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.     

Spermatophore production and spermatheca content in Tenebrio molitor infected with Hymenolepis diminuta

Male and female Tenebrio molitor act as intermediate hosts for metacestodes of the rat tapeworm, Hymenolepis diminuta. It is known that the bean-shaped accessory glands of infected males exhibit an extended growth period and are significantly larger than those from noninfected males by day 10 after emergence. We wished to determine whether more material is transferred from these glands into the spermatophores. Here we report that the protein content and trehalase activity of spermatophores produced by bean-shaped accessory glands from infected males is elevated. However, protein transferred to the female spermatheca during mating was not affected by the infection status of the male. No evidence of transfer of trehalase to the spermatheca was detected but spermatheca from virgin, infected females contained significantly greater trehalase activity than those from noninfected females.     

Effect of adrenocorticotropin fragments on the memory of Tenebrio molitor beetles

Fragments of the adrenocorticotropic hormone ACTH1-24 and ACTH5-7 facilitate memory in the beetle T. molitor, the effect being observed at different stages after learning. An analogue of ACTH5-7 which contains D-phenylalamine, as well as D-phen7-ACTH4-7, inhibit memorization (the results checked 1 day after learning) and induce some other disturbances in the behaviour of beetles. To the 10th day of learning, the effects of the analogues cannot be distinguished from those of L-Phe7-fragments. The results obtained are compared with the known effect of ACTH fragments on memory in vertebrates.     

On the visually evoked head nystagmus of Tenebrio molitor and other beetles

Head movements of different species of walking beetles elicited by rotating stripe patterns have been investigated. They are of the usual type in contrast to an inverted nystagmus reported forTenebrio molitor in similar experimental situations. Reexamination of theTenebrio records revealed that the sign of the stimulus was interchanged by a mistake while plotting the results. Thus, the head nystagmus inTenebrio is also of the usual type, consisting of a smooth pursuit head movement followed by a faster returning phase.     

Apoptosis in the fat body tissue of the beetle Tenebrio molitor parasitised by Hymenolepis diminuta

Many insects experience a decrease in reproductive output when parasitised. We are investigating mechanisms underlying this fecundity reduction using the rat tapeworm, Hymenolepis diminuta infection of Tenebrio molitor beetles. These include an increase in the resorption of developing ovarian follicles and a decrease in fat body synthesis of vitellogenin. The latter is the direct effect of a molecule produced by the parasite. Here we report a study to determine whether vitellogenin synthesis and follicle resorption are the result of parasite-induced apoptosis in the respective tissues and whether the parasite molecule acts directly on the fat body by inducing apoptosis. In vivo, the number of fat body cell nuclei with chromatin condensation are significantly elevated in parasitised females at all days examined and peaked at day 7 post-infection. A TUNEL assay to detect DNA fragmentation confirmed these observations of apoptosis. However, when fat body from uninfected females was co-cultured with live metacestodes they did not cause cells to die by apoptosis, showing that the induction signal does not come directly from the parasite. The follicle resorption observed in the ovaries of infected beetles was not associated with apoptosis of the epithelial cells. The possibility of several mechanisms underlying fecundity reduction is discussed.     

Klinokinetic and klinotactic humidity reactions of the beetles Hylobius abietis and Tenebrio molitor

ABSTRACT. The effects of spatially uniform but temporally changing air humidity stimuli on the orientation behaviour of the large pine weevil ( Hylobius abietis L.) and the flour beetle ( Tenebrio molitor L.) were studied in a specially constructed vertical air flow chamber. Moist-acclimatized but hygronegative pine weevils were exposed for 10 s to a moist air stream ( c. 85% R.H.) and then for 10 s to a dry air stream ( c. 50% R.H.). The Tenebrio were divided into two groups acclimatized to moist and dry air respectively. For each group the humidity during the initial 5 s in the air stream was the same as during acclimatization, and then changed to the alternative level. In Hylobius the change from moist to dry air caused a decrease in spatial displacement brought about by a decrease in walking speed and an increase in the amount of turning per unit time. In Tenebrio the change from dry to moist air caused a decrease of walking speed and a turning-back reaction based, presumptively, on idiothetic information about the insect's previous walking direction. The average angle turned during 2 s after the humidity change was 165°. In moist-adapted beetles the dry air stream caused an identical orthokinetic arrestment, but no klinokinetic or klinotactic reaction. Klinokinesis is redefined as a change in the circular variance of turning angles per unit time, which does not change the mean direction (or directions in cases of multimodal distributions) of the circular distribution. Accordingly, klinotaxis consists of a change in the mean directions) of the circular distribution of angular velocities. In both species the turning frequency was more constant than turning per unit distance, suggesting a temporal control in the nervous system of klinokinetic and klinotactic behaviour.     

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.     

Structure-activity relationships for the cardiotropic action of the Led-NPF-I peptide in the beetles Tenebrio molitor and Zophobas atratus.

