Researcher basic information

• Ph.D., Tohoku University, Mar. 2020
• Master, Tohoku University, Mar. 2017

• https://researchmap.jp/kokinishihara?lang=en

Research activity information

• Nov. 2023, JDS Communications, Editor's Choice
  Cultivation of enteroids from fresh and cryopreserved bovine duodenal tissues

• Dynamics of Blood Taurine Concentration and its Correlation with Nutritional and Physiological Status during the Fattening Period of Japanese Black Cattle
  Shuntaro Takai; Huseong Lee; Minji Kim; Shinichiro Torii; Nishihara Koki; Joonpyo Oh; Tatsunori Masaki; Kentaro Ikuta; Eiji Iwamoto; Kota Masuda; Yoshinobu Uemoto; Fuminori Terada; Satoshi Haga; Sanggun Roh
  Journal of Animal Science, Oxford University Press (OUP), 13 Nov. 2024
  Scientific journal, Abstract
  
  Taurine, biosynthesized from methionine or cysteine in the liver, plays a crucial regulatory role in bile acid conjugation, antioxidant effects, and glucose and cholesterol metabolism. This may influence the metabolic changes associated with fat accumulation in beef cattle. However, the physiological role of taurine in this species has not been fully elucidated. In this study, we explored the physiological role of taurine in Japanese Black steers (Bos taurus) in different phases during the fattening period. To examine the correlation among plasma taurine concentrations, various physiological parameters, and genes related to taurine synthesis in the liver, we used biopsied liver tissues, blood samples, and rumen fluids collected from 21 steers at three different stages, i.e., early (T1; 13 months of age), middle (T2; 20 months of age), and late (T3; 28 months of age) phases. Additionally, to investigate the regulatory mechanisms underlying the expression profile of taurine synthesis genes, primary bovine hepatocytes obtained from 4-week-old Holstein calves were treated with palmitate, oleate, acetate, propionate, or β-hydroxybutyrate (BHBA). Plasma taurine and cholesterol concentrations significantly (P < 0.001) increased in the T2 phase, which is potentially attributable to increased energy intake and assimilation induced by increased intake of concentrated feed. Cysteine sulfinic acid decarboxylase (CSAD) expression significantly increased (P < 0.01) in T2 than in other phases. The expression levels of cysteine dioxygenase type 1 (CDO1) and cholesterol 7 alpha-hydroxylase (CYP7A1) were significantly higher (P < 0.001) in T2 than in T3; moreover, the CDO1/glutamate-cysteine ligase catalytic subunit (GCLC) ratio was higher (P < 0.05) in T2 than in T1. Plasma taurine concentrations were positively correlated with plasma methionine (r = 0.51; P < 0.05) and total cholesterol (r = 0.56; P < 0.05) concentrations at T2. Relative CDO1 mRNA expression was upregulated in cultured bovine hepatocytes treated with oleate and propionate, whereas it was downregulated upon acetate treatment. These findings indicate that the increase in plasma taurine concentrations in the T2 phase is associated with changes in lipid and methionine metabolism in Japanese Black steers.
• Preweaning Megasphaera elsdenii Supplementation in Dairy-Beef Calves: Impact on Performance, Behavior, and Rumen Development
  G. Mazon; J.M.V. Pereira; K. Nishihara; M.A. Steele; J.H.C. Costa
  Journal of Dairy Science, American Dairy Science Association, Oct. 2024, [Peer-reviewed]
  Scientific journal
• Bacteria colonization and gene expression related to immune function in colon mucosa is associated with growth in neonatal calves regardless of live yeast supplementation.
