Zoological Science

Foragers of the Japanese honeybee (Apis cerana japonica) were attracted by flowers of an oriental orchid (Cymbidium floribundum) and were observed to carry the pollinia on their scutella. After the removal of pollinia from the flowers, their labial color changed from white to reddish brown. Both artificial removal of pollinia and ethrel treatment of the flowers also induced this labial color change. Labia in color-changed flowers showed a decreased reflectance of wavelengths less than 670 nm compared to control intact flower. Both reflectance irradiance spectra and ultraviolet photographs showed that only the nectar guide in white (unchanged) flowers reflected ultraviolet light, and that this reflectance decreased with labial color change. Dual choice experiments showed that the honeybee foragers preferentially visited flowers having white labia rather than reddish brown. We suggest that Japanese honeybees discriminate between the floral phases of C. floribundum using color vision.

We analyzed partial sequences (approx. 630 bp) of the 18S nuclear ribosomal RNA gene (rDNA) in the Japanese Eothenomys voles, E. andersoni and E. smithii, to evaluate their evolutionary complexity. There were two rDNA types of sequences with a few substitutions and an indel, and both species alternatively carried one of the two rDNA types, irrespective of their classifications. One of the rDNA types was mainly distributed in Eastern Japan, and the other in Western Japan. Such rDNA type distributions were not related to the species classifications. The complex genomic elements across both species were confirmed by previous studies of nuclear, mitochondrial, and Y chromosomal genes, and our current data agree with previous findings. Previous studies indicated that the genome constitutions of the Japanese Eothenomys species may have been caused by interspecific genetic exchanges at an early period after the speciation into E. andersoni and E. smithii from their common ancestor, as neither species crosses at present. In addition, the present results also suggest the additional hypothesis that their common ancestor might have two (or more) rDNA gene types, and that one of the rDNA types was fixed in each population of both species during their colonization processes.

The physical habitat characteristics associated with spatial distribution patterns of the freshwater mussel Pronodularia japanensis, which is used for oviposition by the Tokyo bitterling Tanakia tanago, were investigated in a small stream within a Tokyo bitterling protected area. The distribution of the mussels was found to be in an under-dispersed, non-random spatial pattern. Mussel occurrence correlated negatively with sediment softness, and positively with flow velocity, while mussel abundance was associated negatively with sediment softness and positively with sediment type (particle size). Furthermore, mussels were scarce in riverbed areas with a lack of sediment. These correlations suggest that the population dynamics of mussels and Tokyo bitterling may be influenced by changes in stream sediment conditions. To conserve the symbiosis between Tokyo bitterlings and mussels, a suitable benthic environment is required.

Monitor lizards belong to the largest and the most sexually dimorphic lizards in terms of size, making this group an ideal model for studies analyzing ontogenetic causes of sexual dimorphism. Understanding of these ontogenetic factors is essential to the current discussion concerning patterns of sexual dimorphism in animals. We examined the ontogenetic trajectories of body weight and snout-vent length to analyze the emergence of sexual size dimorphism. Experimental animals were 22 males and 13 females of mangrove-dwelling monitors (Varanus indicus) hatched at the Prague Zoo. They were regularly weighed and measured up to the age of 33–40 months, and subsequently sexed by ultrasonographic imaging. The logistic growth equation was used to describe and analyze the observed growth patterns. Our results confirm considerable sexual size dimorphism in the mangrove monitor. The mean asymptotic body weight of males was nearly three times higher than that of females. As the body size of male and female hatchlings is almost equal, and the growth rate parameter (K) of the logistic growth equation as well as the absolute growth rate up to the age of 12 months do not differ between the sexes, size differences between fully grown males and females should be attributed to timing of the postnatal growth. Males continue to grow several months after they reach the age when the growth of females is already reduced. Therefore, the sexual size dimorphism emerges and sharply increases at this period.

