Antioxidant system in the darkling beetle (Tenebrio molitor) in ontogenesis

The level of antioxidant protection and lipid peroxidation (LP) intensity in the darkling beetle Tenebrio molitor on different developmental stages were assessed. Each stage was shown to be characterized by its own peculiarities of prooxidant-antioxidant balance. Thus, maximal intensity of oxidative processes estimated by LP intermediate product (diene conjugates and ketodiens) accumulation is attributable to pupae, and minimal intensity--to the 3rd-5th instar larvae. Superoxide dismutase activity increases gradually during the life cycle. A decline in catalase (CAT) and glutathione reductase (GR) activities occurred on the stage of pupae. CAT activity in imago was equal to the larva values, and GR activity in imago even exceeded the larva values. At the same time GR activity in T. molitor was detected only at 37 degrees C under our experimental conditions. No statistically significant changes in glutathione reduced content were observed in the insects during the life cycle.     

Effect of Feeding on Three Eucalyptus Species on the Development of Brontocoris tabidus (Het.: Pentatomidae) Fed with Tenebrio molitor (Col.: Tenebrionidae)

Brontocoris tabidus (Heteroptera: Pentatomidae) is an important predator of defoliating Lepidoptera in Brazil, which has been mass produced and released against defoliating caterpillars in Eucalyptus plantations. We aimed to improve the mass production of this predator by providing eucalypt seedlings, in addition to an alternative prey, for its development. The effect of three Eucalyptus species on the development of B. tabidus fed with Tenebrio molitor L. (Coleoptera: Tenebrionidae) pupae, was studied at a temperature of 25.0 ±2.5 o C; a relative humidity of 70.0 ±10.0% and a photophase of 12:12 hours (light:dark). Pupae of T. molitor were provided in each of the following feeding treatments: treatment 1, Eucalyptus urophylla seedlings; treatment 2, Eucalyptus camaldulensis seedlings; treatment 3, Eucalyptus grandis seedlings; and treatment 4, no seedling (only pupae of T. molitor ). Duration of the nymphal phase of the predator was similar in all treatments with plant and prey. It was observed that adults developed from nymphs fed on T. molitor only, had deformed wings and a small abdomen. B. tabidus fed during their nymphal and adult stages with prey together with Eucalyptus seedlings, had improved longevity, higher egg viability and a larger number of eggs and nymphs, than those insects fed only with prey. Therefore, it seems advisable to rear B. tabidus with prey and seedlings of Eucalyptus species, to improve the mass production of this predator for biological control of defoliating caterpillars of eucalypt plantations.     

室内条件下莱氏猛叩甲对松墨天牛和黄粉虫的捕食能力

【目的】本研究旨在明确莱氏猛叩甲Tetrigus lewisi老熟幼虫分别对黄粉虫Tenebrio molitor和松墨天牛Monochamus alternatus末龄幼虫和蛹的捕食量的差异,评估莱氏猛叩甲的捕食能力,为松墨天牛M.alteratus的有效治理找到潜在的高效生物防治因子。【方法】于2014年11月在浙江富阳昌东村马鞍山砍下松墨天牛虫害木,通过林间野外调查,明确松墨天牛和莱氏猛叩甲的发生情况;将野外采集到的莱氏猛叩甲幼虫带回室内,分别饲喂黄粉虫和松墨天牛的末龄幼虫和蛹,每天记录捕食量,确定莱氏猛叩甲的捕食能力。【结果】野外调查中,我们在松墨天牛幼虫蛀食的坑道中发现了莱氏猛叩甲幼虫和松墨天牛幼虫被取食后的残体;在采集到1 094头松墨天牛幼虫的危害木内,共采得莱氏猛叩甲末龄幼虫36头,约为松墨天牛幼虫数量的3%。室内捕食实验表明,5 d内莱氏猛叩甲捕食黄粉虫幼虫总量为3.2头,平均每天捕食约0.6头,而捕食松墨天牛幼虫总量为8.0头,平均每天捕食约1.6头,莱氏猛叩甲对松墨天牛的总捕食量显著高于对黄粉虫幼虫的总捕食量(P0.0001);4 d内莱氏猛叩甲捕食黄粉虫蛹总量为5.6头,平均每天捕食约1.4头,而捕食松墨天牛蛹总量为7.8头,平均每天捕食约2.0头,莱氏猛叩甲对松墨天牛蛹的捕食量显著高于黄粉虫蛹(P=0.028)。回归分析还表明,莱氏猛叩甲幼虫的捕食量不受叩甲个体大小的影响。【结论】莱氏猛叩甲普遍分布在松墨天牛发生区,同时发现松墨天牛被取食后的残体,表明莱氏猛叩甲是松墨天牛的捕食性天敌。莱氏猛叩甲均能捕食黄粉虫和松墨天牛的幼虫和蛹;相比于捕食黄粉虫,莱氏猛叩甲对松墨天牛的幼虫和蛹具有更好的捕食效率,具有很好的生物防治前景,是未来松墨天牛治理潜在高效生物防治因子。     