  Koki Nishihara; Clothilde Villot; Lautaro Cangiano; Le Luo Guan; Michael Steele
  Journal of Animal Science and Biotechnology, 15, 1, 76, 76, 05 Jun. 2024, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, BACKGROUND: As Holstein calves are susceptible to gastrointestinal disorders during the first week of life, understanding how intestinal immune function develops in neonatal calves is important to promote better intestinal health. Feeding probiotics in early life may contribute to host intestinal health by facilitating beneficial bacteria colonization and developing intestinal immune function. The objective of this study was to characterize the impact of early life yeast supplementation and growth on colon mucosa-attached bacteria and host immune function. RESULTS: Twenty Holstein bull calves received no supplementation (CON) or Saccharomyces cerevisiae boulardii (SCB) from birth to 5 d of life. Colon tissue biopsies were taken within 2 h of life (D0) before the first colostrum feeding and 3 h after the morning feeding at d 5 of age (D5) to analyze mucosa-attached bacteria and colon transcriptome. Metagenome sequencing showed that there was no difference in α and β diversity of mucosa-attached bacteria between day and treatment, but bacteria related to diarrhea were more abundant in the colon mucosa on D0 compared to D5. In addition, qPCR indicated that the absolute abundance of Escherichia coli (E. coli) decreased in the colon mucosa on D5 compared to D0; however, that of Bifidobacterium, Lactobacillus, and Faecalibacterium prausnitzii, which could competitively exclude E. coli, increased in the colon mucosa on D5 compared to D0. RNA-sequencing showed that there were no differentially expressed genes between CON and SCB, but suggested that pathways related to viral infection such as "Interferon Signaling" were activated in the colon mucosa of D5 compared to D0. CONCLUSIONS: Growth affected mucosa-attached bacteria and host immune function in the colon mucosa during the first 5 d of life in dairy calves independently of SCB supplementation. During early life, opportunistic pathogens may decrease due to intestinal environmental changes by beneficial bacteria and/or host immune function. Predicted activation of immune function-related pathways may be the result of host immune function development or suggest other antigens in the intestine during early life. Further studies focusing on the other antigens and host immune function in the colon mucosa are required to better understand intestinal immune function development.
• Assessing the impact of three feeding stages on rumen bacterial community and physiological characteristics of Japanese Black cattle.
  Huseong Lee; Minji Kim; Tatsunori Masaki; Kentaro Ikuta; Eiji Iwamoto; Koki Nishihara; Itoko Nonaka; Akane Ashihara; Youlchang Baek; Sungdae Lee; Yoshinobu Uemoto; Satoshi Haga; Fuminori Terada; Sanggun Roh
  Scientific reports, 14, 1, 4923, 4923, 28 Feb. 2024, [International Magazine]
  English, Scientific journal, In Japan, Japanese Black cattle, known for their exceptional meat quality owing to their abundant intramuscular fat, undergo a unique three-stage feeding system with varying concentrate ratios. There is limited research on physiological and rumen microbial changes in Japanese Black cattle during these stages. Therefore, this study aimed to examine Japanese Black steers in these three stages: early (T1, 12-14 months), middle (T2, 15-22 months), and late (T3, 23-30 months). The rumen bacteria of 21 cattle per phase was analyzed using 16S rRNA gene sequencing. Rumen bacterial diversity was significantly higher in T1, with a distinct distribution, than in T2 and T3. Specific phyla and genera were exclusive to each stage, reflecting the shifts in feed composition. Certain genera dominated each stage: T1 had Flexilinea, Streptococcus, Butyrivibrio, Selenomonas, and Kandleria; T2 had Bifidobacterium, Shuttleworthia, and Sharpea; and T3 had Acetitomaculum, Mycoplasma, Atopobium, and Howardella. Correlation analysis revealed significant associations between certain microbial populations and physiological parameters. These findings indicate that changes in energy content and feed composition are associated with physiological and ruminal alterations. This study may guide strategies to improve rumen health and productivity in Japanese Black cattle by modifying diets to specific fattening stages.
• TLR5 ligand induces the gene expression of antimicrobial peptides and CXCL8 through IL-1β gene expression in cultured rumen epithelial cells.
  Koki Nishihara; Yutaka Suzuki; Satoshi Haga; Sanggun Roh
  Animal science journal = Nihon chikusan Gakkaiho, 95, 1, e13972, 2024, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, High grain feeding or weaning, which could compromise the rumen epithelium by increasing ruminal short-chain fatty acid (SCFA) concentrations with pH reduction, is associated with high levels of ruminal toll-like receptor 5 (TLR5). This study aimed to determine the role of TLR5 in the rumen epithelium. Immunohistochemistry revealed that TLR5 was localized in cells on the basal side (i.e., basal and spinous layers) rather than in the granular layer in the rumen epithelium, where tight junctions are most potent, in pre- and post-weaning calves (n = 9). Primary bovine rumen epithelial cells (BRECs) obtained from Holstein cows (n = 3) were cultured to investigate the factors that upregulate TLR5; however, SCFA, low pH (pH 5.6), BHBA, L-lactate, D-lactate, and LPS did not upregulate TLR5 gene expression in BREC. Primary BREC treated with flagellin (TLR5 ligand) had higher expression of interleukin-1β (IL-1β) (P < 0.05) than BREC treated with vehicle. In addition, BREC treated with IL-1β had higher expression of antimicrobial peptides and C-X-C motif chemokine ligand 8 than BREC treated with vehicle (P < 0.05). These results suggest that ruminal TLR5 may recognize epithelial disruption via flagellin and mediate the immune response via IL-1β during high-grain feeding or weaning.
