Host stadium specificity in the gregarine assemblage parasitizing Tenebrio molitor.

Reciprocal cross-stadia experimental infections were used to demonstrate stadium specificity within the gregarine assemblage parasitizing Tenebrio molitor, the yellow mealworm. Gregarina cuneata, Gregarina polymorpha, and Gregarina steini are characteristic parasites of larval T. molitor. Gregarina niphandrodes is a characteristic parasite of adult T. molitor. Experimental infections were produced in all homologous host-parasite combinations. No infection was produced in heterologous or cross-stadia combinations. This study introduces the concept of separate, distinct parasite niches corresponding to separate life cycle stages and established by known, predictable life cycle events within a single host species.     

Use of Tenebrio molitor (Coleoptera: Tenebrionidae) to recycle organic wastes and as feed for broiler chickens

Several dried waste materials from different origins were used as a substrate to grow Tenebrio molitor L. Nutrient/amino acid values differed depending on both larval size/weight and substrate. These larvae were experimentally used as a broiler feedstuff. Seven-day-old chicks of a commercially available strain with an average weight of 126 g were randomly distributed into nine six-broiler groups. Three levels of Tenebrio molitor larvae (0, 5, and 10% dry weight) were used in a 19% protein content sorghum-soybean meal basal diet, to evaluate feed intake, weight gain, and feed efficiency. Results after 15 d showed no significant differences among treatments. These data indicate that Tenebrio molitor has the potential to be used as protein source for raising broilers.     

An ultrastructural comparison of the attachment sites between Gregarina steini and Cryptosporidium muris

Early developmental stages of Gregarina steini Berndt, 1902 from the intestine of Tenebrio molitor larvae were studied by transmission electron microscopy. The formation and structure of the eugregarine attachment site were compared with comparable features found on the feeder organelle of Cryptosporidium muris Tyzzer, 1907, from the stomach of experimentally infected rodents. The similarity of the attachment strategy between both organisms was revealed. The membrane fusion site in G. steini, formed by the trophozoite plasma membrane, host cell plasma membrane and a membrane-like structure limiting the cortical zone of the epimerite, resembles the Y-shaped membrane junction between the host cell plasma membrane, the trophozoite plasma membrane and membrane surrounding the anterior vacuole in C. muris. The anterior vacuole of C. muris appears to be the precursor of the feeder organelle and its structure is very similar to the epimeritic bud and the cortical zone of G. steini trophozoites. In both investigated organisms, the apical complex disappears early during cell invasion. The possibility of the epicellular location of Cryptosporidium on the surface of host cells is discussed.     

温度对黄粉虫幼虫生长发育及体液免疫防御能力的影响

【目的】探究饲养温度对黄粉虫Tenebrio molitor幼虫生长发育和体液免疫防御的影响。【方法】测定了不同温度（18, 22, 26和30℃）下饲养的黄粉虫幼虫的发育历期、蛹重、化蛹率;采用抑制区分析法测定了不同温度下饲养的免疫（用生理盐水将大肠杆菌Escherichia coli配制成1×104个菌体/μL悬浮液,用微量注射器将其注入虫体腹部的背面,每头幼虫注射1 μL）和非免疫(注射生理盐水)黄粉虫幼虫血淋巴的抑菌和溶菌酶活性,通过分光光度法测定了其酚氧化酶活性。【结果】结果显示,黄粉虫幼虫发育历期随饲养温度的上升而明显缩短（P<0.0001）,而不同温度下蛹重（P=0.067）与化蛹率（P=0.869）差异不显著。免疫组黄粉虫幼虫血淋巴的抑菌、酚氧化酶和溶菌酶活性随饲养温度上升而降低：抑菌和酚氧化酶活性随温度变化差异极显著（P<0.0001）,溶菌酶活性差异显著（P=0.013）。【结论】本研究结果表明,温度对黄粉虫的生长发育和免疫防御具有较大的影响,低温下黄粉虫幼虫的发育历期延长,但其体液免疫防御能力明显增强。     

