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Wild Japanese macaques (Macaca fuscata Blyth) living in the highland and lowland areas of Yakushima are known to have different diets, with highland individuals consuming more leaves. We aim to clarify whether and how these differences in diet are also reflected by gut microbial composition and fermentation ability. Therefore, we conduct an in vitro fermentation assay using fresh feces from macaques as inoculum and dry leaf powder of Eurya japonica Thunb. as a substrate. Fermentation activity was higher for feces collected in the highland, as evidenced by higher gas and butyric acid production and lower pH. Genetic analysis indicated separation of highland and lowland in terms of both community structure and function of the gut microbiota. Comparison of feces and suspension after fermentation indicated that the community structure changed during fermentation, and the change was larger for lowland samples. Analysis of the 16S rRNA V3-V4 barcoding region of the gut microbiota showed that community structure was clearly clustered between the two areas. Furthermore, metagenomic analysis indicated separation by gene and pathway abundance patterns. Two pathways (glycogen biosynthesis I and D-galacturonate degradation I) were enriched in lowland samples, possibly related to the fruit-eating lifestyle in the lowland. Overall, we demonstrated that the more leaf-eating highland Japanese macaques harbor gut microbiota with higher leaf fermentation ability compared with the more fruit-eating lowland ones. Broad, non-specific taxonomic and functional gut microbiome differences suggest that this pattern may be driven by a complex interplay between many taxa and pathways rather than single functional traits.

A spontaneous mutation, en

Background Changes in the gut microbial composition is an important response to cope with the seasonal fluctuations in the environment such as food availability. We examined the bacterial gut microbiome of the wild nonhuman primate, Japanese macaque (Macaca fuscata) in Yakushima over 13 months by noninvasive continuous sampling from three identified adult females. Results Dietary composition varied considerably over the study period and displayed marked shifts with the seasons. Feeding of leaves, fruits, and invertebrates were their main foods for at least one month. Diet had a significant influence on the gut microbiome. We also confirmed significant effect of host uniqueness in the gut microbiome among the three macaques. Leaf-dominated diet shaped unique gut microbiome structures where the macaques had the highest alpha diversity and their gut microbiome was enriched with Spirochaetes and Tenericutes. Diet-related differences in the putative function were detected, such as a differentially abundant urea cycle during the leaf-feeding season. Conclusion Both diet and host individuality exerted similar amounts of effect on gut microbe community composition. Major bacterial taxa showed a similar response to monthly fluctuations of fruit and invertebrate feeding, which was largely opposite to that of leaf feeding. The main constituents of fruits and invertebrates are both digestible with the enzyme of the host animals, but that of leaves is not available as an energy source without the aid of the fermentation of the gut microbiome.

Estrogen (E) and progesterone (P) are the major female hormones and are secreted with changing concentration ratios throughout the menstrual cycle. These hormones have been studied individually regarding their physiological function in the skin, but their concentration ratio (E/P) and its effect on the skin has not yet been investigated. The purpose of this study was to clarify the effect of the E/P ratio on skin barrier function. The menstrual cycle was divided into the menstrual, follicular, ovulation, and luteal phases.The E/P concentration ratios corresponding with each phase were added to a three-dimensional epidermal model or normal human epidermal keratinocytes for 5 days. Gene and protein expression levels of several markers of cell differentiation, including loricrin (LOR) and transglutaminase (TGase), were quantified by real-time PCR and western blotting, respectively. Transepidermal water loss (TEWL) of the three-dimensional epidermal model was measured, and ceramide content was quantified by thin-layer chromatography.Gene expression of the epidermal differentiation markers, LOR and TGase, increased when applying the concentration ratio of E/P associated with the menstrual and luteal phases. The LOR protein level decreased from menstrual to luteal phases, and the TGase protein level increased from menstrual to luteal phases. During the same phases, ceramide NS increased and TEWL decreased.Skin barrier function was improved by culturing cells at specific E/P concentration ratios, which would, therefore, be considered beneficial for female skin. This suggests that dysregulated E/P concentration ratios may be the cause of certain skin problems.