• Cultivation of enteroids from fresh and cryopreserved bovine duodenal tissues
  Koki Nishihara; Katie M. Wood; Le Luo Guan; Michael A. Steele
  JDS Communications, 4, 6, 518, 523, American Dairy Science Association, Aug. 2023, [Peer-reviewed], [Lead author]
  Scientific journal
• Reduction in mucosa thickness is associated with changes in immune function in the colon mucosa during the weaning transition in Holstein bull dairy calves.
  Koki Nishihara; Jolet van Niekerk; Zhixiong He; David Innes; Le Luo Guan; Michael Steele
  Genomics, 115, 5, 110680, 110680, 15 Jul. 2023, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, This study aims to characterize changes in the structure and the molecules related to immune function in the colon mucosa in dairy calves during the weaning transition (weaned at week 6 of age). Colon mucosa thickness, measured at week 5 to 8 and 12 of age, decreased for 2 weeks after weaning, but then recovered. Colon mucosa's transcriptome profiling at week 5, 7, and 12 of age was obtained using RNA-sequencing. Functional analysis showed that pathways related to immune function were up-regulated postweaning. A weighted gene co-expression network analysis identified 17 immune function related genes, expressed higher postweaning, which were negatively correlated with colon mucosa thickness, suggesting that these genes may be involved in colon mucosa inflammation and recovery from mucosa thickness decrement during the weaning transition. As such, it is important to determine the function of immune cells in the colon mucosa during the weaning transition in dairy calves.
• Physiological roles and regulation of hepatic angiopoietin-like protein 3 in Japanese Black cattle (Bos taurus) during the fattening period.
  Rika Shikida; Minji Kim; Makoto Futohashi; Koki Nishihara; Huseong Lee; Yutaka Suzuki; Yeolchang Baek; Tatsunori Masaki; Kentaro Ikuta; Eiji Iwamoto; Yoshinobu Uemoto; Satoshi Haga; Fuminori Terada; Sanggun Roh
  Journal of animal science, 15 Jun. 2023, [International Magazine]
  English, Scientific journal, Angiopoietin-like protein 3 (ANGPTL3) is expressed predominantly in the liver and plays a major role in regulating the circulating triglyceride and lipoprotein fraction concentrations by inhibiting lipoprotein lipase (LPL) activity. Given these physiological roles, ANGPTL3 may play an important role in metabolic changes related to fat accumulation during the fattening period in Japanese Black. This study aimed to reveal the physiological roles of hepatic ANGPTL3 in Japanese Black steers (Bos taurus) during the fattening period and investigate the regulatory effects of hepatic ANGPTL3. To investigate the gene expression and protein localization of ANGPTL3, 18 tissue samples were collected from tree male Holstein bull calves aged 7 weeks. Biopsied liver tissues and blood samples were collected from 21 Japanese Black steers during the early (T1; 13 months of age), middle (T2; 20 months), and late fattening phases (T3; 28 months). Relative mRNA expression, blood metabolite concentrations, hormone concentrations, growth, and carcass traits were analyzed. To identify the regulatory factors of hepatic ANGPTL3, primary bovine hepatocytes collected by 2 Holstein calves aged 7 weeks were incubated with insulin, palmitate, oleate, propionate, acetate, or beta-hydroxybutyric acid (BHBA). The ANGPTL3 gene was most highly expressed in the liver, with minor expression in the renal cortex, lungs, reticulum, and jejunum in Holstein bull calves. In Japanese Black steers, relative ANGPTL3 mRNA expressions were less as fattening progressed, and blood triglyceride, total cholesterol, and non-esterified fatty acid (NEFA) concentrations increased. Relative ANGPTL8 and Liver X receptor alpha (LXRα) mRNA expressions decreased in late and middle fattening phases, respectively. Furthermore, relative ANGTPL3 mRNA expression was positively correlated with ANGPTL8 (r = 0.650; P < 0.01) and ANGPTL4 (r = 0.540; P < 0.05) in T3 and T1, respectively, and LXRα showed no correlation with ANGPTL3. Relative ANGTPL3 mRNA expression was negatively correlated with total cholesterol (r = -0.434; P < 0.05) and triglyceride (r = -0.645; P < 0.01) concentrations in T3 and T1, respectively; There was no significant correlation between ANGTPL3 and carcass traits. Relative ANGTPL3 mRNA expression in cultured bovine hepatocytes was downregulated in oleate treatment. Together, these findings suggest that ANGPTL3 downregulation in late fattening phases is associated with the changes in lipid metabolism.
• Transcriptome profiling revealed that key rumen epithelium functions change in relation to short-chain fatty acids and rumen epithelium-attached microbiota during the weaning transition.
