不同饲料饲育的家蚕幼虫肠道细菌的多样性分析

【目的】为研究饲料对不同家蚕Bombyx mori品种肠道微生物菌群的影响。【方法】以筛选到的家蚕广食性品种GS和普通品种1015为研究对象,收集从收蚁开始分别饲育桑叶(GS. m和C1015. m组)和人工饲料(GS. b组)至4龄盛时期的家蚕肠道样本,采用高通量测序的方法对其肠道微生物16S r DNA的V3-V4区进行测序分析,比较它们之间肠道微生物的差异。【结果】在门水平上,所测家蚕肠道样本的优势菌为厚壁菌门(Firmicutes)和变形杆菌门(Proteobacteria);在科水平上,所测样本主要优势菌为明串珠菌科(Leuconostocaceae)、乳酸杆菌科(Lactobacillaceae)、肠杆菌科(Enterobacteriaceae)等;在属水平上,所测样本主要的优势菌为魏斯氏属Weissella、乳酸菌属Lactobacillus、布赫纳氏菌属Buchnera、甲基杆菌属Methylobacterium、叶瘤菌属Phyllobacterium、肠球菌属Enterococcus和脆弱拟杆菌属Bacteroides等。家蚕品种GS经桑叶和人工饲料饲育后,甲基杆菌属Methylobacterium、布赫纳氏菌属Buchnera等菌属仅在桑叶饲育的GS肠道内出现,而魏斯氏菌Weissella、短芽孢杆菌属Brevibacillus等菌属只在人工饲料饲育的GS肠道内出现。同是桑叶饲育的家蚕品种GS和1015,其肠道内相同的优势菌有叶瘤菌属Phyllobacterium、脆弱拟杆菌属Bacteroides、不动细菌属Acinetobacter等。相较于广食性蚕品种GS的肠道菌群,肠球菌属Enterococcus、草螺菌属Herbaspirillum、丝硫菌属Thiothrix等菌属仅在普通蚕品种1015肠道中被检测到。GS. b组家蚕肠道细菌的物种多样性低于GS. m和C1015. m。GS. m肠道中丰度差异显著性最高的菌群为变形菌门(Proteobacteria),GS. b肠道中丰度差异显著性最高的菌群为杆菌纲(Bacilli)和乳杆菌目(Lactobacillales),而C1015. m肠道中丰度差异显著性最高的菌群为粪肠球菌属Enterococcus和肠球菌科(Enterococcaceae)。【结论】经桑叶饲育的不同蚕品种(GS和1015)的肠道微生物比人工饲料饲育的家蚕肠道微生物更趋于一致;经桑叶饲育的广食性家蚕肠道微生物物种多样性较高于经人工饲料饲育的广食性家蚕。     

饲料中转基因成分对家蚕生长发育的影响

采用家蚕（Bombyx mori）H9品种作为实验动物,分别用转基因和非转基因大豆粉制作的人工饲料进行饲育。通过饲育试验,比较分析了不同处理后家蚕的龄期经过时间、全茧量、茧层量、茧层率、蛹体重、疏毛率、死亡率、每龄眠起体重等经济性状指标。同时采用PCR方法对转基因饲料饲育后家蚕组织中外源基因的表达情况进行了检测。结果显示,含转基因大豆粉的人工饲料对家蚕的生长发育并无显著影响,且不存在外源基因的污染。     

