Zoological Science

Evolution of avian life histories is typically strongly influenced by both altitude and latitude. To date, most studies have investigated the effects of extreme differences in altitude and latitude on variation in reproductive traits. Studies based on small altitude and latitude spans are needed to better understand the resolution of selective pressures. We compared several aspects of russet sparrow (Passer cinnamomeus) breeding biology between a low-altitude (200 m) and a high-altitude (1,500 m) population in China, representing a relatively small altitudinal gradient (1,300 m). High-altitude birds initiated breeding significantly later compared to their low-altitude counterparts. Interestingly, breeding season was significantly longer in the high-altitude site (57 vs. 84 d). Lowland sparrows laid larger clutches (4.92 vs. 4.09 eggs) and showed greater fledging success (4.20 vs. 3.46 fledgings) than did upland birds. Variation in life history traits thus appears to occur even along a smallscale altitudinal gradient. We suggest that the longer breeding season and smaller clutch size in the highland population may be an adaptation or acclimation to compensate for the reduced annual productivity resulting from unfavourable or sub-optimal habitats for these sparrows.

Sichuan sika deer (Cervus nippon sichuanicus) is an endangered and endemic subspecies of sika deer to Sichuan Province, China. According to our observations in the wild, the Sichuan sika deer makes alarm signals in the presence of actual or potential predators. In order to test the variation of the rhythmic alarm calls in some sex/age classes and different risk contexts, we recorded alarm calls of Sichuan sika deer from 2 October to 30 November 2008 and from 4 April to 5 September 2009 in the Tiebu Nature Reserve, Zoige County, Sichuan Province, China, and made acoustic analysis of these alarm calls. The results showed that the fundamental frequencies of alarm signals of Sichuan sika deer tended to decrease with age, and were significantly higher for females than for males. Duration tended to increase with age, and was significantly longer for males than for females. The fundamental frequencies and duration of alarm calls in adults were significantly higher and shorter respectively in high-risk than in moderate-risk contexts.

The morphologies of male genitalia often appear harmful or aggressive, as if they may inflict physical damage upon females during copulation. Such male genitalia are often thought to function in intra- and intersexual interactions during mating. In the carabid genus Carabus, division Spinulati, males possess a spine (spinula) on the intromittent organ, of which function is unknown. To reveal the function of the spinula, we studied the mating behavior and genital coupling of a Spinulati species, Carabus (Limnocarabus) clathratus. The males positioned the spinula along the inner wall of the vaginal opening throughout copulation. This placement created a small dent and subsequently a melanized patch (wound) on the vaginal wall, but the spinula rarely penetrated the vaginal wall. The spinula did not reach the innermost part of the vagina where the spermatophore is deposited. These results suggest that the spinula is not used for inflicting damage on female genitalia or manipulating spermatophores of rival males. During spermatophore formation, the male partially withdrew the aedeagus, and only the aedeagal tip and endophallus remained within the female. By placing the spinula against the vaginal wall, the male could hold the endophallus within the vaginal chamber in the unstable copulatory posture. Thus, our observations suggest that the spinula primarily functions as an “anchor” to maintain the coupling of the male and female genitalia and thereby ensure insemination.

The morphologies of male genitalia often appear harmful or aggressive, as if they may inflict physical damage upon females during copulation. Such male genitalia are often thought to function in intra- and intersexual interactions during mating. In the carabid genus Carabus, division Spinulati, males possess a spine (spinula) on the intromittent organ, of which function is unknown. To reveal the function of the spinula, we studied the mating behavior and genital coupling of a Spinulati species, Carabus (Limnocarabus) clathratus. The males positioned the spinula along the inner wall of the vaginal opening throughout copulation. This placement created a small dent and subsequently a melanized patch (wound) on the vaginal wall, but the spinula rarely penetrated the vaginal wall. The spinula did not reach the innermost part of the vagina where the spermatophore is deposited. These results suggest that the spinula is not used for inflicting damage on female genitalia or manipulating spermatophores of rival males. During spermatophore formation, the male partially withdrew the aedeagus, and only the aedeagal tip and endophallus remained within the female. By placing the spinula against the vaginal wall, the male could hold the endophallus within the vaginal chamber in the unstable copulatory posture. Thus, our observations suggest that the spinula primarily functions as an “anchor” to maintain the coupling of the male and female genitalia and thereby ensure insemination.

