Suckling behaviors are useful to better understand mother–offspring relationships. However, in many species, knowledge about nighttime suckling behavior is sparse. In the present study, we investigated suckling behavior in four calves of the reticulated giraffe (Giraffa camelopardalis reticulata) and their mother in the Kyoto City Zoo, Japan, by analyzing video clips of 9614 h. The relation between a calf's age and the mean duration and frequency of suckling were consistent with the results of previous daytime studies: the longest duration (80 ± 4 s) and highest frequency (0.41/h) of suckling occurred when the calves were young, and decreased thereafter. Suckling frequency was lower at night than during the day, probably because the mother spent much of the time resting at night. The mother initiated suckling behavior in offspring until six months of age. Most often, the mother terminated suckling except during the first few weeks after birth. Additionally, we reported that the average weaning period was 448 days (n = 5) and that the last born (sixth) continued to suckle for more than 750 days, which is a rare phenomenon. We believe the present report helps to fill the lack of knowledge about nighttime suckling behavior in giraffe.

The outer dynein arm-docking complex (ODA-DC), which was first identified in the green alga Chlamydomonas reinhardtii, is a protein complex that mediates the binding of axonemal dynein and doublet microtubules. To gain a better understanding of the evolutionary conservation and functional diversity of the ODA-DC, we knocked down a homolog of DC2, a major subunit of the ODA-DC, in the planarian Schmidtea mediterranea. Planaria are carnivorous flatworms that move by beating cilia on their ventral surface against a secreted mucus layer. These organisms have recently been used for cilia research because knockdown of flatworm genes by RNA interference (RNAi) is readily achieved through feeding with double-stranded RNA (dsRNA). Lack of DC2 in S. mediterranea caused several defects in cilia, including low beat frequency, decreased ciliary density, and a reduction in ciliary length. The loss of DC2 function C. reinhardtii mutant oda1 shows slow jerky swimming, but has two flagella of almost normal length. These data suggest that the function of a DC2 homolog in S. mediterranea cilia may be somewhat different from DC2 in C. reinhardtii flagella.

Olfaction plays an important role in a fish's life. Its value may differ at different developmental stages, depending on the feeding style of the species. The goal of the present study was to investigate the olfactory organ of a species that feeds mainly on algae— the bushymouth catfish, Ancistrus dolichopterus—at developmental stages from olfactory placode to the definitive olfactory chamber. For this study, we used light and electron (scanning) microscopy. The topography of the olfactory placode of A. dolichopterus is typical for teleostei. Formation of olfactory pit takes place at the same time as rostral elevation formation. Rostral elevation participates in the formation of the nasal bridge and anterior tubular nostril. It was found out that the anlage of olfactory rosette in A. dolichopterus arises earlier than in most teleostei. However, the number of lamellae does not increase until switching to exogenous feeding. We suppose that the early development of olfactory organ is necessary for intraspecific communication, not just for finding food.

Deep-sea demersal fishes of the Bothrocara hollandi species complex are distributed in the Japan Sea, the Okhotsk Sea, and the northwestern Pacific Ocean. Based on the nucleotide sequences of the nuclear Internal Transcribed Spacer 1 (ITS1) region and the microsatellite analysis, cryptic speciation resulting in the existence of two species (sp. 1 and sp. 2) in the Japan Sea was indicated for this species complex. In the Japan Sea off the San-in district, the westernmost part of the Japanese mainland, the frequency of sp. 2 individuals was highest at a depth of ∼400 m and decreased at both greater and lesser depths. Complete genetic deviation was observed between the individuals of the Japan Sea and the other sea areas, with the exception of a single sp. 2 individual, which shared an ITS1 sequence with an individual from the Pacific Ocean. Furthermore, a microsatellite analysis showed that the individuals of the other sea areas were more closely related to sp. 2 individuals. Two species were thought to have deviated from each other after their isolation from the individuals of the sea areas outside of the Japan Sea, through the occurrence of habitat fragmentation and bottleneck events in the Japan Sea during the glacial periods. Group A, one of two mitochondrial haplotype groups that were reported for the Japan Sea individuals in the previous studies, may have evolved within the lineages of sp. 2.