Comparative biochemical study of brain phospholipids in insects

Studies have been made on the content of total phospholipids (PL) and their separate families in the brain of larvae and imago of the insect species Blaberus giganteus, Periplaneta americana, Tenebrio molitor and Barathra brassicae. It was shown that during larval-pupal-imaginal transformation in T. molitor and B. brassicae the content of total PL increases by 17 and 14% respectively, whereas in B. giganteus and P. americana PL concentration undergoes only insignificant changes. With respect to total PL content the species investigated form the following sequence: B. brassicae greater than T. molitor greater than P. americana greater than B. giganteus. In larval and imago forms of these insects phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, sphingomyelin and polyglycerophosphatide were found in the brain. Besides, in the brain of B. giganteus and P. americana lysophosphatidylethanolamine and in the brain of T. molitor -- phosphatidic acid are present. Qualitative PL composition in larval and imago forms is identical, while quantitative ratio of separate fractions differs insignificantly. In all the species investigated, phosphatidylethanolamine and phosphatidylcholine are the most abundant phospholipids.     

The precocious commitment of wing Anlagen in Tenebrio molitor revealed by the addition of 20-hydroxyecdysone

An injection of 20-hydroxyecdysone (10 mug per animal) 6-13 days after the moult of the last larval instar of Tenebrio molitor induces the development of prothetelic larvae and larval-pupal intermediates. The state of larval-pupal switchover, or commitment, is only disclosed at the time of injection of the moulting hormone. Prothetelic A and B larvae, with small and medium sized wing Anlagen, undergo another larval or pupal instar. Prothetelic C larvae with bigger Anlagen are unable to moult, but the adult programme is expressed. Ecdysed larval-pupal intermediates give more or less perfect adults, while unecdysed mealworms, imprisoned in their larval cuticle, also expressed the adult programme. The commitment of Tenebrio is not a global switchover because a significant asynchronisation is noted between the development of organs considered. Animal crowding induces a delay in the appearance of wing Anlagen.     

Individual and population effects of eugregarine, Gregarina niphandrodes (Eugregarinida: Gregarinidae), on Tenebrio molitor (Coleoptera: Tenebrionidae)

Gregarines are single-celled parasites in the phylum Apicomplexa that infect invertebrates. They are highly abundant on three levels: among a large diversity of invertebrates, in the proportion of population of organisms they infect, and within individually infected organisms. Because of their remarkable prevalence, we hypothesize that they play an important role in support of their hosts. However, studies done to date on the impact of gregarines on their host are conflicting. Therefore, we studied the impact of gregarines on their host using a model Gregarina niphandrodes infection in Tenebrio molitor. The impact of infection was measured by comparing beetles with no or low infection to those with artificially induced high infection. The numbers of individuals in each of the three easily visible developmental stages of the T. molitor (larva, pupa, and adult) were censused weekly. From these observations, fertilities and probabilities of survival with transition between stages were estimated. These estimated vital rates were used to construct a stage-classified projection matrix model. We also measured the longevity of individual beetles with low and high infection that were grown in isolation. The results indicate that there is no significant difference in the population dynamics of beetles with low and high infection. However, the longevity was significantly different between beetles with low infection than the deliberately highly infected group.     

Constant topological organization of the coleopteran metamorphosing nervous system: analysis of persistent elements in the nervous system of Tenebrio molitor

Evidence is provided that the topological organization of the larval neuropil is preserved during metamorphosis of Tenebrio molitor. Constancies in neuronal organization were due in part to persistence of individual neurons whose morphologies were individually followed. It appears that the phenomenologically static situation of the metamorphosing neuropil is achieved by stabilization and regulation due to cellular interactions. Comparisons are made with features of hemimetabolan postembryogenesis.     

Localization of post-proline cleaving peptidases in Tenebrio molitor larval midgut

Two soluble post-proline cleaving peptidase activities, PPCP1 and PPCP2, were demonstrated in Tenebrio molitor larval midgut with the substrate benzyloxycarbonyl-L-alanyl-L-proline p-nitroanilide. Both activities were serine peptidases. PPCP1 was active in acidic buffers, with maximum activity at pH 5.3, and was located mainly in the more acidic anterior midgut lumen. The dynamics of PPCP1 activity and the total activity of soluble digestive peptidases in the course of food digestion were similar, suggesting that the enzyme participates in protein digestion. PPCP2 is a nondigestive soluble tissue enzyme evenly distributed along the midgut. An increase in the activity of PPCP2 was observed in buffers of pH 5.6-8.6 and was maximal at pH 7.4. The sensitivity of PPCP2 to inhibitors and the effect of pH are similar to prolyl oligopeptidases with a cysteine residue near the substrate binding site.     