The sika deer (Cervus nippon) once inhabited the entire Tohoku District, the northeastern part of the main island of Japan. Currently, they are isolated as three discontinuous populations on Mt. Goyo, the Oshika Peninsula, and Kinkazan Island. To assess the genetic diversity and relationships among the sika deer populations in the Tohoku District, we analyzed the mitochondrial DNA D-loop sequences from 177 individuals. We detected a total of five haplotypes. Three haplotypes were present in the population from Mt. Goyo at a haplotype diversity of 0.235 ± 0.061, two haplotypes in the population from the Oshika Peninsula at 0.171 ± 0.064, and only one haplotype was detected in the population from the Kinkazan Island. A significant genetic differentiation was observed among all population pairs. Collectively, our data supports the observed population bottlenecks in the past. Four of the five haplotypes were specific to one of the three populations, whereas only one haplotype was shared between the Mt. Goyo and the Oshika Peninsula populations. This common haplotype may indicate a common ancestral population in the Tohoku District. Conversely, the D-loop haplotypes were completely different among the Kinkazan Island and Oshika Peninsula populations. The lack of a shared haplotype indicates that female gene flow between the two populations is very limited and that the 0.6 km strait acts as a strong barrier.

The sika deer (Cervus nippon) once inhabited the entire Tohoku District, the northeastern part of the main island of Japan. Currently, they are isolated as three discontinuous populations on Mt. Goyo, the Oshika Peninsula, and Kinkazan Island. To assess the genetic diversity and relationships among the sika deer populations in the Tohoku District, we analyzed the mitochondrial DNA D-loop sequences from 177 individuals. We detected a total of five haplotypes. Three haplotypes were present in the population from Mt. Goyo at a haplotype diversity of 0.235 ± 0.061, two haplotypes in the population from the Oshika Peninsula at 0.171 ± 0.064, and only one haplotype was detected in the population from the Kinkazan Island. A significant genetic differentiation was observed among all population pairs. Collectively, our data supports the observed population bottlenecks in the past. Four of the five haplotypes were specific to one of the three populations, whereas only one haplotype was shared between the Mt. Goyo and the Oshika Peninsula populations. This common haplotype may indicate a common ancestral population in the Tohoku District. Conversely, the D-loop haplotypes were completely different among the Kinkazan Island and Oshika Peninsula populations. The lack of a shared haplotype indicates that female gene flow between the two populations is very limited and that the 0.6 km strait acts as a strong barrier.

The microvascularization of the collared peccary (Tayassu tajacu) placenta was studied by vascular casts and immunolocalization of α-smooth muscle actin and vimentin, to identify the threedimensional organization and vascular flow interrelation in the microvasculature between the maternal and fetal compartments of the placentae. The immunolocalization of vimentin in the vascular endothelium and in the smooth muscle cells of blood vessels showed indented capillaries along the uterine epithelium and the trophoblast at the sides of complementary maternal and fetal microfolds, or rugae. This confers the three-dimensional structure observed in vascular casts. On the maternal side, casts demonstrated uterine folds coated by with primary and secondary ridges, and by areolae dispersed between these ridges. The arteriole runs through the center/middle of ridges, branching at the top into a microvascular network wall in a basket-like fashion. At the base of these baskets venules were formed. On the fetal side, arterioles branched centrally in the fetal rugae into a capillary network in a bulbous form, complementary to the opposite maternal depressions forming the baskets. At the base of the bulbous protrusions, the fetal venules arise. The blood vessel orientation in the materno-fetal interface of the placentae of collared peccaries suggests a blood flow pattern of the type countercurrent to crosscurrent. The same pattern has been reported in domestic swine demonstrating that, even after 38 million years, the Tayassuidae and Suidae families exhibit similar placental morphology, which is here characterized at the microvascular level.

