴定出一类BovBs:RTE-1_PAm。BovBs的反转录酶氨基酸序列的系统树表明,美洲大蠊与内华达古白蚁Zootermopsis nevadensis的进化关系比与其同属蜚蠊科Blattidae的德国小蠊的关系更近。昆虫中BovBs的进化关系与传统核基因进化关系的不同,表明转座子的进化相对宿主基因的进化具有一定的独立性。     

Cataract formation following limited amino acid intake during gestation and lactation

Female rats were fed defined diets limiting in one or more of certain amino acids and with or without vitamin E throughout gestation and lactation. Deficits of tryptophan, phenylalanine/tyrosine, or methionine/cystine reduced the body weight of progeny to about 50% or less of normal but only low tryptophan was cataractogenic. When total dietary amino acids were 12.4%, a low (65 mg%) level of tryptophan resulted in 34% incidence of cataract if vitamin E was simultaneously withheld. Elevation of total amino acids to 24.8% while maintaining tryptophan at 65 mg% caused 70 or 90% incidence of nuclear lens opacities in the presence or absence, respectively, of vitamin E. Maternal dietary amino acid imbalance was also associated with a 50% decrease in lens insoluble (membrane) proteins in the progeny independent of dietary vitamin E or the occurrence of opacities.     

PVY侵染后烟草营养成分的变化及其对介体烟蚜生长发育的影响

【目的】明确马铃薯Y病毒(potato virus Y,PVY)侵染后诱导的烟草营养成分的变化及其对烟蚜Myzus persicae生命特性的影响,旨在进一步解析PVY-烟草-烟蚜三者间的互作机制。【方法】通过蒽酮比色法和氨基酸自动分析仪测定了PVY不同侵染时期烟株体内的可溶性糖和游离氨基酸含量的变化;测定和比较了感病与健康烟草植株上烟蚜种群生长发育、成虫寿命、繁殖力和有翅蚜产生量的差异性。【结果】PVY侵染前、中、后期(分别为侵染后5,12和20 d)的烟草叶片中游离氨基酸的总量均显著高于健康烟草叶片。相较于健康烟草叶片,在PVY侵染前期的烟草叶片中,谷氨酸、脯氨酸、天冬氨酸、色氨酸、缬氨酸、赖氨酸和组氨酸的含量显著增加;PVY侵染中期,感病叶片中丝氨酸含量显著下降,谷氨酸、天冬氨酸、色氨酸、缬氨酸、亮氨酸、苯丙氨酸、精氨酸和组氨酸含量显著提高;PVY侵染后期,感病叶片中甘氨酸含量显著下降,谷氨酸、脯氨酸、天冬氨酸、苏氨酸、缬氨酸、亮氨酸、丙氨酸、苯丙氨酸、组氨酸、酪氨酸和精氨酸含量显著提高。在PVY侵染的前期和中期,感病叶片中的可溶性糖含量显著高于健康烟叶,而在侵染后期感病叶片中可溶性糖含量显著低于健康烟草叶片的。PVY侵染前期和中期的烟草叶片中总糖和总游离氨基酸的含量比值显著高于健康烟草叶片中的。在PVY侵染的烟草植株和健康烟草植株上取食的烟蚜其发育历期、若蚜历期、成蚜繁殖期、繁殖后期、寿命、烟蚜种群的内禀增长率、周限增长率和平均世代周期均无显著差异,但在感病烟草植株上取食的烟蚜成蚜繁殖前期显著缩短,其繁殖力和净生殖率显著提高。相较于健康烟草植株,在PVY侵染烟草植株上定殖的烟蚜种群有翅蚜发生的高峰期提前。【结论】PVY侵染前期和中期提高了寄主烟草的营养品质,从而提高了烟蚜的繁殖力。侵染后期烟草营养品质的下降,促使烟蚜种群有翅蚜的产生和扩散,从而有利于PVY自身的传播。     

Aspects of amino acid metabolism and nutrition in the ectoparasitoid wasp, Exeristes roborator

Exeristes roborator contains a wide variety of free amino acids, and the composition of all developmental stages was quantitatively dominated by proline and glutamic acid. The latter occurred together with lesser amounts of glycine, alanine, valine, leucine, tyrosine, and histidine which varied between developmental stages. Minor and trace amounts of most other commonly occurring amino acids were also found. The percentages or relative amounts of major constituents were not influenced when the parasite was reared on the alternate hosts, Pectinophora gossypiella and Gnorimoschema operculella. Likewise, the major characteristics of the latter hosts' free amino compositions could not be accounted for, in either case, by their diets.Differences in the relative distribution of the major amino acids between the developmental stages of E. roborator indicate large decreases in the percentage of proline occur during development from the larval to the adult stage with corresponding increases in the percentages of glycine in the pupal stage and alanine as well as other amino acids in the adult stage. The results suggest that proline may play an important rôle in E. roborator, probably as an energy reserve.The amino acid compositions of the total proteins of E. roborator and its hosts were similar and all quantitatively dominated by glutamic acid, aspartic acid, and amides. However, the disk gel electropherograms of the proteins of both hosts and parasite were different. Quantitative changes were evident in the protein pattern of the parasite when reared on the alternate hosts.E. roborator incorporated radioactivity from ¹⁴C(U)-glucose into the amino acids, glutamic acid, aspartic acid, serine, glycine, alanine, and proline. Furthermore, ¹⁴C(U)-glutamic acid was incorporated into a wide variety of proteins. The data suggest that the above amino acids may be non-essential dietary components for E. roborator. However, quantitative determinations indicate that the amount of glutamic acid synthesized does not account for the amount incorporated into protein over the same time period.     