泰山虫草对黄粉虫的侵染特性研究

报道了用泰山虫草分生孢子悬浮液接种活体黄粉虫,提取被侵染的黄粉虫体液,并测定不同侵染时间虫体液中酶活性的动态变化。结果表明:泰山虫草分生孢子能成功的侵染黄粉虫并长出子座;接种第2~4 d后即可导致黄粉虫体内活性发生不同程度的变化,说明黄粉虫具有独特的防御病原微生物的侵染机制。     

饲喂黄顶菊对黄粉虫生长发育及繁殖的影响

【背景】黄顶菊是近年来人侵我国的一种有害杂草,其竞争力强,给自然界造成了严重危害,同时也产生了大量的生物质资源。作为一种杂食性昆虫,黄粉虫易于饲养,可以用于多种生物质资源的转化处理。【方法】本研究分别以黄顶菊、小白菜和西瓜皮作为青料,与麦麸等配制成人工饲料后,对黄粉虫进行定期饲喂,研究其对黄粉虫幼虫、成虫生长发育及繁殖的影响。【结果】黄粉虫幼虫饲料中添加一定比例的黄顶菊与添加小白菜相比,虫体生物量增长率较低,饲料利用率和死亡率较高;成虫饲料中添加黄顶菊与添加西瓜皮相比,黄粉虫平均寿命延长,单雌产卵量减少,且差异均显著,体长略有减小。【结论与意义】本研究对利用黄粉虫转化处理黄顶菊的效果做出了初步评价,即黄粉虫能够取食黄顶菊,但转化处理效果不太理想。     

The role of amino acid residues in the active site of a midgut microvillar aminopeptidase from the beetle Tenebrio molitor

Aminopeptidases are major enzymes in the midgut microvillar membranes of most insects and are targets of insecticidal Bacillus thuringiensis crystal delta-endotoxins. Sequence analysis and substrate specificity studies showed that these enzymes resemble mammalian aminopeptidase N, although information on the organization of their active site is lacking. The effect of pH at different temperatures on the kinetic parameters of Tenebrio molitor (Coleoptera) larval aminopeptidase showed that enzyme catalysis depend on a deprotonated (pK 7.6; DeltaH degrees (ion), 7.6 kJ/mol) and a protonated (pK 8.2; DeltaH degrees (ion), 16.8 kJ/mol) group. 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide and diethylpyrocarbonate inactivate the enzyme by modifying a pK 5.8 carboxylate and a imidazole group, respectively, with a reaction order around 1. Tetranitromethane changes the K(m) of the enzyme without affecting its V(max) by modifying a phenol group. The presence of a competitive inhibitor decrease the inactivation reaction rates in all these cases. EDTA inactivation of the aminopeptidase is affected by pH and temperature suggesting the involvement in metal binding of at least one deprotonated imidazole group (pK 5.8, DeltaH degrees (ion), 20 kJ/mol). The data support the hypothesis that T. molitor aminopeptidase catalysis depends on a catalytic metal and on a carboxylate and a protonated imidazole group, whereas substrate binding relies in one phenol and one carboxylate groups. The insect aminopeptidase shares common features with mammalian aminopeptidase N, although differing in details of substrate binding and in residues directly involved in catalysis.     