The microvascularization of the collared peccary (Tayassu tajacu) placenta was studied by vascular casts and immunolocalization of α-smooth muscle actin and vimentin, to identify the threedimensional organization and vascular flow interrelation in the microvasculature between the maternal and fetal compartments of the placentae. The immunolocalization of vimentin in the vascular endothelium and in the smooth muscle cells of blood vessels showed indented capillaries along the uterine epithelium and the trophoblast at the sides of complementary maternal and fetal microfolds, or rugae. This confers the three-dimensional structure observed in vascular casts. On the maternal side, casts demonstrated uterine folds coated by with primary and secondary ridges, and by areolae dispersed between these ridges. The arteriole runs through the center/middle of ridges, branching at the top into a microvascular network wall in a basket-like fashion. At the base of these baskets venules were formed. On the fetal side, arterioles branched centrally in the fetal rugae into a capillary network in a bulbous form, complementary to the opposite maternal depressions forming the baskets. At the base of the bulbous protrusions, the fetal venules arise. The blood vessel orientation in the materno-fetal interface of the placentae of collared peccaries suggests a blood flow pattern of the type countercurrent to crosscurrent. The same pattern has been reported in domestic swine demonstrating that, even after 38 million years, the Tayassuidae and Suidae families exhibit similar placental morphology, which is here characterized at the microvascular level.

A population of the Australian mudskipper, Periophthalmus minutus, was found to inhabit mudflat that remained uncovered by tide for more than 20 days in some neap tides. During these prolonged emersion periods, P. minutus retreated into burrows containing little water, with a highest recorded salinity of 84 ± 7.4 psu (practical salinity unit). To explore the mechanical basis for this salinity tolerance in P. minutus, we determined the densities of mitochondria-rich cells (MRCs) in the inner and outer opercula and the pectoral fin skin, in comparison with P. novaeguineaensis, from an adjacent lower intertidal habitat, and studied morphological responses of MRCs to exposure to freshwater (FW), and 100% (34–35 psu) and 200% seawater (SW). Periophthalmus minutus showed a higher density of MRCs in the inner operculum (3365 ± 821 cells mm^-2) than in the pectoral fin skin (1428 ± 161) or the outer operculum (1100 ± 986), all of which were higher than the MRC densities in p. novaeguineaensis. No mortality occurred in 100% or 200% SW, but half of the fish died within four days in FW. Neither 200% SW nor FW exposure affected MRC density. Transfer to 200% SW doubled MRC size after 9–14 days with no change in the proportion of MRCs with apical pits or plasma sodium concentration. In contrast, transfer to FW resulted in a rapid closing of pits and a significant reduction in plasma sodium concentration. These results suggest that P. minutus has evolved morphological and physiological mechanisms to withstand hypersaline conditions that they may encounter in their habitat.

A population of the Australian mudskipper, Periophthalmus minutus, was found to inhabit mudflat that remained uncovered by tide for more than 20 days in some neap tides. During these prolonged emersion periods, P. minutus retreated into burrows containing little water, with a highest recorded salinity of 84 ± 7.4 psu (practical salinity unit). To explore the mechanical basis for this salinity tolerance in P. minutus, we determined the densities of mitochondria-rich cells (MRCs) in the inner and outer opercula and the pectoral fin skin, in comparison with P. novaeguineaensis, from an adjacent lower intertidal habitat, and studied morphological responses of MRCs to exposure to freshwater (FW), and 100% (34–35 psu) and 200% seawater (SW). Periophthalmus minutus showed a higher density of MRCs in the inner operculum (3365 ± 821 cells mm^-2) than in the pectoral fin skin (1428 ± 161) or the outer operculum (1100 ± 986), all of which were higher than the MRC densities in p. novaeguineaensis. No mortality occurred in 100% or 200% SW, but half of the fish died within four days in FW. Neither 200% SW nor FW exposure affected MRC density. Transfer to 200% SW doubled MRC size after 9–14 days with no change in the proportion of MRCs with apical pits or plasma sodium concentration. In contrast, transfer to FW resulted in a rapid closing of pits and a significant reduction in plasma sodium concentration. These results suggest that P. minutus has evolved morphological and physiological mechanisms to withstand hypersaline conditions that they may encounter in their habitat.