We performed a comprehensive fecal metabolite analysis using LC-MS/MS and LC-QTOF-MS approaches as a preliminary study. Feces of Japanese macaques on Yakushima Island were collected from five monkeys at two separate locations. Using the former methodology, 59 substances such as free amino acids, nucleotides, nucleosides and nucleic acid bases, and organic acids in the citrate cycle were quantitatively detected and successfully differentiated in two different monkey groups by the concentrations of nucleic acid metabolites and free amino acids. In the latter, around 12,000 substances were detected both by positive and negative mode in each sample. Differences in signal intensities were observed between two monkey groups in the concentrations of plant secondary metabolites such as cyanogenic glycosides, flavonoids, and phenolics.

Within the gastrointestinal tract, the physiochemical microenvironments are highly diversified among the different stages of food digestion. Accordingly, gut microbiome composition and function vary at different gut sites. In this study, we examine and compare the compositional and functional potential between the stomach and colonic microbiome of wild Japanese macaques (Macaca fuscata yakui) living in the evergreen forest of Yakushima Island. We find a significantly lower microbial diversity in the stomach than in the colon, possibly due to the stomach's acidic and aerobic environment, which is suboptimal for microbial survival. According to past studies, the microbial taxa enriched in the stomach are aero- and acid-tolerant. By functional prediction through PICRUSt2, we reveal that the stomach microbiome is more enriched in pathways relating to the metabolism of simple sugars. On the contrary, the colonic microbiota is more enriched with fiber-degrading microbes, such as those from Lachnospiracea, Ruminococcaceae, and Prevotella. Our study shows a clear difference in the microbiome between the stomach and colon of Japanese macaques in both composition and function. This study provides a preliminary look at the alpha diversity and taxonomic composition within the stomach microbiome of Japanese macaques, a hindgut-fermenting nonhuman primate.

In bisexual groups, dominant males occupy the central part of the group, which is advantageous because it is reproductively beneficial. However, high-ranking males do not necessarily monopolize reproductive success, which indicates that low-ranking males increase their reproductive success through alternative mating strategies. To reveal the effectiveness and cost of these strategies employed by group males, it is necessary to clarify the spatial configuration of males and their group, and show how males combine strategies in different situations. This study demonstrates the spatial configuration of male Japanese macaques (Macaca fuscata yakui) and their group when males adopt three mating strategies: mate guarding, sneak mating, and cross-boundary mating (mating with females of another group). High-ranking males tended to perform mate guarding, but they also mated with females in other groups when there were few sexually active females in their group. Low-ranking males performed sneak mating; they remained in the central part of the group to monitor sexually active females and moved toward the periphery for copulation but never completely left the group. In addition, males individually ventured outside the group's ranging area and succeeded in mating with females of other groups. The cross-boundary mating strategy has the advantage of increasing the number of potential mating partners, which is not present in the other two strategies. However, because of considerable costs and low contribution to mating frequency, this strategy is complementary and is employed when the expectation of mating success in their group is low.

Foregut fermentation is well known to occur in a wide range of mammalian species and in a single bird species. Yet, the foregut microbial community of free-ranging, foregut-fermenting monkeys, that is, colobines, has not been investigated so far. We analysed the foregut microbiomes in four free-ranging proboscis monkeys (Nasalis larvatus) from two different tropical habitats with varying plant diversity (mangrove and riverine forests), in an individual from a semi-free-ranging setting with supplemental feeding, and in an individual from captivity, using high-throughput sequencing based on 16S ribosomal RNA genes. We found a decrease in foregut microbial diversity from a diverse natural habitat (riverine forest) to a low diverse natural habitat (mangrove forest), to human-related environments. Of a total of 2700 bacterial operational taxonomic units (OTUs) detected in all environments, only 153 OTUs were shared across all individuals, suggesting that they were not influenced by diet or habitat. These OTUs were dominated by Firmicutes and Proteobacteria. The relative abundance of the habitat-specific microbial communities showed a wide range of differences among living environments, although such bacterial communities appeared to be dominated by Firmicutes and Bacteroidetes, suggesting that those phyla are key to understanding the adaptive strategy in proboscis monkeys living in different habitats.