Immune defense is costly to maintain and deploy, and the optimal investment into immune defense depends on risk of infection. Altitude is a natural environmental factor that is predicted to affect parasite abundance, with lower parasite abundance predicted at higher altitudes due to stronger environmental stressors, which reduce parasite transmission. Using high and low altitude populations of the Turkish blind mole-rat (TBMR) Nannospalax xanthodon, we tested for effects of altitude on constitutive innate immune defense. Field studies were performed with 32 wild animals in 2017 and 2018 from two low- and one high-altitude localities in the Central Taurus Mountains, at respective altitudes of 1010 m, 1115 m, and 2900 m above sea level. We first compared innate standing immune defense as measured by the bacteria-killing ability of blood serum. We then measured corticosterone stress hormone levels, as stressful conditions may affect immune response. Finally, we compared prevalence and intensity of gastrointestinal parasites of field-captured TBMR. We found that the bacteria-killing ability of serum is greater in the mole-rat samples from high altitude. There was no significant difference in stress (corticosterone) levels between altitude categories. Coccidian prevalence and abundance were significantly higher in 2017 than 2018 samples, but there was no significant difference in prevalence, abundance, or intensity between altitudes, or between sexes. Small sample sizes may have reduced power to detect true differences; nevertheless, this study provides support that greater standing innate immunity in high altitude animals may reflect greater investment into constitutive defense.

For seasonal adaptation, the brown-winged green bug Plautia stali (Hemiptera: Pentatomidae) enters reproductive diapause by suppressing juvenile hormone biosynthesis. Plautia stali myoinhibitory peptides (Plast-MIPs) are known to have allatostatic effects and to suppress juvenile hormone biosynthesis. We examined Plast-MIP-producing neurons in the brain with immunohistochemistry and Fourier transform ion cyclotron resonance mass spectrometry. Rabbit polyclonal antiserum against Plast-MIP revealed immunoreactive cells in seven regions of the brain, including the posterior antennal lobe, basal optic lobe, dorsal anterior protocerebrum, ventrolateral protocerebrum, pars intercerebralis, posterior protocerebrum, and dorsal posterior region to the calyx of the mushroom body, aside from the gnathal ganglion. Anatomical locations of the immunoreactive cells in the pars intercerebralis and dorsal posterior region to the mushroom body calyx partly overlapped with the cell body location stained by retrograde dye fills from the corpus allatum and corpus cardiacum complex. Direct mass spectrometry revealed the molecular ion peaks corresponding to the predictive mass of Plast-MIPs in the pars intercerebralis and the corpus allatum–corpus cardiacum complex. Plast-MIP immunoreactivity in different cell types suggests that Plast-MIPs have different functions in the cephalic ganglia. Considering the anatomical location of neurons projecting to the corpus allatum–corpus cardiacum and results of mass spectrometry, Plast-MIP immunoreactive cells in the pars intercerebralis may play a role in suppressing juvenile hormone biosynthesis.

The labial palps of bivalves are thought to be involved in suspension feeding. However, the function of their muscular movements and neural regulation are still unclear. In semi-intact preparations of Mytilus, in which one valve was removed, suspended particles were removed from the labial palps following two kinds of compound movements: torsional and rotational. Both of these compound movements are therefore thought to function in rejection during feeding. These movements were observed in reduced preparations of isolated labial palps with intact cerebral ganglia, and were maintained even after removal of the cerebral ganglia, suggesting that they are generated by the peripheral neural network. Stimulation of the anterior pallial nerve elicited tetanic contraction of the labial palp, followed by secondary responses, including torsional movement. Secondary responses were dramatically reduced by a high concentration of divalent cations, in which polysynaptic pathways were inhibited. Hence, the cerebral ganglia may play an excitatory role within the peripheral neural network and the labial palp musculature via the anterior pallial nerve. Administration of serotonin induced repetitive muscular movements, whereas dopamine did not induce muscular movements. Serotonin-induced muscular movements were not elicited under a high concentration of divalent cation condition. In histochemical experiments, both the serotonergic and dopaminergic neural processes and cell body-like structures were widely observed inside the labial palp, the anterior pallial nerve, and the cerebral ganglia. Serotonin may thus contribute to activation of polysynaptic peripheral pathways, which are involved in regulating compound movements.

