Enzymatic hydrolysis of silkworm pupa and its allergenicity evaluation by animal model with different immunization routes

Silkworm pupa is a nourishing food with high nutritional value,but its consumption has been greatly limited given its allergenicity.Enzyme hydrolytic technique is recognized as an effective method to reduce the allergenicity of protein.In this study,we aimed to investigate the effect of enzymolysis on the allergenicity of silkworm pupa.Crude silkworm pupa protein was extracted through alkali extraction and acid precipitation,which included 5 proteins with the molecular weights ranging from 34 kDa to 76 kDa,and silkworm pupa were then hydrolyzed by alkaline protease.The allergenicity of silkworm pupa protein and its enzymatic hydrolysates was evaluated by establishing BALB/c mice model,and the mice were immunized via intragastric gavage and intraperitoneal injection,respectively.The results indicated that the intraperitoneal inj ection immunization route induced more by detecting with antibodies,histamine and Th2-related cytokines.Moreover,mice treated with silkworm pupa protein peptide displayed no obvious allergic symptoms,indicating that enzyme hydrolytic technique could significantly reduce the allergenicity of silkworm pupa.

Effects of silkworm pupa protein on apoptosis and energy metabolism in human colon cancer DLD-1 cells

Objective:To investigate the effects of silkworm pupa(Bombyx mori) protein(SPP) on cell proliferation,apoptosis and energy metabolism in human colon cancer cells DLD-1.Methods:CCK-8 was used to detect cell proliferation rate after 72 h of cell culture for the control group(normal cultured DLD-1 cells) and SPP dose groups;Annexin-V/PI was applied to observe cell apoptosis;XFe24 Extracellular Flux Analyzer was used to detect cell mitochondrial respiratory function and glycolytic function.Results:Comparing with the control,SPP significantly inhibited the proliferation of DLD-1 cells with all the dosage tested(P <0.01);flow cytometry showed that SPP significantly promoted apoptosis(P<0.05).Additionally,SPP could significantly inhibited mitochondrial metabolism and glycolysis of DLD-1 cells and decreased cell energy metabolism in all groups treated with different doses.Conclusion:SPP can cause oxidative damage,promote apoptosis,and reduce mitochondrial respiratory and glycolysis rate in colon cancer DLD-1 cells,which reveals that SPP has the potential to serve as the anti-cancer drugs in the future,but further experimental evidence is needed.

Substitution of Silkworm Pupae （Bombyx mori） for Fish Meal in Broodstock Diets for Snakeskin Gourami （ Trichogaster pectoralis）

Substitution of silkworm pupae （Bombyx mori） for fish meal in broodstock diets for snakeskin gourami （Trichogaster pectoralis） was conducted in order to evaluate the effect of silkworm pupae on feed digestibility and broodstock performance. Snakeskin gourami broodfish were fed with isocaloric diets （DE 3,000 a： 50 Kcal/kg） contained 32% crude protein. The levels of substitution were 0%, 50%, 75% and 100% of protein from fish meal （equal to 0%, 14.57%, 21.75% and 29.03% silk worm by weight）. The study was assigned in CRD with 4 treatments and 3 replications. Six hundred of snakeskin gourami with five-month old and average weight of 78.67 ± 2.15 g in female and average weight of 73.00 ± 2.49 g in male were stocked in 12 cages. Fifty fishes of 25 females and 25 males were reared in each cage and fed experimental diet for five months. The result indicated that protein digestibility of test diets were significantly decrease （P 〈 0.05） according to the level of silkworm pupae in diets where as the dry matter digestibility showed no significantly different （P 〉 0.05）. The growth performance and egg fecundity were significantly decreased （P 〈 0.05） according to the levels of silkworm pupae. In contrast, egg quality in term of fry number on first spawned, fingering number on first spawned were not significantly different （P 〉 0.05）. The survival rate of one month nursing fish on first spawned were significantly different （P 〈 0.05）. Hence, silkworm pupae could substitution for 50% of protein from fish meal （14.57% by weight） in snakeskin gourami broodstock diets without any adverse effect on egg quality.