Fecal DNA-based 16S ribosomal RNA (rRNA) gene sequencing using next-generation sequencers allows us to understand the dynamic gut microbiome adaptation of animals to their specific habitats. Conventional techniques of fecal microbiome analysis have been developed within the broad contexts defined by human biology; hence, many of these techniques are not immediately applicable to wild nonhuman primates. In order to establish a standard experimental protocol for the analysis of the gut microbiomes of wild animals, we selected the Japanese macaques (Macaca fuscata yakui) on Yakushima Island. We tested different protocols for each stage of fecal sample processing: storage, DNA extraction, and choice of the sequencing region in the bacterial 16S rRNA gene. We also analyzed the gut microbiome of captive Japanese macaques as the control. The comparison of samples obtained from identical macaques but subjected to different protocols showed that the tested storage methods (RNAlater and lysis buffer) produced effectively the same composition of bacterial operational taxonomic units (OTUs) as the standard frozen storage method, although the relative abundance of each OTU was quantitatively affected. Taxonomic assignment of the detected bacterial groups was also significantly affected by the region being sequenced, indicating that sequencing regions and the corresponding polymerase chain reaction (PCR) primer pairs for the 16S rRNA gene should be carefully selected. This study improves the current standard methods for microbiome analysis in wild nonhuman primates. Japanese macaques were shown to be a suitable model for understanding microbiome adaptation to various environments.

To expand our understanding of fission–fusion behavior and determine its variability among primates, studies of both individual-based and group-based fission–fusion are necessary. We conducted a parallel tracking study of male and female Japanese macaques (Macaca fuscata yakui) during the non-mating season to clarify the general features of separate ranging by males of this species, an example of fission–fusion behavior, and to reveal its associated costs and benefits. Males frequently engaged in short-term separate ranging, leaving the company of females and ranging on their own for periods averaging 68 min in duration. However, the males did not venture outside the group's home range. When ranging separately from the group, males spent more time feeding, particularly on fruit, stayed longer in each feeding tree, and fed at a lower rate than when ranging with the group. These behavioral changes suggest that males can avoid within-group feeding competition by ranging alone. However, this behavior was also associated with higher traveling costs, and these separated males were more vulnerable to intergroup competition and had fewer opportunities for social interaction. The frequency of separate ranging was lower when highly clumped food plant species were the main food source. Lower-ranked males, who received more aggression when ranging with the group, exhibited a higher frequency of separate ranging. This behavioral flexibility with respect to group cohesion may allow males to reduce the costs of group living without completely losing the benefits. Specifically, by ranging alone, males may acquire sufficient feeding time without being disturbed by other group members. Conversely, when ranging with the group, males can access grooming partners and advantages in intergroup competition. Am. J. Primatol. 76:374–384, 2014. © 2013 Wiley Periodicals, Inc.

Background: Gross motor development is usually assessed in terms of age of achievement of motor milestones. Although we generally have an impression that one developed faster, if one achieved younger milestone faster than usual; there are no longitudinal study about these association among each milestones especially in Japan. As a part of Japan Children's Study, we researched whether the achievement of gross motor milestones in infancy were related with the age of walking. Methods: This is a prospective cohort study of 290 healthy and term infants born in a district of Osaka City, Japan. Three milestones; rolling over, sitting and crawling were observed in laboratory at the age of 4 and 9 months by a pediatrician and a developmental psychologist, and the age of walking was confirmed in questionnaires filled in by the parents at the age of 18 and 27 months. Results: Whether or not roll over at 4 months, and sit and crawl at 9 months were related to the age of walking. About crawling and sitting, the age of walking varies not only able or unable but the variation of its form at the 9 month observation point. Multiple regression analysis revealed these three milestones were positively associated with walking; rolling over (β= 0.567), sitting (β= 1.973) and crawling (β= 1.473). Conclusion: The age and the patterns of sitting, crawling and rolling over were all related to the age of independent walking among Japanese infants. Consideration of milestone' definition and variations proves to be essential in medical check-up. Language: en