  Koki Nishihara; Jolet van Niekerk; David Innes; Zhixiong He; Angela Cánovas; Le Luo Guan; Michael Steele
  Genomics, 115, 5, 110664, 110664, 05 Jun. 2023, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, This study aims to characterize the functional changes of the rumen epithelium associated with ruminal short-chain fatty acid (SCFA) concentration and epithelium-attached microbes during the weaning transition in dairy calves. Ruminal SCFA concentrations were determined, and transcriptome and microbiota profiling in biopsied rumen papillae were obtained from Holstein calves before and after weaning using RNA- and amplicon sequencing. Metabolic pathway analysis showed that pathways related to SCFA metabolism and cell apoptosis were up- and down-regulated postweaning, respectively. Functional analysis showed that genes related to SCFA absorption, metabolism, and protective roles against oxidative stress were positively correlated with ruminal SCFA concentrations. The relative abundance of epithelium-attached Rikenellaceae RC9 gut group and Campylobacter was positively correlated with genes involved in SCFA absorption and metabolism, suggesting that these microbes can cooperatively affect host functions. Future research should examine the contribution of attenuated apoptosis on rumen epithelial functional shifts during the weaning transition.
• Characteristics of Physiological Parameters of Japanese Black Calves Relate to Carcass Weight
  Shotaro Arakawa; Minji Kim; Tatsuya Aonuma; Michihiro Takagi; Satoshi Watanabe; Huseong Lee; Koki Nishihara; Satoshi Haga; Yoshinobu Uemoto; Sanggun Roh
  Animals, 13, 3, 487, 487, MDPI AG, 31 Jan. 2023
  Scientific journal, This study aimed to identify the growth performance and blood factors associated with carcass weight in Japanese Black calves based on 675 performance tests and field carcass records. We measured the body weight, withers height, and chest girth at the start of fattening age (approximately 8–10 months) and analyzed eight blood factors, including vitamins and metabolites. Single- and two-trait animal models were used to estimate the heritability and genetic correlations. The heritability estimates for growth performance were moderate to high (ranging from 0.48 to 0.74), and those for blood metabolites were low to moderate (ranging from 0.19 to 0.51). Estimates for genetic correlations of carcass or body weight with body weight, withers height, and chest girth were high (ranging from 0.42 to 0.80). The body weight and withers height at 8 months of age are possibly closely related to the final carcass weight. The blood metabolites associated with body weight were vitamin E in steers (castrated males) and β-carotene in heifers. Our findings indicate that body measurements and blood metabolites measured during the growing period could be used to determine the nutritional and physiological status of cattle as well as predict carcass weight.
• Physiological responses and adaptations to high methane production in Japanese Black cattle
  Minji Kim; Tatsunori Masaki; Kentaro Ikuta; Eiji Iwamoto; Koki Nishihara; Makoto Hirai; Yoshinobu Uemoto; Fuminori Terada; Sanggun Roh
  Scientific Reports, 12, 1, Springer Science and Business Media LLC, Jul. 2022
  Scientific journal, Abstract
  
  In this study, using enteric methane emissions, we investigated the metabolic characteristics of Japanese Black cattle. Their methane emissions were measured at early (age 13 months), middle (20 months), and late fattening phases (28 months). Cattle with the highest and lowest methane emissions were selected based on the residual methane emission values, and their liver transcriptome, blood metabolites, hormones, and rumen fermentation characteristics were analyzed. Blood β-hydroxybutyric acid and insulin levels were high, whereas blood amino acid levels were low in cattle with high methane emissions. Further, propionate and butyrate levels differed depending on the enteric methane emissions. Hepatic genes, such as SERPINI2, SLC7A5, ATP6, and RRAD, which were related to amino acid transport and glucose metabolism, were upregulated or downregulated during the late fattening phase. The above mentioned metabolites and liver transcriptomes could be used to evaluate enteric methanogenesis in Japanese Black cattle.
• Chemerin Regulates Epithelial Barrier Function of Mammary Glands in Dairy Cows
  Yutaka Suzuki; Sachi Chiba; Koki Nishihara; Keiichi Nakajima; Akihiko Hagino; Won-Seob Kim; Hong-Gu Lee; Tomonori Nochi; Toru Suzuki; Sang-Gun Roh
  Animals, 11, 11, 3194, 3194, MDPI AG, 09 Nov. 2021
  Scientific journal, Epithelial barrier function in the mammary gland acts as a forefront of the defense mechanism against mastitis, which is widespread and a major disorder in dairy production. Chemerin is a chemoattractant protein with potent antimicrobial ability, but its role in the mammary gland remains unelucidated. The aim of this study was to determine the function of chemerin in mammary epithelial tissue of dairy cows in lactation or dry-off periods. Mammary epithelial cells produced chemerin protein, and secreted chemerin was detected in milk samples. Chemerin treatment promoted the proliferation of cultured bovine mammary epithelial cells and protected the integrity of the epithelial cell layer from hydrogen peroxide (H2O2)-induced damage. Meanwhile, chemerin levels were higher in mammary tissue with mastitis. Tumor necrosis factor alpha (TNF-α) strongly upregulated the expression of the chemerin-coding gene (RARRES2) in mammary epithelial cells. Therefore, chemerin was suggested to support mammary epithelial cell growth and epithelial barrier function and to be regulated by inflammatory stimuli. Our results may indicate chemerin as a novel therapeutic target for diseases in the bovine mammary gland.