Development of the Predatory Stinkbug Brontocoris tabidus Signoret Heteroptera: Pentatomidae on Different Proportions of an Artificial Diet and Pupae of Tenebrio molitor L. Coleoptera: Tenebrionidae

The development of the Neotropical predatory pentatomid Brontocoris tabidus on an artificial diet based on beef meat and liver was evaluated. The predator showed significantly longer nymphal development and lower adult weights on this diet than when reared on pupae of the mealworm Tenebrio molitor. The survival of nymphs fed exclusively on the artificial diet was somewhat lower compared with feeding on T. molitor pupae. When B. tabidus was bred on this artificial diet during part of its nymphal period i.e. during the second; second and third; and second, third and fourth instars , and was subsequently returned to T. molitor pupae, the predator nymphs completed the nymphal stage with a developmental rate similar to that of nymphs fed on live prey throughout. The adults attained after switching from the artificial diet to live prey from the third and fourth instar onwards had similar weights to those in the control. Considering the relatively good results obtained with B. tabidus and other members of the pentatomid subfamily Asopinae, this meat-based diet may be a valuable alternative for use in the mass production of predatory pentatomids.     

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.     

Tenebrio beetle pupae show a conditioned behavioural response to pulse rotations of a geomagnetic field

Abstract.  Pupae of holometabolous insects are not motionless and insensitive developmental stages. Their behavioural display is restricted to rotations or contractions of the abdomen in a range of movements from the clearly visible to the microscopic of the order of tenths of microns. Pupae react spontaneously and surprisingly sensitively to mechanical, light or sound stimuli. In the present study, reactions of yellow mealworm beetle ( Tenebrio molitor , L.) pupae to a geomagnetic field rotation are examined. By means of a micromechanical recording technique, peaks of abdominal contractions are monitored before and after magnetic treatment and show that spontaneous behavioural reaction to a magnetic pulse is insignificant. Nevertheless, using negative reinforcement training, a conditioned magneto-sensitive reaction is elicited. These surprising sensory and behavioural capacities of an insect pupa are discussed.     

Nymphal Development and Reproduction of Podisus nigrispinus (Heteroptera: Pentatomidae) Fed With Combinations of Tenebrio molitor (Coleoptera: Tenebrionidae) Pupae and Musca domestica (Diptera: Muscidae) Larvae

The development and reproduction of Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) fed on Tenebrio molitor L. (Coleoptera: Tenebrionidae) and Musca domestica L. (Diptera: Muscidae) were studied in four treatments. P. nigrispinus was permitted to feed on: (1) T. molitor pupae (T 1 ); (2) Musca domestica larvae (T 2 ); (3) both prey supplied simultaneously (T 3 ); (4) both prey supplied on alternate days (T 4 ). Duration of the nymphal period of P. nigrispinus was similar in all diets studied with nymphal viability of approximately 75%. Heavier females were obtained in T 1 and T 4 , but no correlation between this factor and the reproductive rate of the predator was found. Therefore, the use of body weight as a parameter to evaluate rearing quality should be approached with caution. However, females of this predator showed higher egg and nymph production when they received both prey. For this reason P. nigrispinus should be reared with T. molitor and M. domestica simultaneously or on alternate days.     

