Supplemental Clostridium butyricum MIYAIRI 588 Affects Intestinal Bacterial Composition of Finishing Pigs.

Animal gastrointestinal tracts are populated by highly diverse and complex microbiotas. The gut microbiota influences the bioavailability of dietary components and is closely associated with physiological processes in the host. Clostridium butyricum reportedly improves growth performance and affects the gut microbiota and immune functions in post-weaning piglets. However, the effects of C. butyricum on finishing pigs remain unclear. Therefore, we herein investigated the effects of C. butyricum MIYAIRI 588 (CBM588) on the gut microbiota of finishing pigs. 16S rRNA gene sequencing was performed using fecal samples and ileal, cecal, and colonic contents collected after slaughtering. The α-diversity of the small intestinal microbiota was lower than that of the large intestinal microbiota, whereas β-diversity showed different patterns depending on sample collection sites. The administration of CBM588 did not significantly affect the α- or β-diversity of the microbiotas of fecal and intestinal content samples regardless of the collection site. However, a linear discriminant ana-lysis Effect Size revealed that the relative abundance of Lactobacillaceae at the family level, Bifidobacterium at the order level, and Lactobacillus ruminis and Bifidobacterium pseudolongum at the species level were higher in the fecal samples and cecal and colonic contents of the treatment group than in those of the control group. Therefore, the administration of CBM588 to finishing pigs affected the composition of the gut microbiota and increased the abundance of bacteria that are beneficial to the host. These results provide important insights into the effects of probiotic administration on relatively stable gut microbial ecosystems.


Diet-Induced Thermogenesis and Expression Levels of Thyroid Hormone Target Genes and Their Products in Rats Differ between Meat Proteins.

We compared the effects of purified meat proteins on postprandial thermogenesis and on the secretion of and responsiveness to thyroid hormones (THs) in rats. Body temperatures at 2 h after feeding were significantly higher in the chicken and mutton protein groups than in the other groups, and these proteins seem to have a strong thermogenic effect. There were no significant differences in plasma TH concentrations among the groups, but levels of TH-responsive Spot 14 protein in the liver and brown adipose tissue were significantly higher in the chicken and mutton protein groups than in the other groups. Levels of malic enzyme 1 protein in the liver and brown adipose tissue were significantly higher in the chicken protein group than in the other groups except for the mutton protein group. Furthermore, levels of uncoupling protein 1 were higher in the chicken and mutton protein groups than in the other groups. The results suggest that the difference in postprandial thermogenesis of meat is strongly dependent on meat proteins; chicken and mutton proteins are strong promoters of postprandial thermogenesis, and THs may contribute to this effect. Since strong postprandial thermogenesis and high expression levels of TH target genes and their products were not observed in the amino acid group, chicken and mutton proteins or their digested peptides might contribute to these effects.

Journal of Nutritional Science and Vitaminology


The validation of the NH Immunochromato O157 test kit.

NH Immunochromato O157 is an immunochromatographic test for detection of Escherichia coli O157:H7 and O157:NM in food. It enables simple and rapid testing for the target organism after 18-24 h enrichment. In inclusivity and exclusivity testing, all 50 O157:H7 strains and 15 O157:NM strains tested positive, while all 33 exclusivity strains yielded negative results. Taken together, all 98 strains tested in inclusivity/exclusivity testing were identified correctly. NH Immunochromato O157 method was compared to U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook, Chapter 5.08, reference method for detection of E. coli O157:H7 in 25 g of raw ground beef. The performance of both methods was revealed to be statistically equivalent. Autoclaved and non-autoclaved sample enrichments yielded the same results, showing sterilization is not mandatory for testing with NH Immunochromato O157. The results of the robustness study were not statistically different in all conditions, suggesting that NH Immunochromato produce reliable results under various conditions. However, the users are recommended to follow the instruction when applying sample to the test strip, because a smaller sample volume may produce invalid result.


Effect of ear-corn silage feeding on meat productivity of fattening pigs

イアコーンサイレージ給与が肥育豚の産肉性に及ぼす影響

The production and use of ear-corn silage (ECS) have been studied to further promote utilization of self-supplied animal feed in Hokkaido. The purpose of this study was to evaluate the effects of ECS feeding on the meat productivity of fattening pigs. Fattening pigs (an average body weight of 65.4kg) were divided into 3 groups with 6 animals in each group : the control group fed with "commercial compound feed alone," the ECS group fed with mixed feed consisting of "80% commercial compound feed+20% ECS (dry matter)" and the ECEFS group fed with mixed feed consisting of "60% compound feed+20% ECS+20% eco-feed silage (EFS) consisting primarily of starch pulp (dry matter)." All pigs were fattened until their average body weight reached 117.9kg. Feed conversion ratio (Feed intake/Gain) was similar in the control and ECS groups, and was significantly higher in the ECEFS group (P < 0.05) at total period. No significant differences were found in all of the meat quality parameters except skatole. Fat level of skatole was significantly lower in the ECS and ECEFS group (P < 0.05). These results indicate that replacement of 20% ECS as dry matter has comparable productivity to no replacement control, and feeding of ECS to fattening pigs may reduce unpleasant skatole smell of produced meat. Our study suggests that ECS can be useful as alternative feedstuffs for fattening pigs.

自給飼料活用の更なる推進のため，北海道ではイアコーンサイレージ(ECS)の利用検討が行なわれている．本研究では，ECS給与が肥育豚の産肉性に及ぼす影響を検討した．肥育豚(平均65.4kg)を6頭ずつ3区分し，市販配合飼料を給与した対照区，配合飼料80%+ECS 20%の混合飼料を給与したECS区，配合飼料60%+ECS 20%+デンプン粕主体エコフィードサイレージ(EFS)20%の混合飼料を給与したECEFS区を設け，平均117.9kgまで肥育した．飼料(乾物)要求率は，対照区とECS区では同等で，ECEFS区では全期間平均で有意に高かった(P < 0.05)．各試験区の肉質は，背部皮下脂肪中のスカトール含量が，対照区と比較してECS区，ECEFS区で有意に低かった(P < 0.05)．以上から，20%量のECSを配合飼料と置換しても生産性に影響なく，産肉の不快臭を減少させる可能性が示唆された．


Promising Nucleic Acid Lateral Flow Assay Plus PCR for Shiga Toxin-Producing Escherichia coli.