• Ruminal epithelial insulin‐like growth factor‐binding proteins 2, 3, and 6 are associated with epithelial cell proliferation
  Koki Nishihara; Yutaka Suzuki; Sanggun Roh
  Animal Science Journal, 91, 1, Wiley, Jan. 2020, [Peer-reviewed], [Lead author]
  Scientific journal
• ルーメン上皮の恒常性維持機構
  西原 昂来; 盧 尚建
  栄養生理研究会報, 63, 2, 27, 34, Sep. 2019, [Peer-reviewed], [Invited], [Lead author]
• Growth of rumen papillae in weaned calves is associated with lower expression of insulin-like growth factor-binding proteins 2, 3, and 6.
  Koki Nishihara; Yutaka Suzuki; Dahye Kim; Sanggun Roh
  Animal Science Journal, 90, 9, 1287, 1292, Sep. 2019, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, This study aimed to characterize the relationship between the growth of rumen papillae in calves and the mRNA expression of insulin-like growth factor-binding proteins (IGFBPs) in the rumen papillae. The length of rumen papillae, the mRNA expression of IGFBPs in rumen papillae by quantitative real-time PCR, and the presence of insulin-like growth factors I and II (IGF-I and II) by immunohistochemistry (IHC) were analyzed in nine Holstein calves divided into three groups: suckling (2 weeks, n = 3), milk-continued (8 weeks, n = 3), and weaned (8 weeks, n = 3). The length of rumen papillae was greater (p < 0.01) in weaned calves than in suckling and milk-continued calves, whereas the expressions of IGFBP2, IGFBP3, and IGFBP6 genes were lower (p < 0.05) in the rumen papillae of weaned calves than in milk-continued calves. Thus, rumen papillae length and IGFBP2, 3, and 6 expressions were negatively correlated. The IHC analysis showed that IGF-I and IGF-II were present in the rumen epithelium of calves. These results suggested that the growth of rumen papillae after weaning is associated with the induction of IGFs by the low levels of IGFBP2, IGFBP3, and IGFBP6.
• Effects of butyrate supplementation in antibiotic-free milk replacer and starter on growth performance in suckling calves.
  Sanggun Roh; Nobuhiro Kimura; Kazushi Sakamoto; Koki Nishihara; Keiichi Suzuki; Kazuo Katoh
  Animal Science Journal, 89, 10, 1486, 1491, Oct. 2018, [Peer-reviewed], [International Magazine]
  English, Scientific journal, The aim of this study was to evaluate butyrate supplementation of antibiotic-free milk replacer and starter on growth performance in male Holstein calves. Twenty-nine calves were divided into two groups. Group C (n = 13) was fed antibiotic-free milk replacer without supplementation, and Group B (n = 16) was fed antibiotic-free milk replacer supplemented with butyrate (1.6 % DM of Gustor BP70® ). Starter in Group B contained 0.3 % DM of Gustor BP70® . The intake of milk replacer was lower in group B than in C (p = 0.07 for the treatment x week interaction). Body weight (BW) and heart girth (HG) in group B was higher than in C during the experimental period (p = 0.07 and 0.01 for the treatment × week interaction, respectively). The duration of the weaning period in group B was shorter than in group C (p = 0.02). β-hydroxybutyrate (BHBA) was higher in group B than in C (p = 0.04). Insulin like growth factor-1 (IGF-1) concentrations tended to be higher in group B than in C (p = 0.07 for treatment × week interaction). Our results show that butyrate supplementation in antibiotic-free milk replacer and starter exerted positive effects on growth performance in suckling calves.
• Downregulated angiopoietin-like protein 8 production at calving related to changes in lipid metabolism in dairy cows.
