Connections between adjacent retinula cell columns in the eye of Ephestia kuehniella Zeller (Lepidoptera,Pyralididae)

Summary Connections were found between retinula cells of adjacent retinula cell columns in the eye of Ephestia. The connections occur most frequently at the level of the retinula cell nuclei and may involve two or more retinula columns simultaneously. The absence of specialized structural modifications of the membranes and the presence of pigment granules at the level of the connections or distal to them indicates that these connections are probably not involved in selective chemical or electrical communication nor in light transmission. It is suggested that the connections may serve tactily to coordinate cytoplasmic movement in adjacent retinula columns during light-dark adaptation.Senior U. S. Scientist Awardee of the Alexander von Humboldt Foundation. Thanks to Prof. Dr. A. Fischer for his suggestions and assistance.     

Der Feinbau des Auges der Mehlmotte,Ephestia kuehniella Zeller (Lepidoptera,Pyralididae)

Zusammenfassung Die Retinula im Ommatidium der Mehlmotte besteht aus einer wechselnden Anzahl (9–12, meist 11) langgestreckter, prismatischer Sinneszellen. Außerdem enthält jede Retinula nahe der Basalmembran im Zentrum zwischen diesen distalen Retinulazellen noch eine basale Retinulazelle. Die Längsachse der Retinula wird von der Achsenstruktur eingenommen, die aus Mikrovilli besteht. Ihr distaler Teil ist der Achsenfaden, der breitere, proximale Teil bildet das Rhabdom. Dieses erscheint im Querschnitt meist vierstrahlig gelappt, da seine Außenseite in Längsrichtung tief gekehlt ist. Der Rhabdomquerschnitt gliedert sich in mehrere Schöpfe parallel angeordneter Mikrovilli (Rhabdomsektoren); jeder Rhabdomsektor besteht aus 1 oder 2 Rhabdomeren. Die basale Retinulazelle entsendet einen kleinen Schopf von Mikrovilli in die proximale Spitze des Rhabdoms. Die distalen Retinulazellen setzen sich proximal in Neuriten fort, welche sich in Einkehlungen der basalen Retinulazelle bzw. der Tracheenendzelle einschmiegen. Jeweils eine Tracheole durchbricht zusammen mit dem Neuritenstrang einer Retinula die Basalmembran; sie verzweigt sich distal zu ca. 30 Tracheolen, die die Retinula umhüllen.Die Kristallkegelzellen grenzen distal an die Cornea; proximal laufen die Kristallkegelzellen eines Ommatidiums in einen gemeinsamen Fortsatz aus, der zwischen den Retinulazellen unmittelbar am Achsenfaden endet. — Nur das helladaptierte Auge wurde untersucht. Hierbei erscheint im distalen Teil der Retinula nur der Achsenfaden lichtdurchlässig, das Cytoplasma der Retinulazellen hingegen von Pigmentgrana durchsetzt und für Licht undurchlässig.

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Fine structure of the eye of the meal moth, Ephestia kuehniella Zeller (Lepidoptera, Pyralididae)

Summary In each ommatidium of the meal moth a retinula is formed from a varying number (9–12, mostly 11) of elongated, prismatic sense cells. In addition, a basal retinular cell is situated near the basement membrane in the center of the other (distal) retinular cells. The axis of the retinula is occupied by many microvilli forming the axial structure, the distal section of which is the slender axial thread. Proximally, the axial structure widens (to 8.5 m instead of 1 m in diameter) and is now called rhabdom. Cross sections of the rhabdom mostly look like a petaloid with four petals; this figure is due to longitudinal infoldings along the length of the rhabdom surface. The rhabdom cross section is subdivided into several brushes of microvilli (rhabdom sectors), each one being characterized by an approximately parallel arrangement of its microvilli. One rhabdom sector may be composed of one or two rhabdomeres respectively.The basal retinular cell participates in rhabdom formation through a small brush of microvilli at the proximal end of the rhabdom. Proximally, the distal retinular cells taper into slender neurites which are embedded in grooves at the surface of the basal retinular cell and the tracheal end cell respectively. One tracheole piercing the basement membrane together with the neurites of one retinula branches into about 30 tracheoles surrounding the retinula.The crystalline cone cells touch the cornea; proximally, their cytoplasm forms a point which eventually terminates amongst the distal tips of the retinular cells, immediately at the axial thread.—Our work was restricted to light adapted eyes; in this condition, light transmission in the distal part of the retinula seems to be blocked by retinular cell pigment except inside the axial thread.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