A Novel Protein Found in Selenium-rich Silkworm Pupas

A novel protein was isolated and characterized in selenium-rich silkworm pupas. The peptide mass fingerprint of the protein was found to be new. Partial amino acid sequencing also confn-med to be a new protein. The novel protein had a molecular mass of about 80 kDa in the SDS-PAGE.

Production Performance of Broiler Chickens Fed with Silkworm Pupa （Bombyx mon~

The production performance of broilers fed with silkworm pupa （Bombyx mori） was studied by focusing on growth, feed utilization, carcass composition and meat quality. There were two difference sources of silkworm pupa： （1） silkworm pupa from spun silk industry （SSP） and （2） silkworm pupa from silk yam reeling industry （RSP） were mixed into five isonitrogenous （20% CP） and isocaloric （ME = 3,150 kcal/kg feed） diet. The 195 chickens at three weeks old were equally distributed to 15 pens in the closed housing. The experiment was divided into five dietary treatments （three replications each）： treatment 1 （10% fishmeal, FM and 0% silkworm pupae） （control））, treatment 2 （0% FM and 10% SSP）, treatment 3 （0% FM and 20% SSP）, treatment 4 （0% FM and 5% SSP ＋ 5% RSP）, and treatment 5 （0% FM and 10% SSP ＋ 10% RSP）. Daily weight gain was 0.069, 0.060, 0.051, 0.065 and 0.061 kg/chick in treatments 1, 2, 3, 4 and 5, respectively. It is indicated that replacing fishmeal with silkworm pupa （5%SSP ＋ 5%RSP） performed the same daily weight gain as control but exhibited the higher feed conversion ratio （1.87） than control （1.68）. Carcass muscle of control treatment 1 （58.26%） was the highest （P 〈 0.05） and closed to treatment 4 and 5 （57.62% and 57.83%, respectively）. Sensory evaluation was not significantly differences （P 〉 0.05）. Therefore, treatment 4 （5%SSP ＋ 5%RSP） could be substituted FM with little adverse effect on broiler growth but without any adverse effect on percentage of muscle and sensory test. However, further supplemental silkworm pupa performed reduction in dietary utilization and carcass muscle.

Productive Performance of Giant Freshwater Prawn （Macrobrachium rosenbergi） Fed Diet Containing Silkworm Pupae （Bombyx mon~ Replacing for Fish Meal

The productive performance of giant freshwater prawn （Macrobrachium rosenbergii de Man） fed diet containing silkworm pupae （Bombyx mori） replacing for fish meal was conducted by focusing on digestibility, growth performance and feed utilization. Five diets of an isonitrogenous of 35 ：e 1% crude protein and isocaloric of digestible energy of 3,200 ±100 kcal/kg were formulated by using silkworm pupae, 0%, 8.6%, 17.2%, 25.8% and 34.7% by weight, replaced for fish meal. Giant freshwater prawn of 19 ± 2.0 g were stocked at 5 ind./m3 in concrete tank of 2.25 ton for three months. The protein digestibility by pepsin enzyme was evaluated in fish meal, silkworm pupae, soybean meal and shrimp meal. The digestibility was significantly difference （P 〈 0.05）. The protein digestibility of fishmeal and soybean meal was higher than silkworm pupae and shrimp meal. The productive performance of prawn in terms of growth rate and feed utilization were not significantly different （P 〉 0.05）. Therefore, silkworm pupae can be replaced for fishmeal in giant freshwater prawn （Macrobrachium rosenbergii de Man） without any adverse effects on productive performance.