  Misato Nakano; Yutaka Suzuki; Satoshi Haga; Eri Yamauchi; Dahye Kim; Koki Nishihara; Keiichi Nakajima; Takafumi Gotoh; Seungju Park; Myunggi Baik; Kazuo Katoh; Sanggun Roh
  Journal of Animal Science, 96, 7, 2646, 2658, 29 Jun. 2018, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Acute physiological adaptation of lipid metabolism during the postpartum transition period of cows facilitates peripheral metabolic regulation. Hepatokines, which are hormones secreted from hepatocytes, are presumed to play a critical role in systemic metabolic regulation. Angiopoietin-like protein 8 (ANGPTL8) has been identified as a novel hepatokine associated with circulating triglyceride concentrations in mice and humans. However, regulation of ANGPTL8 and its physiological effects is still unknown in cattle. The present study aimed to reveal changes in ANGPTL8 expression and secretion during the periparturient period, and to investigate its regulatory effect on adipocytes and mammary epithelial cells. In the peripartum period, liver ANGPTL8 mRNA expression was lesser on the day of parturition and 1 wk postpartum than it was 1 wk before parturition (P < 0.05). Moreover, plasma ANGPTL8 concentrations decreased on the day of parturition as compared with that 1 wk before parturition (P < 0.05). In addition, ANGPTL8 expression in cultured bovine hepatocytes was downregulated after oleate and palmitate treatment but upregulated after insulin treatment (P < 0.05). ANGPTL8 decreased hormone-sensitive lipase (HSL) expression in differentiated adipocytes and cluster of differentiation 36 (CD36), fatty acid synthase (FAS), acetyl-coa carboxylase (ACC), and stearoyl-coa desaturase (SCD) in cultured bovine mammary epithelial cells (P < 0.05). These data suggest that hepatic ANGPTL8 production was downregulated postpartum when the cows experienced a negative energy balance. This downregulation was associated with increased concentrations of NEFA and decreased concentrations of insulin in lactating cows, and it facilitated lipid mobilization from adipose tissue to the mammary glands. We speculate that ANGPTL8 might have beneficial effects in reverting or improving the physiological adaptation and pathological processes of lipid metabolism during the peripartum period.
• Comparative transcriptome analysis of rumen papillae in suckling and weaned Japanese Black calves using RNA sequencing.
  Koki Nishihara; Daichi Kato; Yutaka Suzuki; Dahye Kim; Misato Nakano; Yu Yajima; Satoshi Haga; Miwa Nakano; Hiroshi Ishizaki; Ryouka Kawahara-Miki; Tomohiro Kono; Kazuo Katoh; Sang-Gun Roh
  Journal of Animal Science, 96, 6, 2226, 2237, 04 Jun. 2018, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, The length and density of rumen papillae starts to increase during weaning and growth of ruminants. This significant development increases the intraruminal surface area and the efficiency of VFA (acetate, propionate, butyrate, etc.) uptake. Thus, it is important to investigate the factors controlling the growth and development of rumen papillae during weaning. This study aimed to compare the transcriptomes of rumen papillae in suckling and weaned calves. Total RNA was extracted from the rumen papillae of 10 male Japanese Black calves (5 suckling calves, 5 wk old; 5 weaned calves, 15 wk old) and used in RNA-sequencing. Transcript abundance was estimated and differentially expressed genes were identified and these data were then used in Ingenuity Pathway Analysis (IPA) to predict the major canonical pathways and upstream regulators. Among the 871 differentially expressed genes screened by IPA, 466 genes were upregulated and 405 were downregulated in the weaned group. Canonical pathway analysis showed that "atherosclerosis" was the most significant pathway, and "tretinoin," a derivative of vitamin A, was predicted as the most active upstream regulator during weaning. Analyses also predicted IgG, lipopolysaccharides, and tumor-necrosis factor-α as regulators of the microbe-epithelium interaction that activates rumen-related immune responses. The functional category and the up-regulators found in this study provide a valuable resource for studying new candidate genes related to the proliferation and development of rumen papillae from suckling to weaning Japanese Black calves.
• Post-prandial decrease in plasma growth hormone levels is not related to the increase in plasma insulin levels in goats.
  Koki Nishihara; Ryoko Kobayashi; Yutaka Suzuki; Katsuyoshi Sato; Kazuo Katoh; Sanggun Roh
  Asian-Australasian journal of animal sciences, 30, 12, 1696, 1701, Dec. 2017, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, OBJECTIVE: In the present study, we examined whether the post-prandial reduction in plasma growth hormone (GH) levels is related to the increase in plasma insulin levels in ruminants. METHODS: We performed two experiments: intravenous bolus injection of insulin (0.2 IU/kg body weight) or glucose (1.0 mmol/kg body weight) was administered to increase the plasma insulin levels in male Shiba goats. RESULTS: In the insulin injection experiment, significant (p<0.05) increase in GH concentrations was observed, 15 to 20 min after the injection; it was accompanied with a significant (p<0.01) increase in cortisol concentrations at 45 to 90 min, when compared to the concentrations in the saline-injected controls. The glucose injection significantly (p<0.05) increased the plasma GH concentration at 20 to 45 min; this was not accompanied by significantly higher cortisol concentrations than were observed for the saline-injected control. Hypoglycemia induced by the insulin injection, which causes the excitation of the adrenal cortex, might be involved in the increase in insulin levels. CONCLUSION: Based on these results, we conclude that post-prandial increases in plasma insulin or glucose levels do not induce a decrease in GH concentration after feeding in the ruminants.