The structure of Verson's cells in Ephestia kuehniella Z. (Pyralidae,Lepidoptera)

Summary Testis follicles of Lepidoptera contain a large somatic cell termed Verson's cell. The present study focuses on the structure of Verson's cells and neighbouring germ cells in the Mediterranean mealmoth, Ephestia kuehniella (Pyralidae), using electron microscopy, antitubulin immunofluorescence, and phalloidin incubation for the visualization of microfilaments. Verson's cells of young larvae are connected with the follicle boundary and show large areas containing packages of glycogen particles, whereas Verson's cells of pupae lie freely within the testis follicle and are largely devoid of glycogen. Both developmental stages of Verson's cells have in common the presence of a dense cytoplasmic network of microtubules. A juxtanuclear subset of the cytoplasmic microtubule array is recognized by an antibody against acetylated microtubules. This indicates that more stable microtubules exist in this region. Microfilaments are arranged parallel to the cytoplasmic microtubules. The microtubule-microfilament-complex forms a cytoskeleton that may keep larger organelles, such as mitochondria and lysosomes, in a juxtanuclear position. Chromatin within the nuclei of Verson's cells is largely decondensed and nuclear pores are abundant. This indicates a high synthetic activity within the cells. The development of cells directly attached to Verson's cells, viz. prespermatogonia, may be controlled by the Verson's cells. Prespermatogonia, which differ in cytoplasmic density from spermatogonia further away from Verson's cells, may represent stem cells that give rise to spermatogonia and somatic cyst cells upon detachment from Verson's cells. This suggestion is compatible with the low division rate of prespermatogonia.     

Effect of diallyl disulfide on physiological performance of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae)

Ephestia kuehniella Zeller is a key pest by severe deficiencies on quantity and quality of stored products. Due to concerns made by synthetic insecticides, use of natural products (like botanical insecticides) is of great importance as an alternative pest management. In the current study, the effect of diallyl disulfide was determined on digestive enzymes and those involved in intermediary metabolism of E. kuehniella. Concentration of 0.31% was obtained as LC₅₀ of diallyl disulfide followed by bioassay experiments. Activities of α-amylase, proteases, alanine aminotransferase, alkaline phosphatase and lactate dehydrogenase significantly decreased in the treated larvae, while the adverse effects were observed in case of triacylglyceride-lipase, aspartate aminotransferase, γ-glutamyl transferase, aldolase and acid phosphatase. Results confirmed that diallyl disulfide interrupted larval digestion by lower activities of digestive enzymes indicating significant effect on intermediary metabolism.     

Five years of mass trapping of Ephestia kuehniella Zeller: a component of IPM in a flour mill

Results obtained by mass trapping using the synthetic pheromone (Z,E)‐9,12‐tetradecadienyl acetate (TDA) are reported. The trapping was carried out over a period of 5 years to control infestation by Mediterranean flour moth, Ephestia kuehniella Zeller, and protect a flour mill. From March 2004 till November 2008, 42 funnel traps, each baited with 2 mg of TDA, were placed in the mill. Eight additional traps were located at the exterior of the facility, especially in the wheat silo area and near the loading equipment. Over 5 years, the pheromone traps attracted a total of 54 170 male Mediterranean flour moths. The constant presence of the traps caused a marked decrease in the E. kuehniella population from 2004 to 2008. The results of the surveys show that the population density of the moth can be notably reduced and then maintained at a low level in flour mills by means of mass trapping techniques accompanied by localized insecticide treatments and careful cleaning of the various departments and of the interior of all equipment. As it is not possible at present to establish the economic injury level of insect infestation in flour mills, there is a need for investigations into how the effectiveness of IPM programs can be evaluated in more detail.     