不同饲料饲养家蚕其肠道微生态优势菌群类型的组成及差异性

为探讨鳞翅目昆虫的生长发育及抗病性与肠道微生态状况的关系,以不同的桑科植物柘叶与桑叶分别饲养家蚕,采用纯培养分离检测技术、16S rDNA序列测定和系统发育分析方法,对4、5龄家蚕肠道优势菌群的类型进行了鉴定和差异性分析。结果表明:柘叶与桑叶饲养家蚕共有的优势菌群有短波单胞杆菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)、肠杆菌属(Enterobacter)和葡萄球菌属(Staphylococcus)4个类群。从桑叶饲养家蚕肠道中检索到的优势菌群还有气单胞菌属(Aeromonas)、短杆菌属(Brevibacterium)、柠檬酸杆菌属(Citrobacter)、埃希氏菌属(Escherichia)和克雷伯氏菌属(Klebsiella)5个类群,而从柘叶饲养家蚕肠道中检索到的优势菌群仅有假单胞菌属(Pseudomonas)和土壤杆菌属(Agrobacterium)2个类群。饲料的改变导致家蚕肠道微生态细菌种群组成的变化,从柘叶饲养家蚕肠道中分离出的优势菌群与桑叶饲养的家蚕相比,出现较大差异且不如桑叶饲养家蚕的菌群丰富。推测这种改变可能与柘叶饲养家蚕生长发育不良、容易患病具有相关性。     

天蚕稚蚕期室内饲育方式的比较

本文通过天蚕1～2龄期6种饲育方式的比较,明确单头育和群育之间在1～2龄历期、死亡率、2龄末体重及以后各龄的历期、死亡率、结茧率、茧层量等方面均无显著差异。说明大规模饲育中稚蚕期可采取群育法。     

家蚕在人工饲料饲养下若干因素对生长的影响

为了探索家蚕人工饲料饲养实用化的途径,对人工饲料的配方、剂型以及幼虫的生活习性等进行了研究。结果表明:1.光照(全明或昼明夜暗)条件下人工饲料饲养的蚕发育较快而整齐,且增加了对CPV(质型多角体病毒)的抵抗力。2.下述配方较好:桑叶粉45克,豆饼粉25克,淀粉5克,石油酵母5克,蜂蜜5克,柠檬酸3克,维生素C2克,没食子酸0.4克,山梨酸0.3克,氯霉素25毫克,成型剂石花菜15克,加水250毫升。3.配方中桑叶粉含量必须保持在40%以上,质量以春叶为优。4.与桑叶饲育比较,蚕对营养成分的吸收率高,但转化率低。5.为了便于贮藏和运输,人工饲料可制成干片。     

一种家蚕嗜食的草本饲料植物

1)生菜是一种家蚕嗜食性良好的草本饲料植物,在目前草本饲料中甚属少见,可以代替5龄期食桑量的1/3强。 2)纯食生菜的家蚕,会引起死亡,这可能是生菜中含有校多的生菜硷所引起的中毒,如用桑菜间隔饲育法及混合饲育法,完全可以消除这些现象,同时可完成家蚕生长发育的周期。 3)家蚕多食生菜,发育延迟,生命力弱,茧质下降,今后如用驯化方法培育生菜家蚕的饲育品种,可以发挥生菜的饲用价值。 4)生菜与桑叶的合理饲育方法,应推生菜与桑叶的简隔轮饲法最好,其间隔时间为12小时左右,与家蚕消化吸收的生理过程上,获得同化上的协调作用。     

两种饲料植物对天蚕饲育的效果(英文)

通过五个处理的一系列比较,白栎Quercus fabri叶片作为珍贵绢丝昆虫——天蚕Antheraea yamamai的饲料,无论幼虫存活率、化蛹率、二至四眠蚕体重、五龄幼虫体重增长曲线及理论极限体重K值、取食量、相对摄食速率、消化量、结茧率、全茧量、蛹重、茧层量等均明显高于过去普遍采用并认为理想的麻栎Q. acutissima的饲育结果,从而证实白栎是饲育天蚕的最佳饲料植物。     

