Comparison of Nucleic Acid Content among Non-diapause Pupae, Diapause Pupae and Eclosion-adult from Diapause Pupae of Papilio memnon

[Objective] The aim of this study was to provide basis for deeply understanding the diapause mechanism of Papilio memnon L. [Method] RNA and DNA content of non-diapause pupae, diapause pupae and eclosion-adult from diapause pupae at different development stages were detected by the colorimetry. [Result] RNA content of non-diapause pupae was 4.614 0-7.946 3 μg/mg, while diapause pupae was 4.326 0-5.885 3 μg/mg and eclosion-adult from diapause pupae was 20.779 3 μg/mg at initial stage. DNA content of non-diapause pupae was 0.448 7-0.535 0 μg/mg, while diapause pupae was 0.452 0-0.828 3 μg/mg and eclosion-adult from diapause pupae was 1.727 0 μg/mg at initial stage. [Conclusion] The nucleic acid content and change is related to the development stage.

Comparison of Protein Content among Non-diapause Pupae,Diapause Pupae and Eclosion-adult from Diapause Pupae of Papilio memnon

[ Objective] The experiment aimed to explore the diapause mechanism of Papilio memnon L. further. [ Method] The colorimetry method was used to detect protein content of non-diapause pupae, diapause pupae and eclosion-adult from diapause pupae at different developmental stages. [Result] It was showed that the protein content of non-diapause pupae was from 111.768 to 115.030 7 mg/g and the change of protein content during whole pupal stage, while diapause pupae were from 81.218 to 131.623 7 mg/g and the change of protein content was not big from last ten-day period of October to last ten-day pariod of November, however, the content increased from the last ten-day period of November. The protein content of emerging adult was 296.969 3 mg/g. [Conclusion] The change of protein content was related to Papilio memnon at different development stage.

Developmental Threshold Temperature and Effective Accumulative Tem- perature of Pupae and Eggs of Holcocerus hippophaecolus

In order to understand the occurrence and the developmental regularity of seabuckthorn carpenterworm (Holcocerus hippophaecolus) and predict its population density, the developmental threshold temperature (C) and effective accumulative tem- perature (K) of the carpenterworm pupae and eggs were analyzed under the conditions of constant and variable temperatures. The results show that the values of C and K of the carpenterworm pupae are (12.1 ± 0.2) °C and (295.2 ± 4.1) day-degrees at constant temperatures, and (15.5 ± 0.4) °C and (202.4 ± 13.1) day-degrees at variable temperatures. However, the values of C and K of the eggs at variable temperatures are (16.7 ± 0.8) °C and (101.5 ± 12.6) day-degrees. The differences of developmental threshold and effective accumulative temperature under the conditions of constant and variable temperatures of the carpenterworm pupae accord with the developmental regularity of most insects in nature. By comparing five different constant temperatures, the conclusion is that the optimum developmental temperature of the pupae is 21 °C when both the pupation of the mature larvae and the eclosion of the pupae are very accordant. Moreover, the percentage of eclosion is over 90%. The average developmental durations of the carpenter- worm pupae and eggs are 31 and 16 d at variable temperatures.

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.

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.

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.

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.

Ant Pupae from Myanmar Amber Reveal the Evolutionary Process of Cocoon-Spinning Behavior in Formicidae

Many extant species spin cocoons before their pupal stage,which can protect them from predators,pathogens,and fluctuant environmental factors in nests.However,owing to the scarcity of ant cocoon fossils,little is known about the origin and evolution of this behavior in Formicidae.Here,we describe two ant pupae specimens from Myanmar amber.The presence of cocoons in these Mesozoic fossils suggests the ancestral origin of spinning cocoon behavior;based on our phylogenetic analysis and ancestral state reconstruction,this behavior was secondarily lost at least six times in Formicidae,which could be ascribed to the transition of nesting and feeding habits,as well as the development of eusociality.Moreover,the first specimen preserved worker ants with empty cocoons simultaneously,indicating garbage removal or brood care behavior,which improves our understanding of their eusociality in the Cretaceous.