The restructuring of the flagellar base and the flagellar necklace during spermatogenesis of Ephestia kuehniella Z. (Pyralidae,Lepidoptera)

Summary Transmission electron microscopy was used to study the development of the flagellar base and the flagellar necklace during spermatogenesis in a moth (Ephestia kuehniella Z.). Until mid-pachytene, two basal body pairs without flagella occur per cell. The basal bodies, which contain a cartwheel complex, give rise to four flagella in late prophase I. The cartwheel complex appears to be involved in the nucleation of the central pair of axonemal microtubules. In spermatids, there is one basal body; this is attached to a flagellum. At this stage, the nine microtubular triplets of the basal body do not terminate at the same proximal level. The juxtanuclear triplets are shifted distally relative to the triplets distant from the nuclear envelope. Transition fibrils and a flagellar necklace are formed at the onset of axoneme elongation. The flagellar necklace includes Y-shaped elements that connect the flagellar membrane and the axonemal doublets. In spindle-containing spermatocytes, the flagellar necklace is no longer detectable. During spermatid differentiation, the transition fibrils move distally along the axoneme and a prominent middle piece appears. Our observations and those in the literature indicate certain trends in sperm structure. In sperms with a short middle piece, we expect the presence of a flagellar necklace. The distal movement of the transition fibrils or equivalent structures is prevented by the presence of radial linkers between the flagellar membrane and the axonemal doublets. On the other hand, the absence of a flagellar necklace at the initiation of spermiogenesis enables the formation of a long middle piece. Thus, in spermatozoa possessing an extended middle piece, a flagellar necklace may be missing.     

Ultrastructure of distal flagellar swellings in spermatocytes and spermatids of Ephestia kuehniella Z. (Pyralidae,Lepidoptera)

Summary Development of flagella was investigated by transmission electron microscopy in spermatocytes and spermatids of the Mediterranean mealmoth, Ephestia kuehniella Z. Growing flagella displayed voluminous distal swellings. In short flagella the apical portion of the swellings contained an amorphous, dense accumulation. In more developed flagella a less dense proximal extension of the apical accumulation was formed, which in turn was in contact with the elongating flagellar microtubules. The material of the flagellar tip is interpreted as being a precursor of the axoneme containing mainly tubulin. The material may be converted into the axoneme.     

Tolerance to Bacillus thuringiensis endotoxin in immune-suppressed larvae of the flour moth Ephestia kuehniella

Tolerance to Bacillus thuringiensis crystal endotoxins (Bt-toxins) is correlated with an elevated immune status in larvae of the flour moth Ephestia kuehniella. To gain more specific information about the effector pathways involved in the protection against the toxin, we studied the effects of Bt-toxin formulations in susceptible (non-induced) and tolerant (immune-induced) larvae after natural (parasitism-mediated) and chemical (tropolone-mediated) suppression of defence reactions. Although melanization in hemolymph was significantly reduced, there was no significant effect on susceptibility to the toxin in parasitised or tropolone-treated larvae. This suggests that melanization of hemolymph is correlated with an elevated immune status but not responsible for the observed tolerance to Bt-toxin. To examine whether hemolymph proteins exist in the gut lumen and function as pro-coagulants, we compared gut and plasma proteins of immune-induced with those of non-induced larvae. Here we show that the lipid carrier lipophorin represents a major component in the gut lumen and interacts with mature Bt-toxin to form a complex.     

Infection density of Wolbachia and level of cytoplasmic incompatibility in the Mediterranean flour moth,Ephestia kuehniella

Wolbachia, a causative agent of various reproductive changes in arthropods, induces cytoplasmic incompatibility (CI) in the Mediterranean flour moth, Ephestia kuehniella. Two strains of E. kuehniella, Yokohama and Tsuchiura, harbor closely related Wolbachia, but the Yokohama strain expresses stronger CI than the Tsuchiura strain. A transinfected E. kuehniella strain that harbors the Wolbachia derived from the almond moth Cadra cautella, expresses weak CI at a similar level to the Tsuchiura strain. In the present study, we measured the Wolbachia density in the testis of the three E. kuehniella strains in order to examine the effects of bacterial strain and infection load on the expression of CI. When individuals of the same strain were compared, a correlation of bacterial density to CI level was observed. In addition, the Wolbachia density was higher in the Yokohama strain than the Tsuchiura strain in agreement with the CI levels expressed. However, this relationship did not hold in the comparison between the naturally infected and transinfected strains that carried phylogenetically distant Wolbachia.     

