Darwin in his On the Origin of species made it clear that evolutionary change depends on the combined action of two different causes, the first being the origin of genetically based phenotypic variation in the individual organisms comprising the population and the second being the action of selective agents of the external environment placing demands on the individual organisms. For over a century following Darwin, most evolutionists focused on the origin of inherited variation and its transmission; many workers continue to regard genetics to be the core of evolutionary theory. Far less attention has been given to the exact nature of the selective agents with most evolutionists still treating this cause imprecisely to the detriment of our understanding of both nomological and historical evolutionary theory.

Darwin was vague in the meaning of his new concept of “Natural Selection,” using it interchangeably as one of the causes for evolutionary change and as the final outcome (= evolutionary change). In 1930, natural selection was defined clearly as “non-random, differential reproduction of genes” by R. Fisher and J.B.S. Haldane which is a statement of the outcome of evolutionary process and which omits mention of the causes bringing about this change. Evolutionists quickly accepted this outcome definition of natural selection, and have used interchangeably selection both as a cause and as the result of evolutionary change, causing great confusion. Herein, the details will be discussed of how the external environment (i.e., the environment-phenotype interaction) serves as selective agents and exerts demands on the phenotypic organisms. Included are the concepts of fitness and of the components of fitness (= adaptations) which are respectively (a) survival, (b) direct reproductive and (c) indirect reproductive features. Finally, it will be argued that historical-narrative analyses of organisms, including classification and phylogenetic history, are possible only with a full understanding of nomological evolutionary theory and with functional/adaptive studies of the employed taxonomic features in addition to the standard comparative investigations.

Morphological and functional organization of ON and OFF pathways in the adult newt retina were examined by intracellular recording and staining techniques and immunohistochemistry. Synaptotagmin immunoreactivity discriminated three broad bands within the IPL: the distal band (sublamina I), the middle band (sublamina II) consisting of two dense punctate bands (sublaminae II_a and II_b), and proximal band (sublamina III). The Lucifer-yellow labeled OFF amacrine and ganglion cells send their processes mainly in sublamina I and/or II_a where OFF bipolar cells extend their axon terminals, while ON amacrine and ganglion cells send their processes in sublamina III and/or II_b where ON bipolar cells extend their axon terminals. Processes of ON-OFF amacrine and ganglion cells ramify broadly in the whole thickness of the IPL. Many bipolar cells responded to light spot with a transient hyperpolarization at both light onset and offset. They are probably subtypes of ON bipolar cells, because their axon terminals branch mainly in sublaminae III and/or II_b, although a few cells ramified the axon at both sublaminae II_a and III. Two immunohistochemical markers for bipolar cells, PKC and RB-1, identified axon terminals in sublaminae III and/or II_b. From the ramification pattern of axon terminal, they are probably subtypes of ON bipolar cells. ChAT-ir amacrine cells ramified their dendrites in either sublamina I or II_b. Altogether, present studies support the general idea of segregation of ON and OFF pathways in sublaminae a and b of the IPL.

Serotonin (5-HT) suppresses the photo-responsiveness of medulla bilateral neurons (MBNs) that are involved in the coupling mechanism of the bilaterally paired optic lobe circadian pacemakers in the cricket, Gryllus bimaculatus. We found that forskolin, a highly specific activator of adenylate cyclase, mimicked the effects of serotonin on the MBNs. This fact suggests the involvement of cyclic 3′, 5′-adenosine monophosphate (cAMP) in mediating the action of serotonin. We therefore tested the effects of various 5-HT receptor agonists and antagonists that are coupled to adenylate cyclase to specify the receptor involved. Application of 8-OH-DPAT that has affinity for both 5-HT_1A and 5-HT₇ receptors suppressed the photo-responsiveness, like forskolin. The inhibitory effect of 8-OH-DPAT was effectively blocked by cloza-pine, a high affinity 5-HT₇ receptor antagonists with a very low affinity for 5-HT₂. Ketanserin, a selective 5-HT₂ antagonist, and NAN-190, a 5-HT_1A antagonist, did not block it. These results suggest that serotonergic suppression of the photo-responsiveness of the MBNs is mediated by 5-HT₇-like receptor sub-types.