Antihypertensive Properties on Spontaneously Hypertensive Rats of Peptide Hydrolysates from Silkworm Pupae Protein

Peptide hydrolysates of silkworm pupae protein with molecular weight of less than 5000 Da were prepared by ultrafiltration. The extracted peptide hydrolysates of silkworm pupae protein had inhibitory action on angiotensin-I-converting enzyme activity in vitro. The hydrolysates were orally administered to spontaneously hypertensive rats (SHR) in one period and long-term (four weeks). The results showed that the systolic blood pressure (SBP) of the treatment groups decreased in a dose-related manner. After one oral administration of silkworm protein hydrolysates with doses of 60, 20 and 5 mg/kg, the SBP of SHR decreased by 21.5, 13.8, and 9.0 mmHg in 1.5 h. After four weeks of the treatment in 80 mg/kg, the SBP decreased by 25 mmHg, with the antihypertensive activity close to 4 mg/kg of captopril;the SBP of the 40 mg/kg dose group also decreased by 17.5 mmHg. The peptide hydrolysate did not affect the SBP in normal, non-hypertensive rats in one period and long-term treatments. The acute toxicity research showed that the peptide hydrolysates were safe and without side effects. This research would be helpful in exploring the silkworm protein peptides as functional components for the antihypertension treatment.

Study on ionic liquids based novel method for separation and purification of silkworm pupa protein

Silkworm pupa protein(SPP)that obtained by traditional method usually had a high fat content,which would impose restrictions on the further use of SPP.Herein,various functionalized ionic liquids(ILs)were used to extract SPP from silkworm pupae,the structure-performance relationship of ILs with their SPP separation performance were explored at the same time.The research showed that the maximum extraction yield of SPP was up to 62.6%with less than 0.5%low fat content by using 1-ethyl-3-methylimidazolium chloride([Emim]Cl),when the dissolution experiment was conducted at 90°C for 24 h with ethanol bath as the regeneration solvent.Comparing with the structure of raw material,the regenerated SPP maintained the native protein backbone.Meanwhile,all regenerated SPP showed a decreased crystallinity,which also exhibited decreased fraction of theα-helix comparing to thatβ-sheet united with coil random structures.

Radon transform-based motion blurred silkworm pupa image restoration

As for machine vision-based intelligent system in the application of discriminating and sorting the sex of silkworm pupae,the tail gonad was the unique physiological feature.However,motion blur,resulting from the live silkworm pupa’s writhing motion at the moment of capturing image,could lose textures and structures(such as edge and tail gonad etc.)dramatically,which casted great challenges for sex identification.To increase the image quality and relieve the difficulty of discrimination caused by motion blur,an effective approach that including three stages was proposed in this work.In the image prediction stage,first sharp edges were acquired by using filtering techniques.Then the initial blur kernel was computed with Gaussian prior.The coarse version latent image was deconvoluted in the Fourier domain.In the kernel refinement stage,the Radon transform was applied to estimate the accurate kernel.In the final restoration step,a TV-L1 deconvolution model was carried out to obtain a better result.The experimental results showed that benefiting from the prediction step and kernel refinement step,the kernel was more accurate and the recovered image contained much more textures.It revealed that the proposed method was useful in removing the motion blur.Furthermore,the method could also be applied to other fields.

Comprehensive Investigation to Analyze the Amino Acid Constituents of the Spent Silk Worm Pupae of Anthereaea assama

Spent silk worm pupae ofAntheraea assama were analyzed for its amino acid constituents and compared with the amino acid composition of fiber from A. assama, A. pyrni, and Bombyx mori. Amino acid percent was determined by UV Spectrophotometer at 570 nm. Infra Red spectra recorded the presence of different amino acids in the spent silk worm pupae powder which corroborate with the amino acids present in the fiber of different silk species.

Distinguishing sex of silkworm chrysalis in cocoon by NIR with fiber optics

The feasibility of distinguishing male and female silkworm chrysalis in cocoon by NIR with fiber optics was discussed. Altogether 187 samples were taken from which 100 samples were used for calibration and others for prediction. "1"and"0"were designed as male and female silkworm chrysalis,respectively’,for regression. Statistical analysis results showed that MR was 0. 930 and SEC 0. 189. The prediction results of 87 samples indicated that R was 0. 937 and SEP 0. 175 respectively. The accuracy of distinguishing sex of silkworm chrysalis in cocoon reached 98. 9%. At the same time,it is possible to discriminate the dead silkworm pupa in cocoon immediately through the spectrum.