Digestion is an important process in understanding the feeding ecology of animals. We examined digesta passage time, digestibility, and total gut fill in Japanese macaques (Macaca fuscata; n = 4) under 4 dietary conditions representing the seasonal and regional variations in the diets of wild populations to determine the effects of food type and food intake on these digestive features. Food type is associated with mean retention time (MRT), digestibility, and total gut fill. Dry matter intake (DMI) of food correlates positively with total gut fill but not with MRT or digestibility. Conversely, indigestible DMI affected MRT negatively. Thus, when Japanese macaques consume high-fiber foods, MRT becomes shorter and digestibility is lower than when eating low-fiber foods. Moreover, macaques experience increases in total gut fill when they consume high-fiber diets or a large amount of food. Japanese macaques may excrete difficult-to-digest food components quickly; they nevertheless buffer an increase in food intake by an increase in gut fill. Our study offers new insights into the relationship between feeding ecology and nutritional physiology in primates by simultaneously examining the effects of food type and intake level on MRT and digestibility.

Background: Head circumference (HC) trajectories are held to reflect neurological development and the acquirement of intelligence. It is important to assess HC growth accurately because atypical HC growth is an indicator of various developmental disorders. HC growth is determined by both familial and physical factors but, hitherto, no one has considered both factors together. The aim of the present study was thus to investigate the relationship between HC, physical growth, and parental HC. Methods: The study group in the Japan Children's Study consisted of 192 healthy full-term Japanese children. HC (maximum occiptofrontal circumference), height and bodyweight were measured at the ages of 4, 9 and 18 months. Multiple regression analysis were conducted predicting the HC from the body measurements and mid-parental HC (defined as the average of standardized paternal and maternal HC). Results: Adjusted multiple R2 were 0.336, 0.307 and 0.259, measured at the aforementioned three stages. Bodyweight and mid-parental HC predicted the HC on each occasion (P < 0.01). Bodyweight was more relevant than mid-parental HC. Conclusions: HC growth is influenced by physical growth and parental HC; therefore, it is important to consider both physical and familial factors. A formula is herein proposed to assess HC using bodyweight and mid-parental HC.

We report the draft genome sequences of Sarcina ventriculi strains 14 and 17, both isolated from feces of wild Yakushima macaques ( Macaca fuscata yakui ). These genomic sequences will be helpful for the phylogenetic consideration of the family Clostridiaceae and understanding of the contribution of intestinal microbiota to the survival of Yakushima macaques.

Mycophagy (fungus-feeding) by Japanese macaques (Macaca fuscata yakui) in Yakushima has been observed by many researchers, but no detailed information is available on this behavior, including which fungal species are consumed. To provide a general description of mycophagy and to understand how and whether macaques avoid poisonous fungi, we conducted behavioral observation of wild Japanese macaques in Yakushima and used molecular techniques to identify fungal species. The results indicate that the diet of the macaques contains a large variety of fungal species (67 possible species in 31 genera), although they compose a very small portion of the total diet (2.2 % of annual feeding time). Fungi which were eaten by macaques immediately after they were picked up were less likely to be poisonous than those which were examined (sniffed, nibbled, carefully handled) by macaques. However, such examining behaviors did not appear to increase the macaques’ abilities to detect poisonous fungi. Fungi that were only partially consumed included more poisonous species than those fully consumed with/without examining behavior, yet this was not significant. Taste, therefore, might also play an important role in discriminating poisonous from non-poisonous.