INHIBITION EFFECTS OF ANTIBACTERIAL PEPTIDE D FROM THE CHINESE OAK SILKWORM(Antheraea pernyi) PUPAE

Some active antibacterial substances, such as antibacterial peptide lysozymes and agglutinin were induced and developed from the haemolymph of the Chinese oak silkworm(Antheraea pernyi) pupae injected with Escherichia coli or treated with ultrasonic waves. The antibacterial peptide from A. pernyi was isolated, purified, and its amino acid sequence was also determined. The inducible antibacterial substances have marked

DETERMINATION OF JUVENILE HORMONE TITER IN THE HEMOLYMPH OF PUPAE OF INDIAN SILKWORM, Philosamia cynthia ricini

Juvenile hormone plays an important role in controlling the processes of metamorphosis and reproduction of most insects. There has been a growing amount of research works on the determination of JH titers at various physiological levels in a variety of insect species, yet our knowledge concerning JH titer lags behind that of ecdysteroid titer. The reasons are: (ⅰ) JH titers in most insects seem to be generally quite low, and (ⅱ) it is difficult to separate JH from the lipids that are present in rather large quantities and have similarities in chemical and physical properties.

Spatial distribution of Holcocerus hippophaecolus (Lepidoptera: Cossidae) pupae in a seabuckthorn (Hippophae rhamnoides) stand

The seabuckthorn carpenter moth,Holcocerus hippophaecolus,which has a generation time of four years,is recently becoming one of the major pests of the seabuckthorn(Hippophae rhamnoides)in Inner Mongolia,Liaoning,Shanxi,Ningxia and Shaanxi of China(Hua et al.,1990).The larvae of the H.hippophaecolus mainly damage the stems and roots of the seabuckthorn,and the mature larvae pupate in the soil.The spatial distribution of the pupae was analyzed by using biostatistics and geostatistics in order to effectively control the insect and further study the spatial distribution of the population.Results show that most of the pupae(90%)had an eclosion time span from early June to the end of July.The sex ratio of the pupae was nearly 1:1 in the woodland samples.In addition,24.3%of the 971 trees investigated had pupae and it ranged from 0 to 4 per tree within a distance of 1.3 m from the base of the stem.90%of the pupae were aggregated within a distance of 1 m from the base of the stem.The pupae show intense spatial aggregation in the sampled woodland which had an 11.1 m spatial dependence and a 90.7%intensity in the local spatial continuity.Moreover,the population presented an intensive spotted distribution and many aggregated spots were found in the woodlands.As for the relationship between grid size and variogram of the pupae,the variations in the range,the intensity of local spatial continuity and the sill were all very low or non-existent when the grid size was 5 m,6 m or 7 m.Whereas,the value of the decisive coefficient was the biggest when the grid size was 5 m making it the ideal grid size.

Gastrointestinal digestion fates of lipids based on pH-stat and Caco-2 cells models:Superior bioavailability of triglyceride monomer derived from silkworm pupae oil

Silkworm pupae oil(SPO)was regarded as a good alternative to alpha-linolenic acid supplements,which contain a variety of triacylglyceride with high nutritional value.To investigate the connections between triacylglyceride composition and lipid gastrointestinal digestive absorption profiles,isolation and bioavailability of triacylglyceride monomers from silkworm pupae oil was analyzed.In this study,two triacylglyceride monomers rich inα-linolenic acid were separated by preparative liquid chromatography,namely LnLnLn(1,2,3-α-linolenic acid triacylglyceride)and LnLnL(1,2-α-linolenic acid,3-linoleic acid triacylglyceride).The in vitro digestion in the pH-stat model results showed the digestive rate of LnLnLn was 0.4101±0.0089 s^(-1),which was significantly higher than that of SPO(0.2494±0.0108 s^(-1))(p<0.05).The in vitro absorption in the Caco-2 cells model results showed the absorptivity of LnLnLn and LnLnL were 6.40±0.19%and 6.06±0.54%,which were significantly higher than SPO(5.43±0.51%)(p<0.05).In addition,the mRNA expression levels of lipid metabolism-related proteins provide evidence for the absorption superiority of LnLnLn and LnLnL.The present work provides novel structural lipids for rapid and efficient supplementation ofα-linolenic acid.

Effective accumulated temperature and developmental threshold temperature for Semanotus bifasciatus(Motschulsky) in Beijing

In order to understand the occurrence and development of Semanotus bifasciatus （Motschulsky）, a borer insect of Platycladus orientalis in parts of northern and eastern China and to improve its prediction, the developmental threshold temperature （C1） and effective accumulated temperature （K） of S. bifasciatus were determined under conditions of constant and variable temperatures. The results show that under constant temperatures the value of C1 has a range of 8.90±1.77℃and the value of K lies between 95.19±13.14 degree-day for eggs; the values for larvae are C1= 13.26±3.06℃ and the number of degree-day is K 2,885.07±187.87 degree-day; for pupae the ranges are C1 = 8.93±2.49℃ and K= 131.20±25.63 degree-day. Under the condition of ambient temperatures, the values for eggs are C1= 17.33±1.24℃ and K= 70.79±8.99 degree-day. It is suggested that S. bifasciatus adults over-winter in a state of dormancy, not at a diapause. Warm winter may accelerate the life cycle ofS. bifasciatus. Equations can be used to predict an early occurrence ofS. bifasciatus.