Regulation of Growth and Gametangial Formation in Riccia gangetica Ahmad by Some Amino Acids

The present work deals with the effect of six amino acids: asparagine,aspartic acid, glutamic acid, glycine, serine, and tryptophan,on growth and gametangial formation in Riccia gangetica. Allthe amino acids tested enhance vegetative growth, and amongthese glutamic acid proves best. The total number of rhizoidsis reduced in response to amino acids. Aspartic acid and glutamicacid favour antheridial production. In contrast, asparagine,serine, and tryptophan enhance archegonial formation, and amongthese asparagine elicits the best response. Glycine proves bestfor antheridial production, and also increases the number ofarchegonia. Key words: Riccia gangetica, Amino acids, Growth, Gametangial formation     

Nutritional Composition and Assessment of Gracilaria Iemaneiformis Bory

The chemical composition, mineral elements, vitamins, free fatty acids and amino acid content of the edible red alga Gracilaria lemaneiformis Bory, grown in the sea near Nan'ao island, Guangdong Province, were analyzed in the present study. Gracilaria lemaneiformis Bory showed a total sugar content of 14.65%. The protein content was 21%, of which approximately 41% was determined to be essential amino acids (EAA).The major amino acid components were glutamic acid, leucine, arginine, and alanine. Of the EAA assayed,methionine and cysteine appeared to be the most limiting amino acids compared with the EAA pattern provided by Food and Agricultural Organization of the United Nations. The total lipids content was 0.87% and comprised a high composition of unsaturated fatty acids (61%), mainly as linoleic acid and oleic acid, and a little amount of polyunsaturated fatty acid; palmitic acid was the main component (39%) of saturated acids.Relatively high levels of vitamin C, iodine, phosphorus, and zinc were also present in G. lemaneiformis. The nutritional composition between G. Iemaneiformis and Nostoc flagelliforme, a rare alga that is widely eaten in Chinese society, was compared. The results suggest that N. flagelliforme can be substituted for by G.Iemaneiformis, not only because of their similar shape, but also because of their approximate nutritional composition. Gracilaria Iemaneiformis may possibly serve as a potential healthy food in human diets in the future.     

Quantitative analysis of free amino acids and carbohydrates in spring barley anthers at different stages of maturity

The content of the carbohydrates glucose, fructose and sucrose was determined in spring barley anthers at different stages of maturity. During maturation the sucrose content of the anthers increased markedly. The following 17 free amino acids were detected in anthers of different stages of maturity: aspartic acid, glutamic acid, serine, alanine, arginine, leucine, isoleucine, lysine, α-aminobutyric acid, glutamine, proline, tyrosine, phenylalanine, valine, threonine, cystine and glycine. Quantitative analysis was only carried out in amino acids present in higher concentrations in the analysed samples. These were: aspartic acid, glutamic acid, α-aminobutyric acid, proline, serine, valine and glutamine, and a mixture of amino acids (leucine, isoleucine, valine and phenylalanine). The total content of free amino acids increased with increasing maturity of the anthers. However, not all amino acids followed contributed to this increase, but only proline, glutamic acid, aspartic acid and glutamine. A small difference was found in the variety Gopal in which the aspartic acid content did not increase significantly, but the content of the mixture of amino acids and serine did. With the exception of green anthers of the variety Firlbecks Union, proline was present in the highest concentration in all samples analysed.     

组培苗鲜罗汉果与干罗汉果营养成分的分析和比较

对组培苗鲜罗汉果与干罗汉果提取物的营养成分进行分析和比较研究,分别测定了蛋白质、脂肪、总糖、碳水化合物、灰分、矿质元素、维生素E和氨基酸含量。结果表明:组培苗鲜罗汉果和干罗汉果蛋白质含量分别为22.23%、19.8%,氨基酸总量分别为12.51%、8.36%,其中人体必需氨基酸占氨基酸总量分别为38.13%、36.36%,矿质元素含量分别为1081mg/100g、1089mg/100g,维生素E含量分别为0.183mg/100g、0.589mg/100g。组培苗鲜罗汉果的蛋白质、氨基酸含量比组培苗干罗汉果分别高2.43%、4.15%,碳水化合物、维生素E含量组培苗干罗汉果比组培苗鲜罗汉果分别高2.44%、0.406mg/100g,矿质元素、粗脂肪、总糖、灰分等含量两者不存在明显差异。     

Isolation and analysis of natural compounds from silkworm pupae and effect of its extracts on alcohol detoxification

Silkworm pupae have much potential and many applications as a natural medicine to promote human health. However, their chemical components have not been fully characterized or understood. HPLC analysis was conducted to determine the content ratio (%) of individual amino acids in total protein of the pupae. It showed that glutamic acid (18.3%), histidine (14.6%) and alanine (10.2%) are the most common amino acids in silkworm pupae. Fatty acid composition of silkworm pupae oil was revealed by high‐pressure liquid chromatography and gas chromatography – mass spectroscopy analyses. They contain a high ratio of essential fatty acids, [α‐linolenic acid (ω‐3 fatty acid]+ linoleic acid) (49.0%), and also contain non‐essential fatty acids, oleic acid (19.9%), palmitoleic acid (2.5%), palmitic acid (19.7%), stearic acid (8.6%), and eicosapentaenoic acid (EPA) (0.3%). In addition, they also contain antioxidants, quercetin diglucoside and nutritionally important riboflavin (vitamin B2). This study suggests that silkworm pupae are a nutritionally valuable food product and are applicable as cosmetic components with essential amino acids, essential fatty acids, antioxidants and vitamins. The animal experiment showed that alcohol dehydrogenase (ADH) activity was significantly higher in the liver of mice orally administered with 0.5 mg/mL of silkworm extract and alcohol than with commercial Dawn808? and alcohol, indicating that silkworm pupae extracts have alcohol detoxification activity.