Genetic variation in progeny allocation in Trichogramma maidis

A considerable amount of work has been recently devoted to the study of progeny allocation strategies by insect parasitoids under different environmental constraints (see Waage, 1986, for a review). All these studies present each of the oviposition strategies (e.g. number of eggs laid per host, superparasitism) as a phenomenon which is progressively settled by natural selection during the course of generations. However, this optimal progeny allocation theory is conceivable only if the variation in wasp's biological traits involved are under a genetic control on which natural selection could act. We have decided therefore to look for a genetic variability in these traits in a Trichogramma maidis Pintureau & Voegelé (Hym.; Trichogrammatidae) population.Based on histological staining which allows to count the number of Trichogramma eggs deposited in each host (Ephestia kuehniella Zeller), we compared different wasp isofemale lines (i.e. families) each of which was submitted to different conditions (i.e. parasite/host ratios).The results show that the control of superparasitism seems to be genetically determined. This appears true both for the average number of eggs deposited in each host, and for the frequency distribution of wasp eggs among hosts.The evolutionary consequences of such results and their possible applications in order to improve the wasp's mass-rearing conditions are discussed.

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Résumé Récemment, de nombreux travaux ont été publiés dans le but de mieux comprendre les différentes stratégies de reproduction (i.e. répartition de la progéniture parmi les hôtes) présentées par les insectes parasitoides sous différentes contraintes environnementales (cf. Waage, 1986, pour une synthèse). L'ensemble de ces travaux considère chacune de ces stratégies (e.g. nombre d'ufs déposés par hôte, décision de superparasiter) comme un phénomène progressivement mis en place par la sélection naturelle au cours des générations. Ceci n'est cependant concevable que si les caractères biologiques en cause chez les parasites sont sous la dépendance d'une variation génétique sur laquelle la sélection naturelle peut agir.Les expériences présentées ici ont donc eu pour but de voir si ces caractères sont effectivement déterminés génétiquement dans une population de Trichogramma maidis Pintureau & Voegelé (Hym.; Trichogrammatidae).Grâce à l'utilisation d'une coloration histologique qui permet de compter le nombre d'ufs de trichogrammes déposés dans chaque hôte, différentes lignées isofemelles (i.e. familles) ont été comparées, et ce selon différentes conditions expérimentales correspondant à différentes valeurs du rapport nombre de femelles parasites sur nombre d'hôtes offerts.Les résultats obtenus motrent que le phénomène du superparasitisme semble être déterminé génétiquement. Ceci est constaté aussi bien au niveau du nombre moyen d'ufs de trichograme déposés par hôte, que dans les caractéristiques des distributions des ufs de parasites parmi les hôtes.Les conséquences évolutives de tels résultats, ainsi que leurs applications possibles dans le but d'améliorer les conditions d'élevage de masse de ces insectes, sont discutées.

     

Investigation of the steps involved in the difference of susceptibility of Ephestia kuehniella and Spodoptera littoralis to the Bacillus thuringiensis Vip3Aa16 toxin

BUPM95 is a Bacillusthuringiensis subsp. kurstaki strain producing the Vip3Aa16 toxin with an interesting insecticidal activity against the Lepidopteran larvae Ephestia kuehniella. Study of different steps in the mode of action of this Vegetative Insecticidal Protein on the Mediterranean flour moth (E. kuehniella) was carried out in the aim to investigate the origin of the higher susceptibility of this insect to Vip3Aa16 toxin compared to that of the Egyptian cotton leaf worm Spodoptera littoralis. Using E. kuehniella gut juice, protoxin proteolysis generated a major band corresponding to the active toxin and another band of about 22 kDa, whereas the activation of Vip3Aa16 by S. littoralis gut juice proteases generated less amount of the 62 kDa active form and three other proteolysis products. As demonstrated by zymogram analysis, the difference in proteolysis products was due to the variability of proteases in the two gut juices larvae. The study of the interaction of E. kuehniella BBMV with biotinylated Vip3Aa16 showed that this toxin bound to a putative receptor of 65 kDa compared to the 55 and 100 kDa receptors recognized in S. littoralis BBMV. The histopathological observations demonstrated similar damage caused by the toxin in the two larvae midguts. These results demonstrate that the step of activation, mainly, is at the origin of the difference of susceptibility of these two larvae towards B. thuringiensis Vip3Aa16 toxin.     