Effect of weak combined magnetic fields on the metamorphosis of the meal-worm beetle Tenebrio molitor

The effects of weak combined magnetic fields adjusted to the parametric resonance for Ca2+ and K+ and extremely weak alternating magnetic field on the metamorphosis of the meal-worm beetle Tenebrio molitor have been studied. It was shown that the exposure of pupas of insects to all above-indicated types of fields stimulates the metamorphosis. However, after the exposure to weak combined magnetic fields adjusted to the parametric resonance for Ca2+ and K+, the number of insects with anomalies increases, which is not observed by the action of the weak alternating magnetic field.     

藁本提取物杀虫活性初步研究

分别采用点滴法、三角瓶熏蒸法和小叶碟添加法测定了藁本（Ligusticum sinense Oliv．）提取物对粘虫（Mythimna separata Walker）和小菜蛾（Plutella xylostella L．）幼虫的触杀、熏蒸和拒食活性,同时也测定了对黄粉虫（Tenebrio molitor L．）蛹的生长发育有抑制作用,并对其致毒症状、作用方式进行了初步探讨。结果表明,不同溶剂提取物中,丙酮冷浸提取物对粘虫毒杀活性较高;其丙酮提取物对粘虫和小菜蛾3龄幼虫均具有较高触杀及熏蒸活性,24h触杀LD50分别为0．062 3mg干样／头和0．150 2mg干样／头;12h熏蒸LC530分别为24．3422 mg干样／L和10．601 5mg干样／L;对黄粉虫（Tenebrio molitor L）蛹的生长发育有抑制作用,ID50为6．5157mg干样／头;粘虫中毒后先后表现出兴奋、痉挛、麻痹等症状,最后死亡;对黄粉虫蛹生长发育的抑制作用类似于保幼激素活性。     

黄粉虫不同虫态肠道细菌分离及鉴定

目的研究黄粉虫肠道细菌的类群与其取食的关系,为进一步开发利用提供理论依据。方法从人工饲养的黄粉虫幼虫、蛹和成虫肠道环境中分离纯化获得9个细菌菌株,对其菌体形态、染色反应、培养性状、生理生化反应进行系统研究。结果研究结果表明,上述9个细菌菌株分别属于放线杆菌属（Actunobacillus）、丙酸杆菌属（Propionibacterium）、柠檬酸杆菌属（Citrobacter）、沙雷菌属（Serratia）、芽胞杆菌属（Bacillus）、皮杆菌属（Dermabacter）、短状杆菌属（Brachybacterium）、棍状杆菌属（Clavibacter）和微小杆菌属（Exiguobcacterium）。结论通过对黄粉虫不同虫态即幼虫、蛹和成虫肠道细菌分离,其细菌种类存在一定差别。     

Magnetic orientation in the semi-terrestrial amphipod, Orchestia cavimana, and its interrelationship with photo-orientation and water loss

ABSTRACT. Orchestia cavimana Heller (Amphipoda, Talitridae) were shown to orient to the geomagnetic field as well as to an anisotropic light field. When tested in an isotropic light field the orientation in the geomagnetic field was either in or opposite to the compass direction of the light vector of the anisotropic light field in which the animals had lived before the test, and this orientation was upset predictably by changing the magnetic field with Helmholtz coils. The polarity of the reaction in both magnetic orientation and photo-orientation was correlated with pre-experimental water loss. The magnetic orientation of O. cavimana is compared with that of Tenebrio molitor , and its biological significance discussed.     