Shiga toxin (Stx)-producing Escherichia coli (STEC) is a frequent cause of foodborne infections, and methods for rapid and reliable detection of STEC are needed. A nucleic acid lateral flow assay (NALFA) plus PCR was evaluated for detecting STEC after enrichment. When cell suspensions of 45 STEC strains, 14 non-STEC strains, and 13 non-E. coli strains were tested with the NALFA plus PCR, all of the STEC strains yielded positive results, and all of the non-STEC and non-E. coli strains yielded negative results. The lower detection limit for the STEC strains ranged from 0.1 to 1 pg of genomic DNA (about 20 to 200 CFU) per test, and the NALFA plus PCR was able to detect Stx1- and Stx2-producing E. coli strains with similar sensitivities. The ability of the NALFA plus PCR to detect STEC in enrichment cultures of radish sprouts, tomato, raw ground beef, and beef liver inoculated with 10-fold serially diluted STEC cultures was comparable to that of a real-time PCR assay (at a level of 100 to 100,000 CFU/ml in enrichment culture). The bacterial inoculation test in raw ground beef revealed that the lower detection limit of the NALFA plus PCR was also comparable to that obtained with a real-time PCR assay that followed the U.S. Department of Agriculture guidelines. Although further evaluation is required, these results suggest that the NALFA plus PCR is a specific and sensitive method for detecting STEC in a food manufacturing plant.


Mechanisms and consequences of carnosine-induced activation of intestinal epithelial cells


Identification of anti-allergic lactic acid bacteria that supress Ca2+ influx and histamine release in human basophilic cells


Survey of content of trans-fatty acids in meat

Japanese Journal of Food Chemistry and Safety


Detection of soybean proteins in fermented soybean products by using heating extraction.

Soybean is used in processed foods worldwide. Because soybean can cause adverse reactions in some atopic patients, appropriate labeling regarding its content in processed foods is needed to better protect consumers. In the previous study, we developed a reliable sandwich Enzyme Linked Immunosorbent Assay (ELISA) method with high sensitivity and specificity for detecting soybean proteins by using antibody to Gly m Bd 30K, which was originally characterized as a vacuolar protein with a molecular mass of 34 kDa in soybean. The ELISA displayed satisfactory repeatability and reproducibility in an interlaboratory evaluation. However, it could not detect soybean protein in fermented soybean products. We therefore developed an extraction method combined with a heating process to inhibit soybean protein degradation by microbial proteolytic enzymes in fermented soybean products. This extraction method enables the sensitive detection of soybean protein in fermented soybean products such as natto and miso. It was able to detect with high-sensitivity soybean protein present at 10 μg/g levels in model processed foods. This method is suitable for quantifying soybean protein in processed foods without the degrading effects of microbial proteolytic enzymes. The present extraction method can be used sensitively to monitor labeling systems in a reliable manner and should be useful for the mandatory inspections required under Japanese regulations. The extraction and ELISA methods that we developed enable sensitive detection of soybean protein in soybean products, including fermented foods. These methods should be useful for reliable and sensitive monitoring of product labeling systems and should help to solve the problem of insensitive in soybean labeling of processed foods.


Effects of oral administration of heat-killed Enterococcus faecium strain NHRD IHARA in post-weaning piglets.

Probiotic bacteria such as lactic acid bacteria (LAB) have recently received attention as candidates for alternative anti-microbial feed additives. We previously isolated Enterococcus faecium strain NHRD IHARA (FERM BP-11090, NHRD IHARA strain) and reported its probiotic efficacy. However, we have not determined the effect of oral administration of heat-killed cells of this strain. Here, we performed two experiments to investigate the effect of oral administration of the heat-killed NHRD IHARA strain on post-weaning piglets. In Experiment 1, there was a significant improvement in growth performance (P = 0.04) and increase in serum immunoglobulin A (IgA) production (P = 0.03) in the group fed heat-killed cells. These results were similar to previous results we obtained with live cells. We also found changes in serum and fecal IgA production that were unrelated to the patterns of microbiotal change. In Experiment 2, we detected a significant improvement in villus growth in the jejunum (P = 0.0002). In conclusion, oral administration of the heat-killed NHRD IHARA strain in post-weaning piglets had the same efficacy as administration of the live strain. The heat-killed NHRD IHARA strain can be used as feed additives to improve pig growth and health on commercial farms.


Multiple marker effects of single nucleotide polymorphisms in three genes, AKIRIN2, EDG1 and RPL27A, for marbling development in Japanese Black cattle.

Marbling in beef, measured by Beef Marbling Standard (BMS) number, is an economically important trait for beef cattle breeding and markets in Japan. We previously detected three single nucleotide polymorphisms (SNPs) associated with BMS number of Japanese Black in Oita prefecture: c.*188G>A in AKIRIN2, g.1471620G>T in EDG1 and g.3109537C>T in RPL27A. Here, we carried out single and multiple marker association analyses for the three SNPs in a different commercial Japanese Black population of 892 genotyped animals. The single marker analyses with the model including a single SNP showed significant associations of all SNPs with BMS number. The multiple marker analysis with the model including the main effects of the three SNPs and their interactions detected only significant main effects of g.1471620G>T and g.3109537C>T and a significant interaction between c.*188G>A and g.1471620G>T. These findings suggest the presence of inter-allelic interactions among genes affecting the development of beef marbling. For effective marker-assisted selection for BMS number, interactions among these markers need to be considered.


Improved Functional Properties of Potato Starch through Simultaneous Heat-moisture Treatment and Conjugation with Lysine and Fatty Acylated Saccharide by the Maillard Reaction


Postprandial thermic effect of chicken involves thyroid hormones and hepatic energy metabolism in rats.