家蚕血液过氧化氢酶活力及其与蚕体抗逆性的关系

家蚕召Bombyx mori(L.)血液过氧化氢酶(CAT)活力随发育时期呈现有规律的动态变化。在幼虫期,CAT活力以龄初、龄末较高,而龄中较低;化蛹后,CAT活力迅速上升,至中期达到峰值,随后迅速下降直至羽化。不同性别间CAT活力均以雄性较高;不同蚕品种间CAT活力有很大差异;饲料对CAT活力有一定影响,人工饲料育蚕CAT活力略大于桑叶育的;高温冲击和氟化物添食都能引起CAT活力的显著变化,但变化的幅度因品种、性别、时期和添食剂量而有所不同。研究认为,家蚕血液CAT活力与其发育变态、体内代谢均具有密切关系,逆境条件下CAT活力变化幅度的大小可以作为家蚕抗逆性强弱的一个重要生理指标。     

大螟人工饲养技术的初步研究

利用大豆粉、玉米粉、麦胚和鲜茭白等成分配制了大螟Sesamia inferens(Walker)的半合成人工饲料,利用该饲料配方,发展了大螟的长期、继代饲养技术,即初孵-2日龄幼虫在茭白上饲养,3日龄-化蛹在人工饲料上饲养。利用该方法连续饲养大螟3代,幼虫的发育历期、蛹重、幼虫存活率、化蛹率、羽化率、卵孵化率和单雌产卵量等生活史参数与在天然饲料茭白上饲养的大螟相比,二者没有任何显著差异。而且利用该方法饲养,成本低、省工省力,能显著减少病原菌的感染。这说明该饲养技术适于大螟种群的长期、继代饲养。     

人工饲料、转Bt水稻及其亲本稻苗饲养下二化螟种群适合度的比较研究

以人工饲料、转Bt水稻"克螟稻"(cry1Ab纯和基因型)及其对照亲本"秀水11"稻苗为供试寄主植物开展二化螟Chilo suppressalis(Walker)1～5龄幼虫的室内饲养试验,以明确不同龄期二化螟种群的生活史参数。试验结果表明:二化螟在低龄时死亡率最高。克螟稻对二化螟各个龄期表现出高抗性,其各个龄期在克螟稻上均不能化蛹,随着龄期的增加二化螟的耐受性增强。以秀水11和人工饲料饲养二化螟对其蛹期、成虫期、单雌产卵量、羽化率的影响无显著性差异,以人工饲料饲养的二化螟蛹重显著高于以秀水11饲养的二化螟的蛹重,蛹重与人工饲料饲养时间呈正相关。与秀水11幼苗相比,人工饲料饲养下有利于二化螟雌虫的分化。     

Role of specific activators of intestinal amino acid transport in Bombyx mori larval growth and nutrition.

Nutrient absorption and its modulation are critical for animal growth. In this paper, we demonstrate that leucine methyl ester (Leu-OMe) can greatly increase the activity of the transport system responsible for the absorption of most essential amino acids in the larval midgut of the silkworm Bombyx mori. We investigated leucine uptake activation by Leu-OMe in brush border membrane vesicles and in the apical membrane of epithelial cells in the midgut incubated in vitro. Moreover, the addition of this strong activator of amino acid absorption to diet significantly affected larval growth. Silkworms fed on artificial diet supplemented with Leu-OMe reached maximum body weight 12-18 h before control larvae, and produced cocoon shells up to 20% heavier than those of controls. The activation of amino acid absorption plays an essential role in larval development so that larval growth and cocoon production similar to controls reared on an artificial diet with 25% of dry mulberry leaf powder were observed in silkworms fed on an artificial diet with only 5% of mulberry powder. Arch.     

Effect of different artificial diets on diapause induction under controlled temperature and photoperiod in the silkworm, Bombyx mori L.

ABSTRACT. Embryonic diapause of the silkworm, Bombyx mori , is generally induced by temperature and photoperiod during the egg stage of the previous generation and not in the larval stage. However, when silkworm larvae are reared on an artificial diet instead of mulberry leaves, their diapause is strongly affected by temperature and photoperiod experienced in the larval stage, with a distinct long-day response for diapause induction. Moreover when larvae which have been reared on artificial diet under long-day condition are fed mulberry leaves even for a short period of time, most of the resultant female adults lay diapause eggs. These results suggest that the photoperiodic response of larvae for diapause induction may be strongly suppressed by some components in mulberry leaves.     