Alpha-galactosidases from the larval midgut of Tenebrio molitor (Coleoptera) and Spodoptera frugiperda (Lepidoptera)

There are three midgut alpha-galactosidases (TG1, TG2, TG3) from Tenebrio molitor larvae that are partially resolved by ion-exchange chromatography. The enzymes have approximately the same pH optimum (5.0), pl value (4.6) and Mr value (46000-49000) as determined by gel filtration or native electrophoresis run in polyacrylamide gels with different concentrations. Substrate specificities and functions were proposed for the major T. molitor midgut alpha-galactosidases (TG2 and TG3) based on chromatographic, carbodiimide inactivation, Tris inhibition, and on substrate competition data. Thus, TG2 would hydrolyse alpha-1,6-galactosaccharides, exemplified by raffinose, whereas TG3 would act on melibiose and apparently also on digalactosyldiglyceride, the most important compound in the thylacoid membranes of chloroplasts. Most galactoside digestion should occur in the lumen of the first two thirds of T. molitor larval midguts, since alpha-galactosidase activity predominates there. Spodoptera frugiperda larvae have three midgut alpha-galactosidases (SG1, SG2, SG3) partially resolved by ion-exchange chromatography. The enzymes have similar pH optimum (5.8), pl value (7.2) and Mr value (46000-52000), and at least the major alpha-galactosidase must have an active carboxyl group in the active site. Based on data similar to those described for T. molitor, SG1 and SG3 should hydrolyse melibiose and SG3 should digest raffinose and, perhaps, also digalactosyldiglyceride. The midgut distribution of alpha-galactosidase activity supports the proposal that alpha-galactosidase digestion occurs at the surface of anterior midgut cells in Spodoptera frugiperda larvae.     

cDNA cloning, expression, and characterization of an arylphorin-like hexameric storage protein, AgeHex2, from the mulberry longicorn beetle, Apriona germari

An arylphorin-like hexameric storage protein, AgeHex2, cDNA was cloned from the mulberry longicorn beetle, Apriona germari (Coleoptera, Cerambycidae), larval cDNA library. The complete cDNA sequence of AgeHex2 is comprised of 2,088 bp encoding 696 amino acid residues. The AgeHex2 had four potential N-glycosylation sites. The AgeHex2 contained the highly conserved two larval storage protein signature motifs. The deduced protein sequence of AgeHex2 showed high homology with A. germari hexamerin1 (51% amino acid identity), Tenebrio molitor hexamerin2 (49% amino acid identity), T. molitor early-staged encapsulation inducing protein (43% amino acid identity), and Leptinotarsa decemlineata diapause protein1 (43% amino acid identity). Phylogenetic analysis further confirmed the AgeHex2 is more closely related to coleopteran hexamerins than to the other insect storage proteins. Northern blot analysis confirmed that the AgeHex2 showed fat body-specific expression. The cDNA encoding AgeHex2 was expressed as a 75-kDa protein in the baculovirus-infected insect cells. Furthermore, N-glycosylation of the recombinant AgeHex2 was revealed by tunicamycin to the recombinant virus-infected Sf9 cells, demonstrating that the AgeHex2 is N-glycosylated. Western blot analysis using the polyclonal antiserum against recombinant AgeHex2 indicated that the AgeHex2 corresponds to a 75-kDa storage protein present in the A. germari larval hemolymph.     

黄粉虫成虫繁殖力及影响幼虫发育的因素

对单配或一雄多雌或经历不同交配历期的雌黄粉虫 Ｔｅｎｅｂｒｉｏ ｍｏｌｉｔｏｒ Ｌ．产卵量进行了研究,以期了解两性的繁殖力,为雌虫多产高质量卵提供科学依据。在幼虫期添加少量鲜马铃薯可极大地促进其生长。适当的高密度和不筛粪有利于低龄幼虫生长,４０日龄以后应定期筛粪并添加新饲料。幼虫的最适密度取决于其拥有的饲料量。幼虫生态生长效率高达３１．８％。     

Beetle-to-beetle transmission and dispersal of Hymenolepis diminuta (Cestoda) eggs via the feces of Tenebrio molitor

When grain beetles (Tenebrio molitor) were fed eggs of Hymenolepis diminuta, many of the eggs passed intact through the beetles' intestines, and eggs were present in the beetles' feces for at least 48 hr after feeding. When uninfected T. molitor were fed beetle feces containing H. diminuta eggs, they became infected. Tenebrio molitor were fed on H. diminuta eggs and then placed in fresh bran for 48 hr. When uninfected T. molitor were placed in this bran, they became infected. Thus, feces from beetles that have ingested H. diminuta eggs serve as a source of eggs for other beetles, as well as a mechanism of egg dispersal.