Iron is an essential element for hemoglobin synthesis during erythropoiesis. Iron overload, in contrast, adversely affects erythropoiesis and causes organ dysfunction. Research using various animal models may help to elucidate pathophysiological mechanisms induced by excess iron. In the present study, we evaluated the relationship between iron metabolism and erythropoietic activity in the African clawed frog, Xenopus laevis. In X. laevis, both erythropoiesis and iron metabolism occur in the liver. First, we developed a method to quantify iron levels in the liver and plasma using 2-nitroso-5-[N-n-propyl-N-(3-sulfopropyl) amino] phenol (Nitroso-PSAP). We then measured iron levels and analyzed hematopoietic parameters in frogs that were orally administered sodium ferrous citrate (SFC). The hepatic iron level increased in the SFC group, but the number of erythrocytes, hematocrit, and hemoglobin concentration did not change, suggesting that the regulation of the production and release of mature erythrocytes in the liver was not directly affected by dietary iron. At four days after administration of 2 mg/kg SFC, the number of immature erythrocytes decreased in the liver. Interestingly, atypical blood cells with hyper-segmented nuclei were observed, identified by acridine orange cell staining; these atypical blood cells were hardly detectable under the steady state. Compared with previously reported results in mice, the increase in the hepatic iron levels was small, but our results indicate that SFC affects hematopoietic activity. These results establish a novel model for iron metabolism and provide new insights into the relationship between iron metabolism and erythropoiesis in vertebrates.

The taxonomic account of the tadpole-parasitic nematode Gyrinicola japonicaYamaguti, 1938, which is the type species of the genus, was reassessed based on syntypes and newly-collected specimens from the type locality. Our redescription of G. japonica addresses the erroneous original description of a spicule in this nematode, and emends the diagnosis of the species. Additionally, molecular phylogenetic trees based on nuclear 18S and 28S rDNA sequences revealed that G. japonica forms a distinctive lineage within the suborder Oxyurina, and this tadpole-specialist is phylogenetically close to the lizard-parasitic nematodes that belong to the family Pharyngodonidae. The results of morphological examination with the aid of molecular phylogenetic trees highlight the systematic uniqueness of this tadpole-parasitic group within Oxyurina, and Gyrinicolidae is accordingly resurrected as a distinctive oxyurinan family, with redefinition of the family and the genus Gyrinicola.

We describe here a new pterobranch, Cephalodiscus planitectus sp. nov. This pterobranch was collected from rocky slopes, at 100–300 m depth, off Jogashima Island, Sagami Bay, Japan. The tubaria of this new species have unique morphological features that differentiate it from known species. The tubaria are usually isolated from one another and have a completely flat and smooth surface that is devoid of erect features and projecting spines. Each has a simple, non-branched tubular cavity that is usually inhabited by a mature animal and its asexually budding offspring. The zooids have three pairs of tentaculated arms. A single bud is produced on the dorsal side of the stalk in adult zooids. In one instance, a live embryo was observed rotating at the bottom of a tubarium. Molecular phylogenetic analysis showed that C. planitectus is a sister group to all other Cephalodiscus species analyzed to date.

Two lineages of stream toads in the genus Ansonia from Malaysian Borneo have long been suspected to be specifically distinct on the basis of molecular data. We assessed the taxonomic status of these lineages using morphological and additional genetic data. In mtDNA phylogeny, each lineage—one from Bario, Kelabit Highlands of Sarawak, the other from Mt. Mulu of Sarawak and the Crocker Range of Sabah—is separated from other congeners by large genetic distances, comparable with those observed between heterospecific species in the genus. These lineages are also morphologically distinguishable from other species, and are considered to represent valid, independently evolving species. We therefore describe them as A. kelabitensis sp. nov. and A. kanak sp. nov.