Effect of temperature on survival, development and reproduction of the predatory lacewing Dichochrysa prasina (Neuroptera: Chrysopidae) reared on Ephestia kuehniella eggs (Lepidoptera: Pyralidae)

Preimaginal development and adult longevity and reproduction of Dichochrysa prasina Burmeister were studied at six constant temperatures (15, 20, 25, 27, 30 and 33 °C) and a photoperiod of 16:8 (L:D). Eggs of the flour moth Ephestia kuehniella (Zeller) were used as food throughout preimaginal development, whereas the adults of D. prasina fed on a liquid diet of water, yeast hydrolysate, sugar and honey. At the highest tested temperature of 33 °C no larvae completed their development. At the rest of the tested temperatures the egg to adult developmental period ranged from approximately 92 days at 15 °C to 25 days at 30 °C. Percentages of adult emergence ranged from 36% at 15 °C to 84% at 30 °C. Both adult longevity and fecundity were significantly affected by temperature and the intrinsic rate of increase (r_m) reached its maximum value at 27 °C. These results could be useful for the establishment of a small scale rearing and mass production of D. prasina.     

Assessment of quality control parameters during mass rearing of a pupal colour genetic sexing strain of the Mediterranean fruit fly

Quality control charts were employed to monitor the quality of several parameters during mass rearing of the T : Y(wp ⁺)30C strain of Ceratitis capitata (Wiedemann), in which male and female flies emerge from brown and white puparia, respectively. Egg hatch and pupal recovery in this genetic sexing strain were below that reported for a wild-type Sohag strain, whereas pupal size, adult emergence and flight ability were higher. The overall recovery from eggs to adult flies capable of flight was similar in the two strains but egg production in T:Y(wp ⁺)30C was reduced to about 56% of that of Sohag. The quality control charts showed an acceptable level of variation within sets of data but the level between data sets was occasionally out of control. This indicated the presence of uncontrolled conditions during the rearing process, possibly arising from variations in the quality of the larval diet. It is concluded that the quality of T : Y(wp ⁺)30C renders this strain suitable for mass rearing in sterile insect technique (SIT) programmes requiring the elimination of the female flies prior to the release of sterilized males. The next step should involve further quality assessments both under field cage and under field release conditions. The importance of optimal and standardized dietary conditions is stressed in relation to the quality of mass reared medflies.

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Résumé Des normes de qualité ont été utilisées pour relever la valuer de plusieurs paramètres lors de l'élevage industriel de la souche T : Y(wp ⁺)30C de C. capitata Wied. dont les mâles et les femelles émergent respectivement de pupes brunes et blanches. Les taux d'éclosion et d'obtention de pupes sont plus faibles avec cette souche qu'avec la souche souvage Sohag; par contre, la taille des pupes, le taux d'émergence des adultes et l'aptitude au vol étaient plus élevés. La fréquence totale des adultes capables de voler par rapport au nombre initial d'ufs a été de 0,297 contre environ 0,311 pour Sohag. Cependant, la production d'ufs a été réduite à environ 56% de celle de Sohag. Les tableaux de contrôle de la qualité ont révélé un niveau de variation acceptable à l'interieur rieur des lots de données, mais la variabilité entre les lots excédait les normes admissibles. Ceci implique des paramètres incontrôlés pendant l'élevage de masse, probablement liés à des variations de la qualité de l'aliment. On peut en conclure que la qualité de la souche T : Y(wp ⁺)30C la rend utilisable pour un élevage de masse dans des programmes d'utilisation d'insectes stériles (SIT) exigeant l'élimination des femelles avant le lâcher de mâles stériles. La prochaine étape impliquera de nouvelles épreuves de qualité dans des cages en champ et dans les conditions de libération dans la nature. L'importance des conditions optimales d'une alimentation standardisée est soulignée par la qualité des mouches obtenues en élevage de masse.