蚕蛹性腺图像在线采集与雌雄自动识别分选装置设计与优化

针对当前蚕蛹雌雄鉴别环节人工劳动强度大、蚕蛹雌雄识别分选装置准确率和效率不高、适用品种范围小等问题,根据雌雄蚕蛹性腺特征差异是区别不同品种蚕蛹雌雄的可靠依据,该研究设计了一种蚕蛹性腺图像在线采集与雌雄自动识别分选装置。首先,在对蚕蛹基本物理特性参数大量统计的基础上,对振动排序上料机、蚕蛹输送滑轨和蚕蛹翻转拍照装置进行了结构设计与理论分析。在识别方法和计算机硬件条件不变基础上,对装置中蚕蛹排序输送螺旋倾角、滑轨倾角和转动棒带动蚕蛹旋转转速进行了优化设计,以提高装置的识别分选速度。并设计了以SIEMENS-S7-200SMART-ST30为控制器,基于激光、光电传感器检测的信息融合控制系统。分选装置工作性能优化结果为:螺旋倾角为15.62°,滑轨倾角为31.77°,转速为21.09 r/min,单粒蚕蛹平均分选时间为5.13 s,准确率为95.97%,破损率为0.07%。样机试验结果表明,在最优参数下,利用卷积神经网络的全新轻量级雌雄识别分类模型,单粒蚕蛹平均分选时间为5.03 s,分选准确率为96.47%,破损率为0.09%,满足蚕蛹雌雄识别分选实际应用要求。该蚕蛹性腺图像在线采集与雌雄自动识别分选装置能够实现蚕蛹自动排序上料、蚕蛹带性腺图像在线采集、自动识别与雌雄自动分选的一体化,提高了蚕蛹雌雄自动识别分选的效率和准确率。研究结果可为雌雄蚕蛹自动识别分选装置的研发提供理论依据和技术支持。

家蚕雌蛹及其卵巢脂肪酸丰度分析

研究使用气相色谱分析了家蚕雌蛹蛹体和卵巢中的脂肪酸丰度.结果显示,蛹化后的第1 d至第6 d,蛹体脂肪酸丰度的变化幅度小,第6 d至第9 d,蛹体脂肪酸丰度的变化幅度大.蛹化期间,主要脂肪酸丰度总体呈下降趋势,微量脂肪酸丰度呈上升趋势.卵巢脂肪酸丰度的变化幅度随发育递减.不同主要脂肪酸的变化幅度和方向各异,其中不饱和脂肪酸丰度增加,饱和脂肪酸丰度下降.微量脂肪酸丰度随发育明显下降,这表明微量脂肪酸与雌性生殖关联较小.蛹体和卵巢脂肪酸丰度的差异随发育逐渐缩小,至蛹化第6 d,卵巢中绝大部分的脂肪酸丰度与刚蛹化时有很大差异,但整体上与蛹体丰度相似,说明卵巢脂肪酸的积累依赖蛹体脂肪酸.蛹化后期,卵巢中丰度最高的α-亚麻酸出现下降,丰度次高的油酸明显增加,这可能与卵壳生成和卵子成熟有关;亚油酸在蛹化第6 d丰度差异最大,其在卵巢中的丰度显著增加并明显高过蛹体,表明卵巢具有为其发育以及卵子发生积累脂肪酸的特性.研究结果可为深入研究脂肪酸在家蚕和其他昆虫雌性生殖中的作用机制提供重要线索.