We investigated the postprandial thermic effect of chicken and its mechanisms in rats. A chicken diet showed a strong thermic effect after consumption, and the removal of fat induced more rapid and stronger thermogenesis. Although thermogenesis induced by a purified chicken protein diet was also strong, the thermic reaction was not so rapid and a remarkable rise of peripheral temperatures was not observed. Defatted chicken and purified chicken protein activated the thyroid hormone system and up-regulated rate-limiting enzyme genes of glucose metabolism and the tricarboxylic acid (TCA) cycle in the liver. Moreover, chicken protein up-regulated the mRNA expression of a rate-limiting enzyme of hepatic lipid metabolism. It is possible that the mechanisms by which body temperature is raised are different between chicken protein and defatted chicken. On the other hand, it is possible that chicken fat suppressed the expression of energy metabolism-related genes that was induced by the consumption of lean chicken. As a result, a rise of postprandial body temperature might not have been induced after consumption of chicken fat. These results suggest that the consumption of lean chicken activates the thyroid hormone system and hepatic energy metabolism and consequently induces the postprandial thermic effect of chicken.

Journal of Nutritional Science and Vitaminology


Development of an immunochromatographic test strip for detection of cholera toxin.

Because cholera toxin (CT) is responsible for most of the symptoms induced by Vibrio cholerae infection, detection of CT is critical for diagnosis of the disease. In this study, we constructed an immunochromatographic test strip for detection of CT (CT-IC) with polyclonal antibodies developed against purified recombinant whole CT protein. The detection limit of the CT-IC was 10 ng/mL of purified recombinant CT, and it could detect the CT in culture supernatant of all 15 toxigenic V. cholerae isolates examined, whereas no false-positive signal was detected in all 5 nontoxigenic V. cholerae isolates examined. The specificity of the CT-IC was examined with recombinant heat-labile toxin (LT), which shares high homology with CT, and it was revealed that the minimum detection limit for LT was 100 times higher than that for CT. In addition, lt gene-positive enterotoxigenic Escherichia coli (ETEC) was examined by CT-IC. The false-positive signals were observed in 3 out of 12 ETEC isolates, but these signals were considerably faint. The CT-IC did not develop false-positive signals with all 7 V. parahaemolyticus isolates. These results showed the high specificity of CT-IC and the feasible use of it for the detection and surveillance of toxigenic V. cholerae.


Characterization of probiotic properties of Enterococcus faecium NHRD IHARA isolated from porcine feces.

We examined in vitro the adhesion of Enterococcus faecium NHRD IHARA (NHRD IHARA) to porcine small intestinal mucin (PSIM) and inhibition of the adherence of enteropathogenic bacteria due to pre-incubation of PSIM with NHRD IHARA. NHRD IHARA exhibited an effective barrier function in porcine small intestinal mucus layer.

Bioscience, Biotechnology, and Biochemistry


Comparative Analyses of Anti-hypertensive Effects of Commercial Collagen Peptides in Spontaneously Hypertensive Rats

市販コラーゲンペプチドの自然発症性高血圧ラットにおける血圧上昇抑制作用

In this study, we investigated the anti-hypertensive effects of two enzymatic hydrolysates of chicken and pig collagens in spontaneously hypertensive rats (SHR). SHR were divided into three groups and administrated collagen peptides at 3 g/kg body weight, or water for 14 weeks. In the chicken collagen peptides group, systolic blood pressure was significantly reduced when compared with the control group, and diastolic blood pressure was also reduced when compared to the pig collagen group. Microscopic analysis revealed that inflammation in liver and heart was mildly ameliorated in the chicken collagen peptides group.

ニワトリ由来コラーゲンペプチド CCOP およびブタ由来コラーゲンペプチド P-LAP を自然発症性高血圧ラット SHR に 14 週間連続摂取させる長期飼育試験を実施した．その結果，SHR の収縮期血圧において，CCOP 摂取群は，対照群に比べて 8 週目および 14 週目で，P-LAP 摂取群に比べて 14 週目に有意な低値を示した．また拡張期血圧においても，CCOP 摂取群は P-LAP 摂取群に比べて 14 週目に有意な低値を示した．一方，対照群と P-LAP 摂取群の間の血圧値にはいずれも有意な変化は認められなかった．また，病理組織観察の結果，CCOP 摂取群は対照群に比べて，肝臓および心臓において緩やかな臓器保護現象が認められた．


Development of a novel multiplex lateral flow assay using an antimicrobial peptide for the detection of Shiga toxin-producing Escherichia coli.

The binding capacity of peptides with broad antimicrobial activity, or antimicrobial peptides (AMPs), to microbes has recently been applied to the specific detection of bacteria and viruses. We established a novel lateral flow assay (LFA) that combines AMPs labeled with colloidal gold and a target-specific antibody immobilized on a nitrocellulose membrane. α-Helical AMPs, especially cecropin P1 (CP1), magainin 2 (MG2), and ceratotoxin A (CtxA), were shown to have optimal properties as probes in LFA. We also established a multiplex LFA for the simultaneous detection and identification of three serogroups of Shiga toxin-producing Escherichia coli (STEC) using the CP1 probe with polyclonal antibodies anti-O157, anti-O26, and anti-O111. Each serogroup of E. coli could easily and rapidly be detected by multiplex LFA using CP1 and each was clearly visualized in a different position on the LFA strip. The multiplex LFA could detect all tested E. coli strains from serogroups O157 (22/22), O26 (17/17), and O111 (7/7), and the detection limit was 10(4)CFU/mL. No other serogroups of E. coli, including STEC O45, O91, O103, O121, and O145, or non-E. coli strains, reacted. The multiplex LFA could detect E. coli O157, O26, and O111 in food samples at very low levels (6.3, 2.9, and 5.6 CFU per 25 g of ground beef, respectively) after 18-h enrichment, and these results were in accordance with the results of the culture method, immunochromatography (IC) strip, and PCR. Given the broad binding capacity, AMP probes in combination with specific antibodies in the novel multiplex LFA may have the potential to detect various microbes simultaneously with identification on a single strip.