Objective: Recently, disclosure of cancer diagnosis is common in Japan, but significant variability in patient preference of prognostic disclosure poses difficult questions for doctors. The aim of this study was to understand the reasons for wanting or not wanting to know life prognosis and how the information was interpreted and utilized by the patients. Methods: The study was based on narrative interviews with 42 women with breast cancer and 49 men with prostate cancer, in varying stages. A qualitative and interpretive approach was taken, combining thematic analysis with constant comparison. Results: While some of the participants voluntarily asked for prognosis to prepare themselves for the end of life, others were shocked by unexpected and unilateral disclosure. Some obtained prognostic information from books and websites. Some preferred to remain unaware of life prognosis, partly because they feared it would become a self-fulfilling prophecy. Discussion: The major problem underlying the practice of prognostic disclosure is the absence of mutual understanding of how such information will be utilized. These findings affirm that it should be used to empower patients to participate in the decision-making process.

Gross motor development is usually assessed in terms of age of achievement of motor milestones. Although there is generally an impression of faster development if the milestones are achieved at younger ages, no longitudinal studies have been done on the associations between the milestones, especially in Japan. As a part of the Japan Children's Study, the purpose of the present study was to determine whether the achievement of gross motor milestones in infancy is related with the age of walking.This was a prospective cohort study of 290 healthy and term infants born in a district of Osaka City, Japan. Three milestones (rolling over, sitting, and crawling) were observed in the laboratory for infants aged at 4 and 9 months by a pediatrician and a developmental psychologist, and the age of walking was confirmed in questionnaires filled in by the parents at 18 and 27 months.Children who could roll over at 4 months, and sit and crawl at 9 months, walked earlier than children who could not roll over, sit and crawl, respectively. With regard to crawling, children who were creeping had a 1 month delay in walking, and those who could not move forward had a 2 month delay compared to typical crawlers. On multiple regression analysis these three milestones were positively associated with walking: rolling over (β= 0.567), sitting (β= 1.973) and crawling (β= 1.473).The age and the patterns of sitting, crawling and rolling over were all related to the age of independent walking among Japanese infants. Consideration of milestone definition and variations is essential in medical check-up.

This study investigated the process involved in the lateralization of movements during infancy by observing upper extremity movements in a laboratory setting. Reaching for flying rings, balls, mini toy cars, and small round cookies were observed and recorded by videotape at 4 and 9 months of age. The subjects were 202 infants who participated in Japan Children's Study, a cohort study on the development of sociability. Infants reached for objects significantly more frequently at 9 months (98%) than at 4 months (40%) (p0.001). Though the lateral preference in reaching for balls at 4 months was ambiguous, reaching for toy cars was performed more frequently with the right hand at 9 months (50%) than with the left one (19%) (p0.01). Lateralization of the upper extremity movements is thought to appear by 9 months.

Head circumference (HC) trajectories are held to reflect neurological development and the acquirement of intelligence. It is important to assess HC growth accurately because atypical HC growth is an indicator of various developmental disorders. HC growth is determined by both familial and physical factors but, hitherto, no one has considered both factors together. The aim of the present study was thus to investigate the relationship between HC, physical growth, and parental HC.The study group in the Japan Children's Study consisted of 192 healthy full-term Japanese children. HC (maximum occiptofrontal circumference), height and bodyweight were measured at the ages of 4, 9 and 18 months. Multiple regression analysis were conducted predicting the HC from the body measurements and mid-parental HC (defined as the average of standardized paternal and maternal HC).Adjusted multiple R(2) were 0.336, 0.307 and 0.259, measured at the aforementioned three stages. Bodyweight and mid-parental HC predicted the HC on each occasion (P0.01). Bodyweight was more relevant than mid-parental HC.HC growth is influenced by physical growth and parental HC; therefore, it is important to consider both physical and familial factors. A formula is herein proposed to assess HC using bodyweight and mid-parental HC.