Biological, nutritional, and histochemical basis for improving an artificial diet for Bracon hebetor Say (Hymenoptera: Braconidae)

The biology of Bracon hebetor Say (Hymenoptera: Braconidae) reared on fifth instars of Anagasta kuehniella (Zeller) (Lepidoptera: Pyralidae) (natural diet) and in vitro (artificial diet) was evaluated. Data on the number of instars, development time and food intake were collected, and histochemical tests were conducted to detect proteins and lipids in the parasitoid's digestive tract. The data disclosed differences that can help to improve artificial rearing of B. hebetor. B. hebetor had three instars in both diets, but the developmental time on the artificial diets was prolonged due to the increase in larval and pupal development times. Larvae grew faster on the natural host and required a lower food intake (2.7 microl) as compared to that required by the larvae feeding on the artificial diet (3.8 microl). Analysis of diet protein content and host hemolymph and the observations on the parasitoid larvae gut content indicated altogether the artificial diets requires the addition of others sources of proteins and lipids to improve the overall nutrition quality of the in vitro rearing system for this ectoparasitoid.     

Performance of polyvoltine silkworm Bombyx mori L. on different mulberry plant varieties

Five cultivated mulberry plant varieties, BSRM‐34, BSRM‐56, BSRM‐57, BSRM‐58 and BSRM‐59, were used to evaluate the effects of plant variety on larval and cocoon characteristics of mulberry silkworm Bombyx mori L. Silkworm larvae undergo their entire larval periods on leaves of mulberry varieties. Different larval characteristics of larval weight, larval length and larval breadth were measured in different growth stages. After rearing, the fifth instar larvae were transferred into a mountage (Chandraki) structure for cocoon rearing for spinning and cocoon production. The cocoons were then harvested and different cocoon characteristics like cocoon weight, shell weight and shell percentage; and other characters like pupal weight and effective rate of rearing were measured. The results showed statistically significant differences among different varieties on larval and cocoon characteristics. The highest performance was observed by feeding the variety BSRM‐34 in respect of the average weight of 10 mature larvae (40.5 g), single mature larval length (7.6 cm) and breadth (1.0 cm), single cocoon weight (1.4 g), shell weight (0.2 g) and pupal weight (1.2 g) while the average performance was recorded by feeding varieties BSRM‐57 and BSRM‐59. The poorest performance was showed by feeding the variety BSRM‐56 (in case of larval characters) and BSRM‐58 (in case of cocoon characteristics). These two varieties also showed the poorest performance for silk yield. Through comparing all the above parameters it was concluded that BSRM‐34 is the best mulberry variety and it is recommended that Bangladeshi farmers adopt that variety for better yield of silk.     

Rearing the 28-spotted ladybird beetle,Henosepilachna vigintioctopunctata (Coleoptera: Coccinelidae), with a switchover from host plant leaves to artificial diet

In this study, I tested an artificial diet, Insecta LFS, for rearing the 28-spotted ladybird beetle, Henosepilachna vigintioctopunctata (Fabricius). H. vigintioctopunctata larvae could not be reared through all of the larval stages on the artificial diet. However, initially they could be reared on tomato leaves up to the second or third instar, and thereafter exclusively on the artificial diet. The larval and pupal periods were not significantly different from those reared only on tomato leaves. For females reared by the diet-switching method, the preoviposition period was significantly longer and the number of eggs laid significantly lower than for those on tomato leaves. However, these differences did not become a hindrance to laboratory rearing. This rearing method saves labor involving obtaining fresh plant leaves and produces a reliable supply of the insects.     