Potential rapid and simple lateral flow assay for Escherichia coli O111.

We developed and evaluated a lateral flow assay (LFA) as a simple and rapid method for direct detection of Escherichia coli O111 in food after enrichment. When cell suspensions of 8 E. coli O111 strains and 77 non-E. coli O111 strains were tested with the LFA, the former all yielded positive results and the latter all yielded negative results. The minimum detection limits for the E. coli O111 strains were 1.8 × 10(3) to 5.6 × 10(5) CFU/ml of cell suspension, and the LFA was able to detect live cultures or those killed by autoclaving at nearly the same level of sensitivity. To evaluate the ability of LFA to detect its target in food, enrichment cultures of meat samples inoculated with 10-fold serial dilutions of E. coli O111 were tested with the LFA and PCR. Even when there were very few E. coli O111 cells in the meat samples (1.6 × 10(0) to 1.6 × 10(1) CFU/25 g of food), when they were cultured in modified E. coli broth with novobiocin for 22 h at 42°C, the LFA yielded positive results that corresponded to the PCR results. Although the LFA requires further evaluation and field study, these results suggest that this assay has sufficient sensitivity and specificity. This procedure can be completed with a one-step incubation after the test strip has been inserted into the sample after 22 h of culture, whereas the standard culture method requires multiple cultures, skilled personnel, a well-equipped laboratory, and 4 or 5 days. The speed and simplicity of this LFA make it suitable for use as part of routine screening assays in the food industry.


豚の解体処理工程におけるPseudomonas属菌の汚染状況

豚の解体処理工程におけるPseudomonas属菌の汚染状況

Journal of the Japan Veterinary Medical Association


Effects of a chicken collagen hydrolysate on the circulation system in subjects with mild hypertension or high-normal blood pressure.

We investigated the effects of a chicken collagen hydrolysate (CCH) on the circulation system in humans. A total of 58 subjects with either mild hypertension (systolic blood pressure (SBP) of 140-159 mmHg or diastolic blood pressure (DBP) 90-99 mmHg) or high-normal blood pressure (SBP 130-139 mmHg or DBP 85-89 mmHg) were assigned to two groups, one involving a placebo and the other, the test food (including CCH of 2.9 g/d). The parameters related to each subject's circulation system were monitored over the study period of 18 weeks. The Δbrachial-ankle pulse wave velocity (baPWV), an indicator of arterial stiffness and marker of vascular damage, was significantly lower in the test food group than in the placebo group during the treatment period. The blood pressure in the test food group was also significantly lower than that in the placebo group, while the serum nitrogen oxide was higher in the test food group after the treatment. These results suggest that CCH exerted modulatory effects on the human circulation system.


Simple, rapid, and reliable detection of Escherichia coli O26 using immunochromatography.

Shiga toxin-producing Escherichia coli (STEC) O26 has been increasingly associated with diarrheal disease all over the world. We developed an immunochromatographic (ic) strip for the rapid detection of E. coli O26 in food samples. To determine the specificity of the IC strip, pure cultures of 67 E. coli and 22 non-E. coli strains were tested with the IC strip. The IC strip could detect all (18 of 18) E. coli O26 strains tested and did not react with strains of any other E. coli serogroup or non-E. coli strains tested (0 of 71). The minimum detection limits for E. coli O26 were 2.2 × 10(3) to 1.0 × 10(5) cfu/ml. To evaluate the ability of the IC strip to detect E. coli O26 in food, 25-g food samples (ground beef, beef liver, ground chicken, alfalfa sprout, radish sprout, spinach, natural cheese, and apple juice) were spiked with E. coli O26. The IC strip was able to detect E. coli O26 at very low levels (approximately 1 cfu/25 g of food samples) after an 18-h enrichment, and the IC strip results were in 100% agreement with the results of the culture method and pcr assay. When 115 meat samples purchased from supermarkets were tested, 5 were positive for E. coli O26 with the IC strip; these results were confirmed with a pcr assay. These results suggest that the IC strip is a useful tool for detecting E. coli O26 in food samples.


Dietary animal proteins alter monoamine metabolism in the brain.

Several amino acids have effects on mental function, including sedative, antidepressant-like and anxiolytic-like effects. However, the influence of integrated amino acid nutrition as protein constituents on mental function remains unclear. Therefore, the purpose of the present study was to compare the influence of chicken, pork and beef protein extracts on brain monoamine metabolism in mice. Changes in monoamine levels and their turnover rates in the brain were induced by different protein sources. In particular, chicken protein group showed the highest norepinephrine levels in the hippocampus and hypothalamus, and beef protein extract caused an activation of the serotonergic system in the hypothalamus, although there were no significant differences in amino acid compositions of these protein extracts. Therefore, it was revealed that amino acid compositions in dietary protein did not induce alteration in monoamine metabolism. However, there were differences in small molecular peptides, such as creatine, carnosine and anserine levels in animal protein extracts. In conclusion, monoamine metabolism was altered by dietary protein sources. However, it was indicated that the alteration in monoamine metabolism may be independent from amino acid compositions in dietary protein. In addition, alteration in monoamine metabolism depending on the dietary protein sources may be induced by small molecular peptides.


Expression profiles of carnosine synthesis-related genes in mice after ingestion of carnosine or ß-alanine.

