Asymmetric larval head and mandibles of Hydrophilus acuminatus (Insecta: Coleoptera, Hydrophilidae): Fine structure and embryonic development.

The larvae of a water scavenger beetle, Hydrophilus acuminatus, have strongly asymmetric mandibles; the right one is long and slender, whereas the left one is short and stout. The fine structure and embryonic development of the head capsule and mandibles of this species were examined using light and scanning electron microscopy, and asymmetries in shape were detected in these structures applying an elliptic Fourier analysis. The larval mandibles are asymmetric in the following aspects: whole length, the number, structure and arrangement of retinacula (inner teeth), and size and shape of both the molar and incisor regions. The larval head is also asymmetric; the left half of the head capsule is larger than the right, and the left adductor muscle of the mandible is much thicker than the right. The origin and developmental process of asymmetric mandibles were traced in developing embryos whose developmental period is about 270 h and divided into 10 stages. Mandibular asymmetries are produced by the cumulative effects of six stepwise modifications that occur from about 36% of the total developmental time onward. The significance of these modifications was discussed with respect to the functional advantages of asymmetries and the phylogeny of members of the Hydrophilidae.

Predaceous diving beetle, Dytiscus sharpi sharpi (Coleoptera: Dytiscidae) larvae avoid cannibalism by recognizing prey.

Larvae of diving beetles such as the various Dytiscus species (Coleoptera: Dytiscidae) are carnivorous and usually prey on other aquatic animals. Cannibalism among larvae of Dytiscus sharpi sharpi (Wehncke) was observed to begin when they were starved for more than two days under artificial breeding conditions. However, the 2-day starved larvae did not show cannibalism in the presence of intact, motionless, frozen tadpoles, or frozen shrimps. The beetle larvae attacked and captured intact tadpoles faster (15 sec) than other motionless and frozen tadpoles (120 sec), indicating that prey movement was an important factor in stimulating feeding behavior in larvae. Prey density does not have an effect on larval cannibalism. In cases in which preys are present at lower densities than that of larvae, a group of beetle larvae frequently fed on single prey. This feeding behavior, therefore, provides direct evidence of self-other recognition at the species level. Using two traps in one aquarium that allows the larvae to detect only prey smell, one containing tadpoles and another empty, the beetle larvae were attracted to the trap with tadpoles at high frequency, but not to the empty trap. In another experiment, the beetle larvae were not attracted to the trap containing a beetle larva. These results suggest that the larvae of D. sharpi sharpi are capable of recognizing prey scent, which enables the promotion of foraging behavior and the prevention of cannibalism.

The inheritance of intrasexual dimorphism in female diving beetles (Coleoptera: Dytiscidae).

Many species of Dytiscus diving beetles exhibit intrasexual dimorphism, e.g., the elytra is smooth in some females and grooved in others. However, the expression of the grooves and whether they are a product of heredity or the environment remain unknown. One Japanese species, Dytiscus sharpi sharpi Wehncke, 1875 , also shows female dimorphism, with grooved and smooth morphs, while D. sharpi validus Régimbart, 1899, only has a single morph (the grooved type). A hybrid of the two species should therefore provide a means of sorting out how the grooves are inherited. We found two independent wetlands of D. sharpi sharpi in Chiba Prefecture, Japan. One was a place where a high proportion of grooved females lived, and the others had high proportions of smooth females. After five to eight generations of beetles from two populations with different proportions of grooved females were reared under aquarium conditions constituting a common garden design, i.e., water temperature, water depth, and presence of a plant for oviposition, the differences remained. We mated smooth virgin females of D. sharpi sharpi with males of D. sharpi validus to obtain hybrid offspring. The elytral traits of the hybrid females produced only grooved forms. These results suggested that the female dimorphism is determined by genetics, and that the grooved morph was dominant over the smooth one, independent of environmental factors. In addition, the hybrid insects did not differ from the two subspecies insects in larval survivorship, pupation success, or sex ratio. They also showed neither morphological abnormality nor reduced survival.

Temperature-dependent regulation of reproduction in the diving beetle Dytiscus sharpi (Coleoptera: Dytiscidae).

The effects of temperature on the mating behavior, gonad development, germ cell maturation, and egg spawning of the predaceous diving beetle Dytiscus sharpi (Coleoptera; Dytiscidae), were investigated. By field observations, we found that mating behavior started in October and occurred more frequently from November to December. Under our laboratory breeding conditions, we observed almost the same seasonal variation in mating behavior. We found that temperatures lower than 20 degrees C were required to trigger mating behavior. We also found the same temperature threshold triggered gonadogenesis as well as spermatogenesis. Furthermore, for females, exposure to lower temperatures (<8 degrees C) during the winter was required for egg maturation and spawning in spring; that is, there was a second threshold for successful female reproduction. We conclude that the termination of summer reproductive diapause of D. sharpi is regulated in a temperature-dependent manner, thus effecting the adaptation of D. sharpi to southern warm habitats.

Functional characterization of single nucleotide polymorphisms with amino acid substitution in CYP1A2, CYP2A6, and CYP2B6 found in the Japanese population.

As a part of the studies conducted by the Pharma SNPs Consortium (PSC), the enzyme activities of CYP1A2, CYP2A6 and CYP2B6 variants with altered amino acids as a result of single nucleotide polymorphisms (SNPs) found among the Japanese population were analyzed under a unified protocol using the same lots of reagents by the laboratories participating in the PSC. Mutations in CYP1A2, CYP2A6 and CYP2B6 were introduced by site-directed mutagenesis and the wild type and mutated CYP molecules were expressed in Escherichia coli. The expressed cytochrome P450s were purified and the enzyme activities were measured in reconstitution systems. CYP1A2 and CYP1A2Gln478His did not show any differences in 7-ethoxyresorufin O-deethylase activity. CYP2A6 and CYP2A6Glu419Asp metabolized coumarin to form 7-hydroxycoumarin in a similar manner, whereas CYP2A6Ile471Thr showed low activity compared to the wild-type CYP2A6. CYP2B6, CYP2B6Pro167Ala and CYP2B6Arg487Cys showed the same activity for 7-ethoxy-4-triflouromethyl-coumarin O-deethylation. However, CYP2B6Gln172His was roughly twice as active as CYP2B6 and the other CYP2B6 variants for 7-ethoxy-4-triflouromethylcoumarin O-deethylation activity. Although higher inter- and intra-laboratory variations were observed for the calculated Km and V(max) values because the studies were conducted in several different laboratories, the degree of variations was reduced by the increased number of analyses and the adoption of a simple analysis system.

Mating and reproduction of predaceous diving beetles, Dytiscus sharpi, observed under artificial breeding conditions.

Mating season and embryonic development of the predaceous diving beetles, Dytiscus sharpi, (Coleoptera; Dytiscidae) were observed under artificial breeding conditions. Female and male adult insects started mating from November to March and gave first instar larvae mainly in April. When the mating was artificially delayed until February, first instar larvae appeared from the end of March to the middle of May. I also investigated the effects of temperature on larval development. Apparent hatchability of eggs was not affected by high temperature, however, their normal development after hatching was significantly interfered. Most of the first instar larvae kept at 20-25 degrees C from before hatching died within one day after hatching. By contrast, juveniles kept outdoors (7.0-20.9 degrees C) could develop at least until second instar larvae. Temperature >23 degrees C after hatching had no effects on larval development. From these observations, it was concluded that the reproduction strategy of Dytiscus sharpi, i.e. mating in late autumn and hatching in early spring would be the reasonable results of adaptation to the warm habitats where they are collected.