• Effects of housing environment on intake, growth, and physiological responses of dairy calves during summer
  R. Fukami; K. Nishihara; K. Murayama; N. Kobayashi; T. Sugino; M. Oba
  American Dairy Science Association, Jun. 2026, English, Poster presentation
• 〔Major achievements〕Associations among average daily gain, activity, and ambient temperature–humidity index in Holstein heifer calves during summer and fall.
  K. Nishihara, R. Fukami, K. Murayama, N. Kobayashi, T. Sugino, and M. Oba
  American Dairy Science Association, Jun. 2026, English, Poster presentation
• 〔主要な業績〕Bacteria colonization and gene expression related to immune function in colon mucosa is associated with growth in neonatal calves.
  K. Nishihara; C. Villo; L. R. Cangiano; L. L. Guan; M. Steele
  American Dairy Science Association Annual Meeting, Jun. 2024, English
• The effect of prepartum dietary starch on colostrum fatty acid content in Holstein dairy cattle.
  L. E. Engelking; A. J. Fischer-Tlustos; J. Haisan; W. Shi; J. Yu; K.Nishihara; M. Oba; M. A. Steel
  American Dairy Science Association Annual Meeting, Jun. 2024, English
• Saccharomyces cerevisiae boulardii CNCM I-1079 influence on gut permeability, intestinal microbiota, and host immune function in newborn dairy calves.
  S. Jantzi; K. Nishihara; C. Villo; L. Rostoll Cangiano; M. Steele
  American Dairy Science Association Annual Meeting, Jun. 2023, English
• Effects of a Megasphaera elsdenii oral capsule on reticulorumen volatile fatty acid dynamics and papillae development in dairy-beef calves.
  G. Mazon; J. M. V. Pereira; K. Nishihara; M. A. Steele; J. H. C. Costa
  American Dairy Science Association Annual Meeting, Jun. 2023, English
• 〔Major achievements〕RNA-sequencing analysis of biopsied rumen papillae revealed that key rumen epithelium functions change in relation to short-chain fatty acids and rumen epithelium-attached microbiota during the weaning transition in dairy calves
  Koki Nishihara; Jolet van Niekerk; David Innes; Zhixiong He; Angela Cánovas; Le Luo Guan; Michael Steele
  American Dairy Science Association Annual Meeting, Jun. 2023
• 〔Major achievements〕Reduction in the colon mucosa thickness during the weaning transition is associated with molecular changes involved in immune function in dairy calves
  Koki Nishihara, Jolet van Niekerk, Le Luo Guan, Michael Steele
  American Dairy Science Association Annual Meeting, Jun. 2023
• 〔Major achievements〕Cryopreservation and resuscitation of bovine duodenum tissues and enteroids cultivation methods
  Koki Nishihara; Katie M. Wood; Le Luo Guan; Michael A. Steele
  American Dairy Science Association Annual Meeting, Jun. 2023, English
• The influence of transition diet energy and protein content on colostrum and early lactation milk composition and bioactive compound concentrations in Holstein dairy cattle
  Amanda J. Fischer-Tlustos; M. Victoria Sanz-Fernandez; Dave J. Seymour; Koki Nishihara; Justin B; Renaud, Joh; P. Cant; Michael A. Steele
  The Animal Nutrition Conference of Canada, May 2022