人工饲料与天然饲料饲养条件下烟青虫的生长发育和繁殖力比较

报道饲养烟青虫HelicoverpaassultaGue澖ee的一种人工饲料,比较了烟青虫取食该饲料和天然饲料的主要生物学指标,测定了6龄幼虫对2种食料的转化和利用率。该人工饲料的主要成分是麦胚、黄豆粉、酵母粉和烟叶粉,天然饲料为烟草叶片。结果表明:取食人工饲料的烟青虫幼虫生长发育快,化的蛹较大,成虫寿命长,产卵量大,所产卵孵化率高;取食人工饲料的烟青虫与取食烟草蒴果的相比,前者相对取食量和近似消化率较高,相对生长率无显著性差异。说明烟青虫对该人工饲料有较好的适应性,可以作为繁殖烟青虫的实用饲料。     

New larval agar-based diet for laboratory rearing of Mediterranean fruit fly <Emphasis Type="Italic">Ceratitis capitata</Emphasis> (Diptera,Tephritidae)

Five larval diets for laboratory rearing of Ceratitis capitata were tested. These diets were based on wheat bran, microcellulose, potato starch and agar. To evaluate the quality of diet, pupal rearing efficacy and pupal weight were checked. The best results were obtained with an agar based diet used for Manduca sexta laboratory rearing. To simplify the preparation and to reduce the cost of this diet, a new formulation was developed. Larvae reared on the new agar-based diet achieved higher pupal rearing efficacy than larvae reared on bran diet recently used in medfly mass rearing facilities. Heat treated medflies reared on the new agar-based diet achieved similar pupal rearing efficacy with heat treated medflies reared on bran diet. When testing population density, higher pupal rearing efficacy was again achieved on new diet. The highest pupal rearing efficacy was achieved with 100 eggs per 25 g of diet, lowest with 500 eggs per 25 g of diet. Concerning pupal weight, there was no difference in results achieved on Petri dishes with different larval population densities. Larvae reared on new agar-based diet reached better results than larvae reared on bran diet. The preparation of the new diet is simple and the cost is low, so it is good for laboratory tests and rearing.     

Recycling of urea associated with the host plant urease in the silkworm larvae,Bombyx mori

Urea concentration and urease activity in the midgut content were compared between larvae of the silkworm, Bombyx mori fed an artificial diet and those fed fresh mulberry leaves. A considerable amount of urea was found in the midgut content of the both larvae, however it was significantly lower in the larvae fed fresh mulberry leaves than in the larvae fed the artificial diet; average urea concentrations in the midgut content of the larvae fed fresh mulberry leaves and the artificial diet were 2.9 and 4.6 &mgr;mol/g, respectively. Urea in the midgut content seems to be secreted from the insect itself since the amount of urea in both diets were negligibly small. Urease activity was detected only in the midgut content of the larvae fed fresh mulberry leaves but not in other tissues of the larvae. On the other hand, no urease activity was detected in the midgut content of the larvae fed the artificial diet. Subsequently, to elucidate the role of mulberry leaf urease in the midgut lumen, larvae that had been reared on the artificial diet were switched to fresh mulberry leaves. The diet switch caused a rapid decrease in urea concentration in the midgut content and an increase in ammonia concentration in the midgut content, suggesting that secreted urea could be hydrolyzed to ammonia by mulberry leaf urease in the midgut lumen. Furthermore, to investigate the physiological significance of mulberry leaf urease on urea metabolism of the silkworm, (15)N-urea was injected into the hemocoel, and after 12 h the larvae were dissected for (15)N analysis. A considerable amount of (15)N was found to be incorporated into the silk-protein of the larvae fed fresh mulberry leaves, but there was little incorporation of (15)N into the silk-protein of the larvae fed the artificial diet. These data indicate that urea is converted into ammonia by the action of mulberry leaf urease in the midgut lumen and used as a nitrogen source in larvae fed mulberry leaves.