Carnosine is a dipeptide that improves exercise performance. The carnosine synthesis mechanism through carnosine and ß-alanine ingestion remains unclear. Therefore, we investigated the tissue distribution of carnosine synthase, ATP-grasp domain-containing protein-1 (ATPGD1) mRNA, and ATPGD1 and carnosine specific dipeptidase (CN1) gene expression profiles in mice that were given carnosine or ß-alanine orally. ddY mice (7-week-old) were randomly divided into three groups (n = 6 to 8 animals per group) and were orally given 2 g/kg body weight of carnosine, ß-alanine, or water. After 15, 30, 60, 120, 180, or 360 min of treatment, the tissues (brain, blood, liver, kidneys, olfactory bulbs, hindleg muscles) were collected. The obtained tissues measured the expression of ATPGD1 and CN1 genes using quantitative PCR methods. The ATPGD1 gene was expressed in muscle and to a lesser extent in brain. The expression of ATPGD1 in the vastus lateralis muscle increased significantly at 180 min (P = 0.023) after carnosine ingestion and 60 (P = 0.023) and 180 min (P = 0.025) after ß-alanine ingestion. Moreover, the carnosine group showed a significantly increased renal expression of the CN1 gene 60 min after ingestion (P = 0.0015). The ATPGD1 gene showed high expression levels in brain and muscle. The ß-alanine or carnosine administration significantly increased ATPGD1 and CN1 expression in mice.

Journal of the International Society of Sports Nutrition


Chicken collagen hydrolysate-derived peptides inhibit tumor necrosis factor-α-induced inflammatory response in endothelial cells


Impacts of acute imipramine treatment on plasma and brain amino acid metabolism in mice given graded levels of dietary chicken protein.

Several studies have shown a relationship between depression and animal protein intake. To evaluate whether the difference of dietary chicken protein levels induces an antidepressant-like effect and potentiates acute antidepressant effects, three levels of dietary chicken protein were used as the representative animal protein with imipramine used as the antidepressant. In addition, the effects of dietary chicken protein on brain metabolism were evaluated. Open field test (OFT) and forced swimming test (FST) were conducted on the 27th and 28th days, respectively. OFT and FST were not influenced by both imipramine and dietary protein levels. However, characteristic effects of imipramine treatment on brain monoamine metabolism were observed in the cerebral cortex and hypothalamus. In addition, dietary protein significantly increased taurine and L-ornithine levels even though these amino acids were not contained in the diets. In conclusion, the metabolism of several amino acids in the plasma and brain were altered by dietary chicken protein.


Effect of Long-term Ingestion of Chicken Extract on Muscular Power for Middle and Advanced Age Groups

鶏肉抽出物の摂取が中高齢者の筋力に及ぼす影響

The effects of carnosine and anserine rich chicken extract on muscular power and fatigue sensation in middle and advanced age groups were investigated. Twenty middle and advanced aged healthy volunteers were divided into two groups and administered chicken extract at 1 500 mg/day (225 mg/day as carnosine + anserine) or 0 mg/day for 4 weeks. Before and after the administration period, isokinetic knee extension power, flexion power, and one-leg standing time with eyes open were assessed. Chicken extract administration significantly increased knee extension power and one-leg standing time. These results suggest that carnosine- and anserine-rich chicken extract administration effectively improves muscular power in middle and advanced age.

カルノシン，アンセリンを高濃度に含有する鶏肉抽出物の摂取が，中高齢者の筋力にもたらす影響を検討した．中高齢者20名を2群に分け，鶏肉抽出物をそれぞれ一日量に1 500 mg(カルノシン，アンセリンの合計量として225 mg)もしくは0 mgを4週間継続摂取させた．摂取期間の前後に，等速性膝最大伸展力，膝最大屈曲力，および開眼片足立ちの保持時間を測定した．その結果，鶏肉抽出物群において，有意な膝最大伸展力の向上，開眼片足立ち保持時間の延長が認められた．これらの結果より，カルノシン，アンセリンを含有する鶏肉抽出物の摂取は，中高齢者の筋力の向上に有効であると考えられた．


Identification of Immunopotentiating Lactic Acid Bacteria that Induce Antibody Production by in vitro Stimulated Human Peripheral Blood Mononuclear Cells.

L-leucyl-L-leucine methyl ester (LLME) is known to remove lysosome-rich cells from human peripheral blood mononuclear cells (PBMCs). To evaluate the immunopotentiating ability of lactic acid bacteria (LAB), we adopted the in vitro stimulation protocol of LLME-treated PBMCs as a model assay system and monitored the level of antibody produced by stimulated PBMCs. The results indicated that several LAB strains have immunopotentiating ability against PBMCs, as evidenced by the enhanced antibody production and increased number of antigen-specific B cells. Next, we identified T cells as the direct target cells of the immunopotentiating LAB strain L32, suggesting that L32 induced antibody production by PBMCs through T-cell activation. Finally, we tested the immunopotentiating ability of ligands for Toll-like receptor 2 (TLR2), which is known to mediate the LAB signal, and observed that both L32 and one of the TLR2 ligands, LTA-BS, induced antigen-specific antibody production by in vitro stimulated PBMC. This suggests that L32 and LTA-BS can be used as an adjuvant for stimulating immune reaction in PBMCs.

Bioscience of Microbiota, Food and Health


Effect of liquid whey feeding on fecal microbiota of mature and growing pigs.

The effect of liquid whey feeding on fecal bacteria and their metabolites was assessed in five pregnant sows and 66 growing pigs. Sows were fed a control diet for 4 weeks (control period) followed by the same diet but with whey feeding (5 L/day/pig) for 4 weeks (whey period). One group of growing pigs was given 267 L of whey per pig (whey group), while the other group was not (control group). In both cases, liquid whey was given separately from control diet. Sows in the whey period had feces showing lower pH, lower ammonia concentration, and larger population sizes of total bacteria, lactobacilli and bifidobacteria. The bacterial gene library analysis indicated that Mitsuokella and Megasphaera were more frequently detected, while Clostridium disporicum were detected less frequently in the whey period. Feces from whey-fed growing pigs showed lower pH than that from control pigs in the early stage of growing. Also, larger populations of total bacteria, lactobacilli and bifidobacteria were recorded in the whey group. From the bacterial gene library analysis, the detection frequency of Lactobacillus reuteri tended to be higher in the whey group. These results indicate that whey feeding influences the hindgut microbiota of pigs, possibly leading to a fermentation shift that is favorable for animal health.