• マルコフ連鎖モデルを用いた肉用牛繁殖農家における繁殖マネジメントの評価
  西原 昂来; 藪内 祐樹
  日本畜産学会第129回大会, Sep. 2021
• Roles of Toll-Like Receptor 5 Ligand in bovine rumen epithelial cells
  Nishihara K; Roh SG
  3rd International Symposium on Immunology and Biotechnology, Nov. 2019, English, Invited oral presentation
• Roles of Toll-like receptor 5 ligand in the innate immune system in primary bovine rumen epithelial cells
  Nishihara K; Futohashi M; Roh S
  International Symposium on Ruminant Physiology, Sep. 2019, English, Oral presentation
• CCL5–monocyte/macrophage axis maintains homeostasis in rumen epithelium
  Nishihara K; Haga S; Roh SG
  American Society of Animal Science – Canadian Society of Animal Science Annual Meeting, Jul. 2018, English, Poster presentation
• Ussing チャンバー法を用いた亜急性ルーメンアシドーシス時のルーメン上皮崩壊モデルの構築
  西原 昂来; 太箸 誠; 盧 尚建
  日本畜産学会第124回大会, Mar. 2018
• ウシ第１胃の絨毛における生体防御とケモカインの関係
  西原 昂来; 鈴木 裕; 芳賀 聡; 盧 尚建
  生命科学系学会合同年次大会, Dec. 2017
• Rumen Epithelial Cells as a source of Inflammatory Cytokines
  Nishihara K; Suzuki Y; Haga S; Kim DH; Roh, SG
  Joint Symposium of Japan-China-Korea on Rumen Metabolism and Physiology, Oct. 2017, English
• ウシルーメン絨毛組織の発達におけるIGFとIGFBPsの役割
  西原 昂来; 鈴木裕; 金 多慧; 中野 美智; 中野 美和; 石崎 宏; 芳賀 聡; 盧 尚建
  日本畜産学会第122回大会, Mar. 2017, Oral presentation
• The roles of IGF-1 and IGFBPs on the developments of rumen papillae
  Nishihara K; Suzuki Y; Roh SG
  International Animal Sciences and Biotechnology Forum, Dec. 2016, English
• ウシのルーメン発達関連因子の解析
  西原 昂来; 加藤 大地; 鈴木 裕; 芳賀 聡; 川原 玲香; 河野 友宏; 盧 尚建
  生物資源ゲノム解析拠点シンポジウム, Sep. 2016
• 反芻動物の採食に伴う成長ホルモン分泌抑制に関する研究
  小林 涼子; 西原 昂来; 盧 尚建; 加藤 和雄
  日本畜産学会第119回大会, Mar. 2015

• 北海道における母胎内暑熱ストレスが後継牛に与える影響の解明
  若手研究人材育成事業 若手研究人材・ネットワーク育成補助金（タレント補助金）
  Jul. 2026 - Mar. 2027
  西原昂来
  ノーステック財団, Principal investigator
• 食肉製品の色調改善に寄与する細菌の遺伝子発現解析
  Apr. 2026 - Mar. 2027
  西原昂来; 若松純一
  公益財団法人 伊藤記念財団, Principal investigator
• The study about controling rumen epithelial barrier function to prevent the disruption of rumen epithelium
  Grants-in-Aid for Scientific Research Grant-in-Aid for JSPS Fellows
  25 Apr. 2019 - 31 Mar. 2021
  西原 昂来
  濃厚飼料といった易発酵性飼料の多給は、亜急性ルーメンアシドーシス(SARA)といった生産病を招くことがある。これまでの研究より、培養ルーメン上皮細胞においてTLR5（Toll様受容体 5）リガンドを添加するとIL-1β（インターロイキン-1β）の発現量が高くなり、そのIL-1βはDEFB1といった抗菌ペプチドの発現を亢進することが示唆された。よって、今年度は、以下１、２を実施した。
  １、ウシルーメン上皮細胞における抗菌ペプチドの発現調節の解明：前年度の実験より、TLR5リガンドはルーメン上皮細胞においてDEFB1といった抗菌ペプチドの発現を亢進することが示唆された。その他の抗菌ペプチドとしてS100タンパク質ファミリーのTLR5リガンドの影響を検討したところ、培養ルーメン上皮細胞においてS100A8とS100A9の発現が亢進した。また、LPS（リポポリサッカロイド）の添加によってもS100A8とS100A9の発現が亢進した。また、LPSはIL-1βの発現も亢進した。
  ２、ウシルーメン上皮細胞におけるIL-1βの役割の解明：これまでの研究により、TLR5リガンドの添加により培養ルーメン上皮細胞においてIL-1βの発現が高くなることが分かっている。培養ルーメン上皮細胞の増殖においてIL-1βの影響を検討したところ、IL-1βの添加により、培養ルーメン上皮細胞の増殖速度が有意に高くなった。
  以上の結果から、ウシのルーメン上皮細胞のバリア機能が崩壊すると、TLR5リガンドやLPSなどによって、抗菌ペプチドやIL-1βの発現が亢進し、微生物侵入の阻止や細胞増殖促進されることが示唆された。これらの機構が、上皮の修復・保護を担うと考えられる。
  Japan Society for the Promotion of Science, Grant-in-Aid for JSPS Fellows, Tohoku University, 19J12823
• 子ウシの健全な発達を目指した消化管の上皮細胞接着微生物叢の相互関係の解明
  研究助成
  Apr. 2020 - Mar. 2021
  西原昂来
  伊藤記念財団, Principal investigator