基于纹理特征和改进VGG的家蚕蛹雌雄识别方法

【目的】针对蚕种培育中人工分蛹效率低且易受到主观因素影响的问题,提出一种基于纹理特征和改进VGG的家蚕Bombyx mori蛹雌雄识别方法。【方法】利用透射变换矫正蚕蛹方向,截取家蚕蛹头尾图,以B通道图作为轮廓提取的基础,通过道格拉斯-普克(Douglas-Peucker, DP)算法,分析轮廓复杂性从而识别并获取家蚕蛹尾部图;采取掩膜消除背景干扰,通过多通道的特征融合图加强纹理信息;对Inception模块进行改进,将残差网络与改进后的Inception模块加入VGG模型中;利用数据增强技术扩充数据集;以精确率(precision)、召回率(recall)、精确率和召回率的调和平均F1分值以及准确率(accuracy)作为评价指标,分别对3种输入图片以及4种识别模型进行评估对比。【结果】结果表明,特征融合图在改进VGG模型上的家蚕雌蛹的精确率、召回率和F1分值分别为98.017%, 94.794%和96.375%,雄蛹的精确率、召回率和F1分值分别为95.342%, 98.231%和96.762%,识别家蚕雌雄蛹的准确率为96.580%。特征融合图识别家蚕雌雄蛹的准确率比原始灰度图的提升了18.093%,改进VGG识别家蚕雌雄蛹的准确率比原始VGG的提升了2.257%。【结论】基于纹理特征和改进VGG的家蚕蛹雌雄识别方法能降低人工劳动时间,为实现家蚕蛹雌雄自动分拣提供基础。

蚕蛹多糖的碱液提取及免疫活性初步研究

以多糖含量为指标,以碱液浓度、提取温度、提取时间为实验因素和水平,采用正交实验设计,对蚕蛹多糖进行了碱液提取研究。结果表明,碱液提取蚕蛹多糖的最佳实验条件是碱液浓度0.02 mol.L-1、提取温度80℃、提取时间3 h。在此最佳实验条件下所得的蚕蛹多糖粗品中总糖含量为27.9%,氨基酸总量为35.85%,蛋白含量为38.1%。免疫活性实验显示,该多糖可以显著提高小鼠腹腔巨噬细胞吞噬指数和吞噬百分率、淋巴细胞转化率及血清溶血素,与对照组相比,差异具有显著性意义,表明蚕蛹碱提粗多糖可以明显增强小鼠的非特异性免疫、细胞免疫和体液免疫功能。

GC-MS分析蚕蛹油中脂肪酸的成分

目的：化学法提取的蚕蛹油经甲酯化后，分析其脂肪酸的成分。方法：用气相色谱质谱（ＧＣＭＳ）技术及图谱检索，对蚕蛹油中脂肪酸的成分进行鉴定和分析。结果：蚕蛹油中含９种脂肪酸，其中不饱和脂肪酸占７９．８８％，α亚麻酸的相对含量高达７１．４５％。结论：α亚麻酸具有降血脂、降血压和抗癌等生物功能，本研究为α亚麻酸的应用提供了重要的依据。

我国食品工业副产物资源化利用现状

我国是农业生产大国,农产品及食品工业的副产物数量巨大。为推进可持续化发展,这些副产物的资源化利用越来越受到人们的重视,并且很多副产物的深度开发与利用在研究与实践层面均取得了长足的进步,显示出良好的发展前景。以小麦、豆制品、油脂、蚕桑、石榴、葡萄等6大类食品加工业的副产物为例,系统阐述了我国小麦麸皮、小麦次粉、豆渣、豆粕、豆皮、油脚、脱臭馏出物、蚕蛹蛋白、蚕蛹油、石榴皮、石榴籽、葡萄皮、葡萄籽等食品工业废弃物的资源化利用现状。并以前期工作为基础,分别介绍了豆制品副产物及排放的黄浆水的资源化利用,石榴皮及石榴籽多酚提取物,石榴籽油及葡萄籽油的产品开发,桑蚕副产物制成的蚕蛹蛋白、活性肽、蚕蛹油、蚕蛹甲壳素的开发利用情况等。希望为我国食品工业副产物的深度开发与高值化利用提供借鉴和参考。