     

Heterosis in a wild strain of Drosophila polymorpha with a lethal closely linked to the major esterase locus

A laboratory population derived from a single wild inseminated female has a lethal so closely linked to the major esterase locus that in 3 years of observations no crossover products have been detected. Linkage with a chromosome inversion was excluded by cytological analysis. The heterozygotes are superior to the homozygotes in egg-adult viability, egg-laying rate, and longevity. Electrophoretic analysis of larvae, pupae, and 2-h-old adults shows that the specific phase for the lethal effect in homozygotes is the pupal stage.     

Genetic variability in the reactive distance inTrichogramma brassicae after automatic tracking of the walking path

Efficiency of hymenopterous parasitoids to control pests in a biological control release program probably increases with increasing searching ability of the females, which, in return, likely depends on the distance from which females perceive their hosts (i.e., reactive distance). In this study, we first analyse this hypothesis with the help of a stochastic model simulating the walking path of isolatedTrichogramma females during their searching behaviour. Then, this reactive distance is estimated using automatic recording and analysis of the walking path of female wasps. Finally, the genetic variability for this trait is analysed in aTrichogramma brassicae Bezdenko (Hym.; Trichogrammatidae) population. Three types of hosts were used: Eggs ofEphestia kuehniella Zeller (Lep.; Pyralidae), eggs ofMamestra brassicae L. (Lep.; Noctuidae) and 0.45 mm glass beads.M. brassicae eggs are perceived from 4.01±0.15 mm, which is a significantly longer perceptive distance than forE. kuehniella eggs (3.69±0.10 mm) and glass beads (3.67±0.10 mm). Moreover, whatever the host tested, a significant genetic variation is observed in this trait in the population studied. The ecological and evolutionary implications and the agronomical importance of these results are discussed.     

Genetic control of lactate dehydrogenase isozymes in Paramecium caudatum

The LDH isozymes were surveyed in bacterized cultures of syngens 1, 3, 12, and 13 of Paramecium caudatum by polyacrylamide gel electrophoresis. All the examined stocks of different syngens except one stock in syngen 3 had a single common LDH isozyme, and intra- and intersyngenic variation was not observed except for the one stock in syngen 3. Breeding data using the exceptional stock indicated that the LDH isozymes of P. caudatum are controlled by two codominant alleles at a single locus whose products aggregate randomly, forming a dimer.     

Introduction and establishment of parasitoids for the biological control of the apple ermine moth, Yponomeuta malinellus (Lepidoptera: Yponomeutidae), in the Pacific Northwest

Four parasitoids were imported from five countries in Eurasia and released in northwestern Washington, US, to control the apple ermine moth (AEM), Yponomeuta malinellus Zeller, which colonized the Northwest around 1981. From 1988 to 1991, 95,474 individuals of Ageniaspis fuscicollis (Dalman) from France, China, Korea, and Russia were released in Washington. Parasitism of AEM increased 4- to 5-fold over that produced by preexisting natural enemies between 1989 and 1994 at 22 monitored sites. Subsequently, the wasp dispersed up to 20 km from release sites. A. fuscicollis also parasitized the cherry ermine moth, Yponomeuta padellus (L.), which was discovered in the Pacific Northwest in 1993. A total of 1813 individuals of Herpestomus brunnicornis (Gravenhorst) from France, Korea, and Japan were released in 1989–1991, and 26 wasps were recovered in 1994–1995. From 1989 to 1991, 2647 Diadegma armillata (Gravenhorst) individuals from France were released. D. armillata was recovered at one site in 1991 two months following release, but no other recoveries have been made. A total of 8274 Eurystheae scutellaris(Robineau-Desvoidy) individuals were released in 1989 to 1991. However, this tachinid has not been recovered. A consistent decline of AEM populations occurred in 1989–1995, including at sites where A. fuscicollis was not recovered, suggesting other factors also contributed to this pest’s decline. Now well established in western Washington, A. fuscicollis may help suppress future outbreaks of Y. malinellus and its congener, Y. padellus.