We have examined the effects of the Led-NPF-I peptide (Ala-Arg-Gly-Pro-Gln-Leu-Arg-Leu-Arg-Phe-amide) and a series of ten analogues on the heart contractile activity of Tenebrio molitor and Zophobas atratus, and the structure-activity relationships for cardioactive action of Led-NPF-I were established. A video microscopy technique and computer-based method of data acquisition and analysis were used to study the action of the peptides on continuously perfused heart preparations. Cardiac activity was progressively inhibited by Led-NPF-I when the peptide concentrations were increased from 10(-9) to 10(-5) M. Substitution of the L-proline residue at position 4 of the native peptide with hydroxyproline, valine or D-proline caused a loss of cardioinhibitory activity. Also, replacement of arginine residues at all three positions 2, 7 and 9 with another basic amino acid histidine, reduces cardioinhibitory action of Led-NPF-I. Some modifications of the C-terminal residues, as the Phe(4-NO2)-, Phe(4-NH2)- and Phe(4-NMe2)-analogues, resulted in agonistic peptides with biological activity similar to that of the native peptide. However, three other C-terminal analogues tested [Tyr10]-, [D-Phe10]-Led-NPF-I, and Ala-Arg-Gly-Pro-Gln-Leu-Arg-Leu-Arg-Phe-OH were inactive in the heart bioassay, which suggests that this end of the amino acid chain may play an important role in bioactivity and interaction of the native peptide with its receptor on the myocardium.     

Diversity of digestive proteinases in Tenebrio molitor (Coleoptera: Tenebrionidae) larvae

The spectrum of Tenebrio molitor larval digestive proteinases was studied in the context of the spatial organization of protein digestion in the midgut. The pH of midgut contents increased from 5.2-5.6 to 7.8-8.2 from the anterior to the posterior. This pH gradient was reflected in the pH optima of the total proteolytic activity, 5.2 in the anterior and 9.0 in the posterior midgut. When measured at the pH and reducing conditions characteristic of each midgut section, 64% of the total proteolytic activity was in the anterior and 36% in the posterior midgut. In the anterior midgut, two-thirds of the total activity was due to cysteine proteinases, whereas the rest was from serine proteinases. In contrast, most (76%) of the proteolytic activity in the posterior midgut was from serine proteinases. Cysteine proteinases from the anterior were represented by a group of anionic fractions with similar electrophoretic mobility. Trypsin-like activity was predominant in the posterior midgut and was due to one cationic and three anionic proteinases. Chymotrypsin-like proteinases also were prominent in the posterior midgut and consisted of one cationic and four anionic proteinases, four with an extended binding site. Latent proteinase activity was detected in each midgut section. These data support a complex system of protein digestion, and the correlation of proteinase activity and pH indicates a physiological mechanism of enzyme regulation in the gut.     

Management of entomopathogenic fungi in cultures of Tenebrio molitor (Coleoptera: Tenebrionidae)

Larvae of mealworms Tenebrio molitor L. (Coleoptera: Tenebrionidae) have been used as animal feed, but fungal pathogens rapidly downsize the populations, resulting in economic losses. In this work, we established an effective management strategy for fungal pathogens. An entomopathogenic fungus, Beauveria bassiana, was isolated from mealworm cadavers. The bioassay of some isolates of this species at >90% relative humidity revealed that the ERL1575 isolate had the highest virulence. At 20–30% RH, ERL1575 conidia when ingested produced 80% mortality but when sprayed topically produced only <10% mortality. Mealworms that had ingested conidia were exposed to 20, 25, 30 and 35°C and high humidity (>95%) for 5 days. This experiment produced about 90% mortality except at 35°C where mortality was <20%. When 40 fungicides were assayed against ERL1575, fluazinam (1000‐fold) and mancozeb (667‐fold) significantly inhibited conidial germination and/or hyphal growth. When fluazinam and mancozeb were added to the mealworm diet of conidia‐inoculated wheat bran, most were alive 3 days post application. However, 100% mortality resulted 3 days post application in the conidia‐inoculated wheat bran without any fungicides. In conclusion, B. bassiana isolates are pathogenic at <30°C when they are ingested by mealworms but fluazinam and mancozeb can be used for management to control the pathogen in their cultures.     

A nuptially transmitted ichthyosporean symbiont of Tenebrio molitor (Coleoptera: Tenebrionidae)

The yellow mealworm, Tenebrio molitor, harbors a symbiont that has spores with a thick, laminated wall and infects the fat body and ventral nerve chord of adult and larval beetles. In adult males, there is heavy infection of the epithelial cells of the testes and between testes lobes with occasional penetration of the lobes. Spores are enveloped in the spermatophores when they are formed at the time of mating and transferred to the female's bursa copulatrix. Infection has not been found in the ovaries. The sequence of the nuclear small subunit rDNA indicates that the symbiont is a member of the Ichthyosporea, a class of protists near the animal-fungi divergence.