The hagfish liver was histochemically examined with special attention to biliary structures around the portal veins. Hepatocytes were organized into tubular structures surrounded by sinusoids. Biliary ductule structures, which resemble the ductal plates transiently appearing in mammalian liver development, were observed around the portal veins, but they did not appear around central veins. Thus, the hagfish liver demonstrates the same basic structure as the mammalian liver; that is, a vascular system from portal to central veins via sinusoids, and portal triad structures consisting of portal veins, hepatic arteries, and intrahepatic bile ducts. The epithelial cells of the ductal platelike structures strongly expressed cytokeratin, had some lectin-binding sites, and were delineated by the basal lamina, which was reactive for periodic acid-Schiff (PAS) staining and Iectin histochemistry. The lumina of the ductal plate-like structures were comparatively small and heterogeneous in diameter around the portal veins, suggesting that the biliary structures may not be efficient for bile secretion. The epithelial cells of the gall bladder had a simple columnar shape and were a PAS-positive cytoplasm. Those of bile ducts near the hilus, including extrahepatic and hepatic ducts, were simple columnar or cuboidal cells, and had large lumina. The cytoplasm in these cells was PAS-positive. These phenotypes with the expression of lectin-binding sites were clearly different from those of the ductal plate-like structures in the liver proper, suggesting that the extrahepatic and intrahepatic biliary structures may have different developmental origins.

Based on LM, TEM, and histochemical methods, the study describes the specific structure of subepidemal capillary loops in the integument of the hippopotamus (Hippopotamus amphibius). At 25– 60 µm, the diameter of the capillaries was more than twenty times larger than those found in other mammals, as was the diameter of the epidermal contact area of the hairpin turn, which had enlarged up to 200–400 µm². At about 13,400, the number of loops per cm² was three times higher than in the few other mammalian species measured to date. The remarkable sheath (thickness 2– 20 µm) of the capillary loops consists of a multitude of fine collagen IV fibres, which were in direct contact with the epidermal stratum (str.) basale, emphasizing an origin from the lamina fibroreticularis of the basement membrane. Additionally, the sheath contained many regions filled with free fatty acids. All observations confirmed the view that the walls of the subepidermal capillaries in the hippopotamus are adapted to withstand high blood pressure, permitting a high rate of blood vesselbased heat transfer from the periphery of the body. Until now this function is only known as an important thermoregulatory response in highly active mammals, e.g. dolphins. However, under hot climatic conditions but without strong exercise for cooling, such ability could be an effective and energy-saving procedure in semi-aquatic mammals.

Based on LM, TEM, and histochemical methods, the study describes the specific structure of subepidemal capillary loops in the integument of the hippopotamus (Hippopotamus amphibius). At 25– 60 µm, the diameter of the capillaries was more than twenty times larger than those found in other mammals, as was the diameter of the epidermal contact area of the hairpin turn, which had enlarged up to 200–400 µm². At about 13,400, the number of loops per cm² was three times higher than in the few other mammalian species measured to date. The remarkable sheath (thickness 2– 20 µm) of the capillary loops consists of a multitude of fine collagen IV fibres, which were in direct contact with the epidermal stratum (str.) basale, emphasizing an origin from the lamina fibroreticularis of the basement membrane. Additionally, the sheath contained many regions filled with free fatty acids. All observations confirmed the view that the walls of the subepidermal capillaries in the hippopotamus are adapted to withstand high blood pressure, permitting a high rate of blood vesselbased heat transfer from the periphery of the body. Until now this function is only known as an important thermoregulatory response in highly active mammals, e.g. dolphins. However, under hot climatic conditions but without strong exercise for cooling, such ability could be an effective and energy-saving procedure in semi-aquatic mammals.

Libytheinae is a morphologically unusual group of butterflies whose external anatomy has never been described in detail. The subfamily is considered basal within the diverse family Nymphalidae, and libytheines retain many plesiomorphic features, such as a functional female foreleg and smooth larva. In this paper, we describe and illustrate the adult morphology of Libythea celtis Laicharting, make comparisons to other nymphalid subfamilies, and homologize terms whenever possible. We describe major sclerites, sulci, and the presence of cranial and lateral pits on the tibia, the latter which appear to be previously unreported from the lepidopteran literature. When present, sexual dimorphism and geographic variation are noted. It is hoped that the present paper will help stabilize inconsistencies in morphological terms in nymphalid butterflies.