Possible Association of Single Nucleotide Polymorphism in Titin Gene with Growth-Related Trait in Japanese Black Beef Cattle

Journal of Animal and Veterinary Advances


軟骨抽出物の骨膜細胞への効果

軟骨抽出物の骨膜細胞への効果


Effect of Long-term Ingestion of Porcine Elastin Peptide on Human Skin Elasticity

ヒトにおけるブタ由来エラスチンペプチド摂取による皮膚弾力性向上作用

The effect of oral administration of elastin peptide (hydrolysate of elastin from porcine aorta) on human skin elasticity was studied. First, five healthy volunteers were administered elastin peptide after fasting for 12h. A significant increase in total amino acid concentration in blood was observed. The composition of increased amino acids in blood was similar to the amino acid composition of the elastin peptide. In addition, the concentrations of peptide forms of hydroxyproline and arginine significantly increased in blood. These results suggest that a portion of the administered elastin peptide migrates into blood in peptide form. Second, thirty-nine middle-aged volunteers were divided into three groups and administered 0, 100, or 200mg/day of elastin peptide for 8 weeks. Significant increases in skin elasticity were observed in the 100and 200-mg groups after 8 weeks. Increase in skin elasticity of the 200-mg group was significantly higher than that in the 0-mg group. These results suggest that ingestion of elastin peptide increases skin elasticity.

ブタ大動脈を原料としてエラスチン加水分解ペプチド(エラスチンペプチド)を調製し，その摂取がヒトの皮膚弾力性にもたらす影響について検討した．はじめに，エラスチンペプチド経口摂取後のヒト血液中のアミノ酸濃度の変化を観察した．成人男性5名を被験者として，12時間絶食後にエラスチンペプチドを摂取させた．その結果，エラスチンペプチド経口摂取後に血中の総アミノ酸量が増加し，増加したアミノ酸の組成は，摂取したエラスチンペプチドのアミノ酸組成に類似していた．また，ハイドロキシプロリンおよびアルギニンがそれぞれペプチド態として血中に検出され，エラスチンペプチドの少なくとも一部はペプチド態として血中に移行していると考えられた．次に，39名の中高齢者を3群にわけ1日量0, 100, 200mgのエラスチンペプチドを8週間継続摂取させ皮膚弾力性を測定した．100mg, 200mg摂取群において摂取開始8週目に摂取前と比較して有意に皮膚弾力性が上昇した．またその変化率は0mgと比較して200mg群で有意に高値を示した．以上より，エラスチンペプチドの経口摂取はヒトの皮膚弾力性を向上させることが示唆された．


Effect of liquid straight whey feeding on reproductive performance, productivity and carcass characteristics of pigs in a commercial pig farm

豚生産農場における液状ストレートホエイ給与が繁殖性, 生産性および枝肉形質に及ぼす影響

An enormous amount of whey is discharged from daily factories, and farms in which pigs are fed with whey are increasing because of its high protein content and nutritive value. However, effects of whey feeding have not been fully investigated. In the present study, we investigated the reproductive performance, the productivity and the carcass characteristics of pigs fed with commercial formula feed supplemented with liquid straight whey. Sows were fed with three to five L of liquid straight whey per day per head. Litter size, rate of return to estrus within seven days and conception rate of whey-fed sows were significantly higher than those of control sows : they improved by 0.2 heads (P < 0.001), 3.2% (P < 0.05) and 3.4% (P < 0.001), respectively, in whey-fed sows. One point three to 1.5 L per day per head of liquid straight whey was fed to fattening pigs. Shipping age of whey-fed pigs was significantly younger than that of control pigs (3.4 days, P < 0.05). Significant differences in the carcass characteristics between whey-fed and control pigs were not observed. Meanwhile, when 0.2 L per day per head of liquid straight whey was fed to pigs only in the weaning period, shipping age and daily gain were not improved in whey-fed pigs. Significant differences in the carcass characteristics were not observed either. These results showed that liquid straight whey feeding is effective in improving the reproductive performance of sows in a commercial pig farm. Improvement of productivity was also observed when whey was fed to fattening pigs.

ホエイは牛乳からナチュラルチーズを製造する際に大量に生成される．本試験では国内商用豚生産農場にて繁殖雌豚，哺乳期肉豚および肥育期肉豚に対して液状ストレートホエイを通常飼料に付加する形で一定量給与し，その繁殖成績や発育成績および枝肉形質に対する効果を検証した．繁殖雌豚へ1日3-5 Lの液状ストレートホエイ給与によって一腹産子数，離乳後7日以内再発情率および受胎率がそれぞれ有意に0.2頭，3.2%および3.4%改善した．肥育期肉豚に1日1.3-1.5 Lの液状ストレートホエイを給与した場合，出荷日齢が有意に3.4日短縮することが確認された．一方，肉豚の哺乳期のみ1日0.2 Lの液状ストレートホエイ給与ではその後の発育成績の改善は認められなかった．以上の結果から，商用豚生産農場において繁殖雌豚や肥育期の肉豚に適切な量の液状ストレートホエイを給与すれば繁殖成績や発育成績の改善が期待できることがわかった．


Establishment of a novel method of screening anti-allergic lactic acid bacteria.

In this study, we attempted to establish a novel method of screening anti-allergic lactic acid bacteria (LAB). We cloned the human histidine decarboxylase (HDC) promoter into the promoter-less pPhi-Yellow-RPL-dest1 vector and established KU812F cells transduced with this vector (pHDCp-Phi-Yellow/KU812F). After adding LAB to these cells, the change in fluorescence intensity was monitored by flow cytometry. After screening, we identified several LAB strains that downregulated HDC promoter activity. Functional analysis of these LAB strains indicated that two LAB strains inhibited histamine release from KU812F cells, indicating that this assay system can be used to screen for anti-allergic LAB in a high-throughput manner.

Bioscience, Biotechnology, and Biochemistry


Absorption and effectiveness of orally administered low molecular weight collagen hydrolysate in rats.