Foraging flights have been studied in three species of hornets (Vespa mandarinia, V. simillima and V. analis) in the field and the laboratory.

Hornets seem to use multiple navigational cues for visiting a familiar feeding place. They could orient towards the feeding place immediately after they rose in air from the nest without directly viewing the feeder. They could visit the feeding place after dark at a luminosity 8 lux. These data suggest that they can navigate for some distance with few external cues. Hornets also seem to rely on visual cues for their mid-range navigation. They used some structures on their way as navigational landmarks to negotiate. Individual hornets are supposed to have their own landmarks. Olfactory cues seem to be used to find a new feeding place or to recruit other member. In the approach flight hornets seemed to use multiple visual cues such as the visual characteristics of the feeder and the wider scenery around the feeder. Even if the feeder in training was removed during the test, they flew with a smooth course as if they were pin-pointing the missing feeder, but without sitting on the ground. Hornets learnt how to fly to reach the feeder without external cues after passing by the last visual landmark under conditions with extremely poor visual cues.

The present work suggests that hornets retain multiple navigational cues during repeated foraging behavior, and which cues they use seems to depend upon environmental conditions.

Hypodermal injection of Toughmac-E, a digestive mixture composed of nine digestive components, or Molsin induced perturbation of the wing color pattern in 0- to 2-day-old pupae of Papilio xuthus, but had no effect on prepupae or 3- to 4-day-old pupae. The effective component in Toughmac-E was identified as Molsin, an acid carboxypeptidase of Aspergillus saitoi which specifically liberates tyrosine and phenylalanine from the C-terminal residues of proteins. The pattern perturbation occurred in either side of the fore- and hindwings of both sexes. When this enzyme was administered, stronger melanization than in the normal wings was found in the whole wings of most butterflies, but in other butterflies, the yellow region was enlarged. These findings suggest that the pattern perturbation was caused by changes in the levels of melanin and papiliochromes in scales. Melanin is a black pigment and papiliochromes are yellow pigments; their common precursor is dopamine. The normal pattern is formed by a predetermined balance of melanin and papiliochromes, whereas the deposit of an excess amount of tyrosine and/or phenylalanine disturbs this balance and results in perturbation of the color pattern. Administration of L-dopa or dopamine had no effect on the wing pattern. When the activity of an endogenous acid carboxypeptidase similar to Molsin appears in the early pupa, the summed activities of the endogenous and exogenous acid carboxypeptidases must induce a pattern perturbation. The relations between the endogenous acid carboxypeptidase and its probable substrate, the reserve protein, and the physiological roles of these relations in the regulation, utilization and excretion of amino acids are discussed.

To establish techniques for chimera formation and to obtain further knowledge of chimerism, chimeric loach were produced using the wild strain as the donor and the orange strain as the recipient by cell transplantation. Transplantation between embryos at two different stages was performed to achieve efficient chimera formation. In the combination of the early-mid-blastula as the donor and the late-blastula as the recipient, 100–150 blastomeres were injected into the blastoderm of the recipient and the rate of chimera formation was 46.2%. On the other hand, in the combination of early-mid-blastula and early-gastrula, only 30 blastomeres were injected and the rate of chimera formation was 80.0%. These results demonstrating the combination of embryonic stages may provide a key for efficient chimera formation. We also compared the number of melanophores on chimeric larvae with that on donor cells labelled with latex beads; it was found that the number of transplanted cells has a profound effect on chimerism, whereas the site of pigmentation is not always in agreement with the site of actual transplantation of donor cells.