We examined whether a gonadotropin-releasing hormone (GnRH)-like peptide is present in the nerve ganglion of the chiton Acanthopleura japonica (Mollusca, Polyplacophora) using reverse-phase high performance liquid chromatography (rpHPLC) combined with time-resolved fluoroimmunoas-say (TR-FIA) analysis, and immunohistochemistry. An extract of the chiton head region showed a similar retention time to that of synthetic lamprey GnRH-II on rpHPLC combined with TR-FIA analysis using a rabbit polyclonal antibody raised against chicken GnRH-II (aCII6). Cell bodies immunostained with LRH13 (a mouse monoclonal antibody raised against the common amino acid sequence of vertebrate GnRH) were detected in the cerebrobuccal ring (CBR). Cell bodies immunostained with aCII6 were not only observed in the CBR but also in the lateral nerve cord (LCo). Fibers immunostained with LRH13 and aCII6 were widely distributed throughout the central nervous system in the CBR, subradular ganglion (SubRG), pedal nerve cord (PCo), pedal commissure (P/PCom), lateropedal commissure (L/PCom), and from the LCo to the suprarectal commissure (SupRecCom). The cell bodies and fibers immunostained with these two antisera were distinguishable by dual-label immunohistochemistry. These results suggest that multiple GnRH-like peptides are present in the nerve ganglion of the chiton Acanthopleura japonica.

Detailed examination of early development and growth of Cuban gar (Atractosteus tristoechus) was conducted using morphologic and morphometric characters. Larvae were reared at a constant water temperature (28 ± 1°C) from hatching to 18 days after hatching (DAH). Observation of the disappearance, reduction, or appearance of external structures, pigment characteristics, and behavior, identified three developmental stages (attached 0–3 DAH; transitional 4–10 DAH; and free-swimming 11–18 DAH). For the 18 day trial, the average growth rate was 1.30 mm/d and the specific growth rate averaged 10.2%/d. The slowest growth rates (0.02 mm/d and 2.8%/d, 7–11 DAH) coincided with the exhaustion of yolk reserves and the transition from endogenous to exogenous feeding. In addition to the slowest growth rates, the most dramatic morphological changes in A. tristoechus were observed during the transition from endogenous to exogenous feeding. Five total length (TL) groupings were established using 25 morphologic and morphometic characters. The characters snout length, pelvic fin length, snout width at nares, head length, and head width best identified length groupings. As the larvae developed, the snout and head lengthened and narrowed. Proportionally, the snout was narrower and the head longer in Cuban gar larvae than in other lepisosteid larvae.

In order to clarify the morphological differences between two subspecies of the red fox (Vulpes vulpes) on the Japanese Islands and test the validity of Bergmann's rule, we examined geographical variations in 25 cranial and 24 dental characters in V. v. schrencki from Hokkaido and V. v. japonica from the other main islands of Japan (Honshu, Shikoku, and Kyushu). Many skull measurements, including the male greatest length, condylobasal length, and the length of upper and lower tooth rows, were significantly larger for V. v. japonica than for V. v. schrencki, whereas most tooth measurements, especially the length of molars and premolars, in V. v. schrencki were larger than those in V. v. japonica. Although the two subspecies were morphologically well-differentiated from each other, the results did not support that they have evolved following Bergmann's rule of adaptation to cold climates. Based on consideration of the relatively large differences of their tooth sizes, which are not easily influenced by food abundance, and previous genetic research on the different migration histories of the two subspecies, the morphological differences detected in the present study may have resulted not only from the present ecological differences between the two subspecies, but also from the difference of migration history and evolutionary constraints.