Insect growth regulatory effects of some extracts and sterols from Myrtillocactus geometrizans (Cactaceae) against Spodoptera frugiperda and Tenebrio molitor

A methanol extract from the roots and aerial parts of Myrtillocactus geometrizans (Cactaceae) yielded peniocerol 1, macdougallin 2, and chichipegenin 3. The natural products 1, 2 their mixtures, MeOH and CH(2)Cl(2) extracts showed insecticidal and insect growth regulatory activity against fall armyworm [Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae)], an important insect pest of corn, and [Tenebrio molitor (Coleoptera)], a pest of stored grains in Mexico. The most active compounds were 1, 2, and a mixture (M(2)) of 1 and 2 (6:4). All these extracts, compounds and the mixture had insect growth regulating (IGR) activity between 5.0 and 50.0 ppm and insecticidal effects between 50 and 300 ppm in diets. The extracts were insecticidal to larvae, with lethal doses between 100 and 200 ppm. These compounds appear to have selective effects on the pre-emergence metabolism of Coleoptera, because in all treatments of the larvae of T. molitor, pupation were shortened and this process show precociousness in relation to controls. In contrast to S. frugiperda larvae, onset of pupation was noticeably delayed. Emergence in both cases was drastically diminished. In both pupae and in the few adults that were able to emerge, many deformations were observed. The results of these assays indicated that the compounds were more active than other known natural insect growth inhibitors such as gedunin and methanol extracts of Cedrela salvadorensis and Yucca periculosa. Peniocerol, macdougallin and chichipegenin, as well as mixtures of these substances, may be useful as natural insecticidal agents.     

Glutathione S-transferase in the defence against pyrethroids in insects

The correlation between the natural levels of GST and the tolerance to the insecticide decamethrin (dMT), as well as the interaction between the molecules of affinity purified enzyme and the insecticide were investigated in order to collect further information on the obscure role of the Glutathione S-transferase system (GST) as a mechanism of defence against pyrethroids. The studies were carried out, comparatively, on the larvae and pupae developmental stages of the coleopteran Tenebrio molitor, which exhibit varying natural levels of GST activity. No stage dependent susceptibility of the insect against pyrethroid insecticides was found during the first 24 h, however 48 h after treatment, the KD50 dose increased significantly due to the recovery of some individuals from the larvae stage. Simultaneous injection of decamethrin with compounds which inhibit GST activity in vitro, resulted in an increased tolerance, which was more pronounced in the pupae stage. Inhibition studies combined with competitive fluorescence spectroscopy and high pressure liquid chromatography (HPLC) showed that the insecticide binds probably to the active site of the enzyme inhibiting its activity towards CDNB in a competitive manner, but is not conjugated with GSH. According to this, GST offers a passive protection towards pyrethroid insecticides by binding to their molecule in a sequestering mechanism.     

Periodic muscular activity and its possible functions in pupae of Tenebrio molitor

Abstract. During pupal development, Tenebrio molitor L. show regular periods of rhythmic muscular contractions and associated body movements. These periods of activity last 2.5-5.8 min and are more frequent in newly ecdysed pupae ( c. 3h^-1). They become less frequent ( c. 1.5 h^-1) when the basal metabolism reaches its lowest level. In the pharate adult stage the clear pattern of muscular activity disappears.
Muscular activity is temperature-dependent and is commonly absent below 20C. Muscular activity did not disturb the cyclic output of CO₂, which is characteristic of metamorphosis. The heart shows characteristic periods (1–3 min) of activity during pupal development. The frequency of these heart pulsation periods depends on metabolic rate. Heart pumping was correlated mostly with muscular contractions. Therefore we suggest that the main physiological function of muscular activity is to support circulation.     

Interaction of wheat monomeric and dimeric protein inhibitors with alpha-amylase from yellow mealworm (Tenebrio molitor L. larva).