Collagen, a major extracellular matrix macromolecule, is widely used for biomedical purposes. We investigated the absorption mechanism of low molecular weight collagen hydrolysate (LMW-CH) and its effects on osteoporosis in rats. When administered to Wistar rats with either [(14)C]proline (Pro group) or glycyl-[(14)C]prolyl-hydroxyproline (CTp group), LMW-CH rapidly increased plasma radioactivity. LMW-CH was absorbed into the blood of Wistar rats in the peptide form. Glycyl-prolyl-hydroxyproline tripeptide remained in the plasma and accumulated in the kidney. In both groups, radioactivity was retained at a high level in the skin until 14 days after administration. Additionally, the administration of LMW-CH to ovariectomized stroke-prone spontaneously hypertensive rats increased the organic substance content and decreased the water content of the left femur. Our findings show that LMW-CH exerts a beneficial effect on osteoporosis by increasing the organic substance content of bone.

Journal of Agricultural and Food Chemistry


Simple and rapid detection of Campylobacter spp. in naturally contaminated chicken-meat samples by combination of a two-step enrichment method with an immunochromatographic assay.

A simple and rapid method to detect Campylobacter spp. in chicken-meat samples was established. This method consisted of a combination of a two-step enrichment method with a commercially available immunochromatographic assay, named NH Immunochromato Campylobacter (NH IC Campy, Nippon Meat Packers, Ibaraki, Japan), which is able to detect Campylobacter antigen in an enrichment culture within 15 min. The enrichment method did not require much blood or a particular system of generating a microaerobic atmosphere, in contrast to the standard method of enriching Campylobacter spp. in chicken-meat samples. The sensitivity of a combination of the two-step enrichment method with NH IC Campy for detection of non- and freeze-stressed Campylobacter spp. in spiked chicken meat was determined using bacterial cells of Campylobacter jejuni and Campylobacter coli. The detection sensitivities for non-stressed C. jejuni and C. coli were found to range from 5.5 to 1.3x10(1) CFU per 25 g of chicken meat, and those for freeze-stressed C. jejuni and C. coli were found to range from 9.2x10(1) to 1.5x10(2) CFU per 25 g of chicken meat. When a total of 68 chicken-meat samples were tested, the combination method determined that 61 samples were positive for Campylobacter spp. This method was more sensitive than a bacterial culture test, which consists of standard enrichment culturing and plating onto selective agars. Because the combination could be conducted in approximately 48 h, from the beginning of the enrichment culture to final determination, it was more rapid than the bacterial culture test, which requires four to five days. Moreover, the combination was simple to perform. These results suggest that combining the two-step enrichment method with NH IC Campy is useful as a simple and rapid alternative to the conventional bacterial culture test for detecting Campylobacter spp. in naturally contaminated chicken meat samples.

International Journal of Food Microbiology


Effects of exposure to hyperbaric oxygen on oxidative stress in rats with type II collagen-induced arthritis.

Arthritis was induced in 9-week-old female Dark Agouti rats by injecting type II collagen. Serum levels of the derivatives of reactive oxygen metabolites (dROMs), which are oxidative stress markers, and C-reactive protein (CRP) in arthritic rats that were exposed to a pressure of 1.25 atmospheres absolute and an oxygen concentration of 36% for 3 weeks (arthritis + HBO group) were compared to those of control rats (control group) and arthritic rats that were not exposed to hyperbaric oxygen (arthritis group). The body weights of the arthritis and arthritis + HBO groups were lower than that of the control group, whereas no difference in the body weight was observed between the arthritis and arthritis + HBO groups. The serum levels of dROMs and CRP in the arthritis group were higher than those in the control and arthritis + HBO groups. No difference in the serum level of CRP was observed between the control and arthritis + HBO groups. These results indicate that the conditions of hyperbaric oxygen exposure used in this study are effective for reducing the levels of reactive oxygen species, which are overproduced during arthritis.


Development and Evaluation of Immunochromatographic Assay for Simple Rapid Detection of Escherichia Coli O157, O26, O111 and Vero Toxin 1/2 in Food Products

International Congress of Meat Science and Technology Proceeding


Chicken collagen hydrolysate protects rats from hypertension and cardiovascular damage.

We previously reported that chicken collagen hydrolysate (CCH) has strong angiotensin I converting enzyme (ACE) inhibitory activity and antihypertensive effects on spontaneously hypertensive rats. Here, we investigated the chronic therapy effects of CCH on blood pressure and vascular relaxation in a cardiovascular damage model of Wistar-Kyoto rats induced by N-nitro-l-arginine methyl ester (L-NAME). Following co-treatment with CCH for 4 weeks, the increment of systolic blood pressure was suppressed significantly. At 8 weeks, the vasorelaxation of thoracic aorta increased significantly, and cardiovascular damage was ameliorated. The concentration of soluble intercellular adhesion molecule-1 (ICAM-1) in blood was reduced significantly by long-term administration of CCH, whereas the nitric oxide concentration was increased significantly at 1 hour post-treatment. The results suggest that beneficial effects of CCH result from antihypertensive function, but also from inhibition of cardiovascular damage to the endothelial cells via its ACE inhibitory activity and regulation of nitric oxide and ICAM-1, which suggests that CCH may be useful as a medicinal food for patients with cardiovascular disease.


ブタプラセンタエキス摂取による色素沈着予防作用

ブタプラセンタエキス摂取による色素沈着予防作用


Chicken collagen hydrolysate reduces proinflammatory cytokine production in C57BL/6.KOR-ApoEshl mice.

Effects of chicken collagen hydrolysate (CCH) on atherogenesis in apolipoprotein E-deficient C57BL/6.KOR-ApoE(shl) mice were investigated in this paper. The mice were fed on either a normal CE-2 diet (control group) or a diet containing 10% CCH (CCH group) for 12 wk. Compared with that of the control, the amount of total plasma cholesterol, total hepatic cholesterol and hepatic triglycerides in the CCH group was reduced by 14.4, 24.7 and 42.8%, respectively. Histological analysis results showed that the abundance of diffuse hepatic lipid droplets and fat vacuoles was decreased in the CCH group. Meanwhile, the concentration of proinflammatory cytokines in the CCH group plasma, including interleukin-6 (IL-6), soluble intercellular adhesion molecule-1 (sICAM-1), and tumor necrosis factor-alpha (TNF-alpha), was downregulated by 43.4, 17.9 and 24.1%, respectively. The present results suggested that CCH treatment might help prevent atherosclerosis through not only its lipid-lowering effects but also inhibiting expression of inflammatory cytokines.