A number of avian species are difficult to sex morphologically, especially as nestlings. Like other avian species, many species of Falconiformes are sexually monomorphic. Therefore, it is desirable that new methods based on DNA analysis are established in Falconiformes and other sexual monomorphic species. We identified sex in Falconiformes by two alternative methods. First, we used a sexing method based on the intronic length variation between CHD1W and CHD1Z using primers flanking the intron. In this method, two species of Falconidae could be identified for sexing. However, six species of Accipitridae could not, because they have few length variations. The second method used was based on differences in sequences between CHD1W and CHD1Z. From sequence analysis, a 3′-terminal mismatch primer on point mutation conserved among Falconiformes was designed, and identification of sex with the amplification refractory mutation system (ARMS) was performed. This method could identify sex in all species tested. In addition, because the 3′-terminal mismatch primer was designed on a point mutation conserved among Falconiformes, ARMS with these primers may identify sex in all Falconiformes. These are simple and rapid sexing methods, since only polymerase chain reaction (PCR) and agarose electrophoresis are required. In conclusion, sex identification by an alternative PCR approach based on intronic length variation and on differences in sequences between CHD1W and CHD1Z proved applicable to and useful for Falconiformes.

Most urodeles undergo internal fertilization and sperm are directly inseminated onto the surface of egg-jelly. Feature of sperm motility induced on the egg-jelly was examined in the newt, Cynops pyrrhogaster. When sperm were directly inseminated onto an egg-jelly, sperm motility was immediately induced on its surface. The egg-jelly of C. pyrrhogaster was composed of six sublayers that were added by turns in oviduct. When the eggs with various sets of the sublayers were obtained and sperm were inseminated onto the egg-jelly, the immediate activity for the initiation of sperm motility was observed only on the outermost sublayer. Similarly, the immediate initiation of sperm motility was induced in the sperm suspended in the extract of the egg-jelly (JE). The initiation of sperm motility was affected by the external pH, and the motility was activated in the moving sperm. A K -channel antagonist, charybdotoxin (CTX), or a Ca² -channel antagonist, gallopamil inhibited the initiation of sperm motility in a dose dependent manner. These results demonstrated the feature of the mechanism regulating sperm motility under stable surroundings in the internal fertilization of amphibians.

In stonefish, changes in plasma total Ca and calcitonin levels were examined after administration of a high-Ca solution into the stomach. Blood was taken successively at 0, 1, 3, 9, 33, and 81 hr from a fine tube cannulated into the aortic bulb. Plasma Ca levels increased acutely at 1 hr and attained the peak after 3–9 hr of the administration. Although plasma calcitonin levels did not exhibit conspicuous changes for 1–3 hr, they began to rise significantly at 33 hr. The plasma Ca level began to decline significantly at 33 hr, although the level was still significantly higher than the initial level. At 33 hr, however, the plasma calcitonin level still continued to increase. At 81 hr, the plasma Ca level had returned to the initial level. At that time, the plasma calcitonin level was also significantly lower than that at 33 hr. These results suggest that, in stonefish, the ultimobranchial gland has the ability to respond physiologically to rises in plasma Ca levels, to secrete calcitonin, and to cease the secretion when the plasma Ca levels return to the initial level.

A new species of polyclad flatworm from Papua New Guinea is described. It is found symbiotic in the ophiuroid Ophiothrix purpurea von Martens, 1867 (Echinodermata: Ophiuroidea). Apparently it belongs to the taxon Discoplana Bock, 1913 and can be distinguished from the six previously described Discoplana species by its very short ejaculatory duct and a penial papilla covered with a penial sheath, but without any true sclerotised structures such as a stylet or spines. The cladistic analysis of the Discoplana/Euplana species, based on morphological features and including two outgroups, reveals that all species of Discoplana, except D. pacificola, form a monophyletic taxon, that is not a synonym of Euplana Girard, 1893. Therefore the name Discoplana is conserved and the new species will be described as Discoplana malagasensis sp. nov. A key for the Discoplana/Euplana group is provided. In this key the bio-geographical distribution and possible synonyms are given.

A new species of Alcyonosyllis Glasby and Watson, 2001 (Polychaeta: Syllidae: Syllinae) is described from shallow water of Shimoda, Izu Peninsula, Japan. The worms were commensal with the gorgonian Melithae flabellifera (Kükenthal, 1909). Alcyonosyllis glasbyi n. sp. differs from all other species of the genus in having a single chaeta per parapodium. The genus is new to Japan.

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°C from before hatching died within one day after hatching. By contrast, juveniles kept outdoors (7.0–20.9°C) could develop at least until second instar larvae. Temperature >23°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.