Effects of Sublethal Doses of Chlorfluazuron on Pupal-testis in the Common Cutworm, Spodoptera fitura （F,） （Lepidoptera： Noctuidae）

The effects of sublethal doses （LD10：1.00 ng larval-1; LD30： 3.75 ng larva-1） of chlorfluazuron on number and size of developmental spermatocytes in pupal-testis during spermatogenesis of the common cutworm, Spodoptera litura （F.） are described. Chlorfluazuron was applied topically to newly-ecdysed fifth-instar larvae of S. litura under laboratory conditions. During first day of pupation, spermatids were the highest （3,430 ± 7）, however, elongated spermatocytes with mature sperm were the lowest （292 ± 16） in number in pupal-testis compared with other developing spermatocytes. During last day of pupation, elongated spermatocytes with mature sperm were the highest （3,581 ± 3）, however, spermatogonia were the lowest （86± 4） in number in pupal-testis compared with other developing spermatocytes. The number of spermatogonia, primary and secondary spermatocytes, spermatids and elongated cysts with mature sperm were increased during pupal-developmental days in the controls. However, their numbers were significantly reduced in LD10- and even more significantly reduced in LD30-treated pupal-testis compared with controls. Moreover, eupyrene sperm bundles were not present in pupal-testis. During pupal-developmental days, spermatids were the smallest （4.1 ± 0.9 dm in μm） and secondary spermatocytes were the largest （31.0 ± 1.8 dm in μm） in size compared with other developing spermatocytes. The sizes of the same developing spermatocytes were significantly reduced in LD10-treated and more significantly reduced in LD30-treated pupal-testis of S. litura compared with controls.

Study on Pupa Changes and Emergence Behaviors of Dacne picta Crotch

[ Objective ] The research aimed to study the pupa changes and emergence behaviors of Dacne picta Crotch. [ Method ] The D. p/cta reared in the labo- ratory were observed and recorded. [ Result ] Before the emergence, the color of compound eyes, maxillae, tarsi and wing pads d pupae was gradually deepened. The 14^th- 16^th d after pupation were the peak time for emergence, and the ratio of female to male was close to 1： 0.77. During the emergence, the puparium of D. picta dehisced from pronotum, and the hind wing had extension phenomenon before contraction and folding. [ Conclusion] The research confirmed the pupa changes and the emergence behaviors of D. picta.

Effect of the Variety of Fig Tree on Some Biological Parameters of Ceratitis capitata Wied, 1824 （Diptera： Trypetidae） in Some Orchards in the Kabylie

The Mediterranean fruit fly, Ceratitis capitata Wied. （1824） is one of the most important pest that can infest orchards if its spread is not controlled. Among the preventive measures recommended in the fight against this pest, we are interested in looking for varieties that are less favorable to the development of this pest among different varieties of fig trees in our region. The biological parameters of the fruit fly which were followed in this study are duration of pupation, rate of emergence, sex ratio and longevity of males and females. The results show that these parameters vary depending on the variety of fruit.

Morphological traits for sex identification of the oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)

The oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)is a harmful pest which causes heavy loss in agricultural crops.Sex identification of M.separata is very important for understanding the biology and management of this pest.Development of fast and convenient methods to identify M.separata female and male is the necessary prerequisite for con ducting gender-related studies.In this study,new methods to differentiate sex of M.separata at pupal and adult stages were investigated.Our results showed that posterior abdominal segments containing genital organs which are morphological traits to differentiate female and male.Genital openings are important characters for the female and male pupae differentiation.For female pupa,it locates on the ventral side of eighth and ninth abdominal segments and appears as a visible longitudinal suture;for male pupa,it locates on the ventral side of the ninth abdominal segment and appears as a short slit surrounded by a pair of visible semicircular lumps.Bristle numbers are another indicator for sex differentiation of adults.Each female has three-bristled frenulum and each male has one bristled-frenulum near the base of hind wing of each side of moth body.We confirmed the two sex differentiation methods manually by dissecting genitalia of adults and found their genger was as same as what we examined the sex of pupae and adults using the morphological methods.

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.

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.

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.