The highly purified alpha-amylase from Tenebrio molitor L. larva (yellow mealworm) reversibly combines with two closely related homogeneous glycoprotein inhibitors, one dimeric (termed 'inhibitor 0.19') and one monomeric (termed 'inhibitor 0.28'), from wheat flour. As established by means of difference spectroscopy and kinetic studies, molar combining ratios for the amylase--inhibitor-0.19 and amylase-inhibitor-0.28 complexes were 1:1 and 1:2 respectively. Two amylase--inhibitor-0.19 complexes with slightly different retention volumes on Bio-Gel P-300 and only one amylase--inhibitor-0.28 complex were observed. Dissociation constants of the amylase--inhibitor-0.19 and amylase--inhibitor-0.28 complexes were 0.85 nM and 0.13 nM respectively. A strong tendency of both complexes to precipitate under an ultracentrifugal field was observed; the minimum molecular weight calculated for the two complexes under such conditions was approx. 95 000. The two complexes showed difference spectra indicating involvement of structurally related or identical tryptophyl side chains in the binding of inhibitors 0.28 and 0.19 to the amylase. A model summarizing the main features of the inhibition of the insect amylase by the two wheat protein inhibitors is proposed.     

Immunocytochemical localization of a diuretic hormone of the beetle Tenebrio molitor,Tenmo-DH(37), in nervous system and midgut

Although the mealworm Tenebrio molitor inhabits very dry environments, it has at least two diuretic peptides, which increase fluid secretion by the free portions of the Malpighian tubules. Unlike other insect corticotropin-releasing factor (CRF)-related peptides isolated to date, these are non-amidated peptides. The immunocytochemical localization of Tenmo-DH(37) was investigated using antisera raised against this hormone. Immunoreactive neurosecretory cells were found in the brain and abdominal ganglia with immunoreactive processes projecting to the peripheral nervous system. Intense staining of the neurohaemal release site, the corpora cardiaca, was observed. In addition, neurosecretory cells immunoreactive to Tenmo-DH(37) were found in the posterior midgut and a network of immunoreactive nerve processes extended over the surface of the midgut. Tenmo-DH(37) is widely distributed and its staining pattern resembles that found for other, amidated CRF-related diuretic peptides.     

饲料蛋白质含量对黄粉虫生长繁殖及川硬皮肿腿蜂寄生能力的影响

【目的】筛选出适宜川硬皮肿腿蜂Scleroderma sichuanensis Xiao寄生的黄粉虫Tenebrio molitor饲料配方。【方法】用10种不同蛋白质含量的饲料饲喂黄粉虫,并观察各虫态生长繁殖情况,再用川硬皮肿腿蜂接种各配方黄粉虫蛹后置于培养箱中培养,5 d后计数雌蜂产卵量,并通过显著性分析、相关性分析及一元回归分析进行数据处理。【结果】各配方黄粉虫生长繁殖情况正常,指标间存在一定的显著性和相关性且与一元回归分析所得相关性一致。川硬皮肿腿蜂对黄粉虫蛹寄生率没有显著性,为86.25%~96.97%,表现出较高寄生率。寄生产卵量较高且蛋白质含量为14.09%和13.73%配方组显著高于其他配方组。【结论】黄粉虫饲料中蛋白质含量在14%(13.73%~14.09%)时川硬皮肿腿蜂寄生能力最强。     

Effect of weak electromagnetic radiation on larva development and metamorphosis of grain beetle Tenebrio molitor

The effect of weak electromagnetic radiation (36 GHz, 100 mu W/cm2) on the development of the grain beetle Tenebrio molitor was studied. Insects were irradiated in different larval instars and at the pupal stage. It was found that weak electromagnetic radiation stimulated the molting and pupation of larvae and the metamorphosis of pupae. The stimulating effect of radiation was weak when animals were exposed in the initial period of the instar and the pupal stage and was more pronounced if the irradiation was carried out in the second half of the current instar and the pupil stage. The effect of weak electromagnetic radiation on the development of beetle can be related to the function of the hormones of metamorphosis.     

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.     

Biological evaluation of analogues of an insect neuropeptide proctolin

Continuing our studies on proctolin (Arg-Tyr-Leu-Pro-Thr) we performed the synthesis and biological evaluation of 52 analogues substituted in position 2, 3, 4, and 5 of the peptide chain. The peptides were bioassayed for cardiotropic activity in vitro on Tenebrio molitor and myotropic activity on foregut of Schistocerca gregaria. Twenty analogues retained 20-80% of proctolin activity.