家蚕蛹蛾的营养组成评价分析

为明确不同性别家蚕蛹蛾资源的营养组成优势,通过比较其基本营养组成,分析蛋白的氨基酸和油脂的脂肪酸组成。结果发现:家蚕蛹蛾(干物质)的蛋白和脂肪含量超过80%,其中雄性脂肪含量较高,雌性蛋白和糖类含量较高,蚕蛹在羽化成蛾的变态过程中,水分、灰分、总糖含量显著下降。氨基酸组成分析发现,家蚕蛹蛾蛋白中必需氨基酸约占氨基酸总量的38%,其中雄性的赖氨酸和精氨酸含量明显高于雌性,氨基酸评分依次为雄蛾(100分)、雄蛹(94.8分)、雌蛹(80.9分)和雌蛾(73.1分);脂肪酸组成分析发现,家蚕蛹蛾不饱和脂肪酸的相对含量均在70%左右,其中雌性明显高于雄性。结果显示,家蚕蛹蛾是一种优质的昆虫油脂蛋白新资源,营养组成优势突出,形态性别差异明显。

蚕蛹蛋白过敏与综合利用的研究进展

蚕蛹蛋白是一种营养价值很高的天然动物蛋白,富含有18种氨基酸,其中八种人体必需氨基酸含量超过总氨基酸的40%,且比例均衡,具有非常高的利用价值,近年来被广泛利用于多个行业中。本文介绍了蚕蛹蛋白的种类和含量,进一步汇总了过敏原的相关信息,在此基础上总结了蚕蛹蛋白在食品、饲料、医药、纺织、生物等行业中的开发利用。

蚕蛹替代鱼粉对框鳞镜鲤幼鱼生长性能、体成分及健康状况的影响

为研究蚕蛹替代鱼粉对框鳞镜鲤幼鱼生长性能、体成分及健康状况的影响,试验配制含10%鱼粉的基础饲料(CP 36.18%,EE 5.18%)和蚕蛹替代50%鱼粉蛋白的蚕蛹饲料(CP 35.38%,EE 5.06%),饱食饲喂饲养于室内循环水养殖系统中的框鳞镜鲤幼鱼(12.10±0.89)g 61 d,日投喂3次。结果显示,蚕蛹组与鱼粉组的末体质量、增重率、特定增长率、饲料系数、成活率等生长性能指标,肥满度、内脏指数、脾脏指数、肝胰脏指数、肠指数等生物学性状指标,以及全鱼、肌肉、肝胰脏的一般营养成分均无显著性差异(P>0.05);蚕蛹组全鱼Lys水平和肌肉Tyr水平,肌肉C14∶0、C16∶1n-7、C18∶1n-7水平显著低于鱼粉组(P<0.05),C18∶3n-3、∑n-3PUFA水平显著高于鱼粉组(P<0.05),肝胰脏C18∶3n-3、C22∶5n-3水平显著高于鱼粉组(P<0.05),动脉粥样硬化指数(IA)和促凝血指数(IT)差异不显著(P>0.05);蚕蛹组和鱼粉组血清生化指标无显著性差异(P>0.05);蚕蛹组和鱼粉组的前肠和中肠肠道褶皱高度、褶皱密度、褶皱宽度、粘膜下层厚度没有明显差异(P>0.05),蚕蛹组前肠肌层显著较鱼粉组薄(P<0.05),中肠没有显著差异(P>0.05)。在本试验条件下,综合考虑其生长指标、体成分、血清生化指标及肠道组织结构形态,蚕蛹替代框鳞镜鲤日粮中50%鱼粉蛋白对机体氨基酸和脂肪酸组成有一定影响,但不影响框鳞镜鲤的健康状况,且有促进生长及饲料利用的趋势。