The black-throated tit, Aegithalos concinnus, and long-tailed tit, A. caudatus, are two widely-distributed species of Aegithalidae. They are thought to be monomorphic and thus difficult to differentiate between sexes in the field. We determined the sex of 296 black-throated tits and 129 long-tailed tits using DNA analysis, evaluated their sexual size dimorphism, and developed discriminant models to identify sex based on morphometries, including bill length, bill depth, bill-head length, maximum tarsus length, tarsus width, wing length, tail length, total body length, and body mass. Both species were sexually dimorphic in size, with males having larger measurements than females, except for bill length in black-throated tits, and both bill length and body mass in long-tailed tits. Moreover, both species showed similar sexual size dimorphism (SSD) among the morphological features, with tail length having the highest SSD value. The multivariate discriminant models for sex identification had an accuracy of 82% for both species, which was slightly higher than the best univariate discriminatory model for each species. Because of the complicated nature of the multivariate model, we recommend univariate models for sex identification using D = 0.491 × tail length - 24.498 (accuracy 80%) for black-throated tits and D = 0.807 × wing length - 45.934 (accuracy 78%) for long-tailed tits. Females in both species showed generally higher morphological variation than did males, resulting in lower accuracies in all discriminate functions regardless of univariate or multivariate approach. This could be the result of a sex-biased selective pressure in which males have higher selective pressures for these morphological features.

The aim of the present work is to conduct a comparative study of oxidative states in the nymphs of four species of Plecoptera belonging to the superfamily Perloidea: Perla marginata (Panzer, 1799) (family Perlidae), Guadalgenus franzi (Aubert, 1963), Isoperla curtate Navás, 1924, and lsoperla grammatica (Poda, 1761) (family Perlodidae) in relation to their ecological and biological characteristics. For this, the activity of the following antioxidant enzymes was determined: superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPX), glutathione transferase (GST), and DT-diaphorase (DTD), together with lipid peroxidation. Glucose 6-phosphate dehydrogenase (G6PDH) was also determined. The four species studied were selected based on significant ecological and biological differences. The results obtained when studying different indicative parameters of the oxidative state of the nymph of different species showed that each has an important enzymatic antioxidant potential, and that differences among species are conditioned by the duration of the nymphal development period more than by whether they come from permanent or temporary habitats. Thus, Plecoptera, although traditionally considered as typical inhabitants of permanent waters, seem to have sufficient variability in physiological mechanisms, together with behavioral and ecological adaptations, to cope with potentially unfavorable conditions that may occur in temporary waters.

Didemnum molle, a colonial ascidian that harbors the symbiotic cyanophyte Prochloron spp., is distributed throughout the coral reefs of the Indo—West Pacific Ocean. Several morphotypes of D. molle are characterized by the color and size of their colonies. Previous molecular phylogeny inferred from gene sequences for the cytochrome oxidase subunit I (COI) identified four morphotypes (i.e., gray, brown, white, and large) from several sites in the Ryukyu Islands, Japan. With the addition of 17 specimens, including another morphotype (small), from several collection sites (Taiwan and the Ryukyus), the present report demonstrates genetic separation among the five morphotypes based on COI sequences. A number of sexually mature specimens of the different morphotypes were collected at the same times and sites, indicating reproductive isolation among morphotypes.

Temnocephala brevicornis Monticelli, 1889 is the only species of the genus Temnocephala Blanchard, 1849 reported from chelonians to date. During a survey of the species of Temnocephala extant in southern Uruguay, two new species were found on the chelonian Hydromedusa tectifera Cope, 1869. They are described here as Temnocephala pereirai n. sp. and Temnocephala cuocoloi n. sp. Both resemble T. brevicornis, but differ in the morphometry of the penial stylet, and in qualitative details of the reproductive complex. Temnocephala pereirai n. sp. differs from T. brevicornis by having a massive, cylindrical sphincter in the distal portion of the vagina, and a seminal vesicle that opens into the subpolar to equatorial portion of the contractile vesicle. In addition, the penial stylet in Temnocephala pereirai n. sp. is large in relation to body size, straight and more slender, having the distal portion of its shaft slightly sinuous, and a smaller introvert equipped with about 16 distal crowns of smaller spines. Temnocephala cuocoloi n. sp. is most similar to T. brevicornis, but differs by having a smaller, curved penial stylet that has a smaller introvert in relation to stylet size, with about 10 distal crowns of smaller spines. A key to the species of the Temnocephala from chelonians is provided. This study supports the validity of the following characters previously proposed for the taxonomy of the genus Temnocephala: the shape of the sphincters in the female reproductive system, the shape of the penial stylet, and the number, size, and position of spines in the introvert.