Journal of Nutritional Science and Vitaminology


The bioavailable octapeptide Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro stimulates nitric oxide synthesis in vascular endothelial cells.

Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro (GAXGLXGP, X: Hyp), an octapeptide contained in chicken collagen hydrolysate, inhibits angiotensin I-converting enzyme activity in vitro. Intestinal Caco-2 and bovine aortic endothelial cells (BAECs) were used to investigate whether the transported GAXGLXGP improves vascular function. When GAXGLXGP was added to the apical side of Caco-2 monolayers, the intact form of GAXGLXGP was released to the basolateral side without incorporation into the cells. This transport was energy-independent but was associated with tight junction permeability. GAXGLXGP was then added to BAECs, and endothelial nitric oxide (NO) synthase (eNOS) activation was examined. GAXGLXGP at a concentration of 10 microM stimulated production of NO during a 1 h incubation. This event involved phosphorylation of eNOS at Ser(1179) without a change in the total eNOS protein level. These findings indicate that GAXGLXGP absorbed intact through the intestinal epithelium has direct effects on eNOS activity in vascular endothelial cells, leading to NO synthesis, thereby suggesting the potential for improvement in vascular function.


Molecular size of collagen peptide reverses the permeability of Caco-2 cells.

Intestinal permeability to size-classified fish collagen peptides (FCPs) was examined using Caco-2 monolayers. Minimum-sized FCPs were most efficiently transported across the Caco-2 monolayers. Permeability depended on peptide size. It was independent of the H(+)-coupled peptide transport system but was associated with tight junction permeability, suggesting that the paracellular pathway is responsible for transepithelial transport of collagen peptides.

Bioscience, Biotechnology, and Biochemistry


Differential effects of porcine circovirus type 2 (PCV2) vaccination on PCV2 genotypes at Japanese pig farms.

Infection of pigs with porcine circovirus type 2 (PCV2) causes a variety of disorders collectively referred to as porcine circovirus associated diseases (PCVADs). PCV2 isolates can be classified into two major types: PCV2a and PCV2b. In the present study, effects of vaccination on antibody titers in sera, PCV2 viremia, and shedding of PCV2 in feces were studied on Japanese commercial pig farms where vaccination of piglets against PCV2 was performed using commercially available vaccines. The effectiveness of vaccination against various PCV2 genotypes was also assessed. Among the 16 farms studied, 10 and 6 had been infected with PCV2a and PCV2b, respectively. PCV2a was further subdivided into PCV2a-1 and PCV2a-2. PCV2a-1 and PCV2a-2 prevailed on 6 and 4 farms, respectively, among the 10 farms infected with PCV2a. The PCV2 vaccines were effective in reducing PCV2 infection on commercial pig farms. Mean mortality rates were significantly decreased over 8 months after the start of the PCV2 vaccination program as compared to those before the start of the PCV2 vaccination program on farms infected with PCV2a-2 (20.8% vs. 12.1%) and PCV2b (26.5% vs. 13.7%). On the farms with PCV2a-1 infected pigs, there was no significant difference in the mean mortality rate before versus after the start of the vaccination program (14.7% vs. 14.1%). Mortality rate reduction with the PCV2 vaccination might depend on the genetic types of PCV2.

The Journal of Veterinary Medical Science


Replicated association of the single nucleotide polymorphism in EDG1 with marbling in three general populations of Japanese Black beef cattle.

Marbling, defined by the amount and the distribution of intramuscular fat and measured as beef marbling score (BMS), is an economically important trait of beef cattle in Japan. We recently reported that a single nucleotide polymorphism (SNP), namely, c.-312A>G, in the endothelial differentiation, sphingolipid G-protein-coupled receptor, 1 (EDG1) gene was associated with the BMS level in the Japanese Black beef cattle population of Oita prefecture, with the G allele being associated with a high level of the BMS. Thus, the c.-312A>G SNP seems to be a candidate marker for marker-assisted selection. In this study, we investigated whether this association could be replicated in 3 other independent Japanese Black cattle populations and analyzed the effect of the SNP genotypes on the carcass traits other than the BMS. Statistically significant differences in the BMS level were detected among the genotypes of the c.-312A>G SNP in the Japanese black beef cattle populations of Miyazaki (P = 0.0377) and Nagasaki (P = 0.0012) prefectures, and marginal difference was detected in the Kagoshima prefecture population (P = 0.0786). The G allele in the SNP was associated with an increase in the BMS level.The G allele also seemed to have a favorable influence, if any, on the carcass weight, rib eye area and rib thickness of the cattle populations. These findings suggest that the association of the c.-312A>G SNP with the BMS level in the Japanese Black beef cattle population was replicated in other beef cattle populations, and revealed favorable effects of the G allele on the beef productivity in the general Japanese Black beef cattle population. Thus, we concluded that the c.-312A>G SNP is useful for effective marker-assisted selection to increase the BMS level in Japanese Black beef cattle.


Antithrombotic papain-hydrolyzed peptides isolated from pork meat.

Atherothrombotic disease such as coronary artery disease and stroke is one of major causes of death. Platelets play important role in these arterial diseases. Suppression of platelet activity by materials may decrease incidence of the disease. The present study aimed to examine the peptides from pork meat with antithrombotic activity. Peptide fraction or hydrolyzate of defatted pork meat by papain was used as the starting material. Antithrombotic activity of the fraction was assessed by a share-induced platelet function test in vitro (haemostatometry) using non-anticoagulated rat blood, followed by a helium-neon laser-induced mouse carotid artery thrombosis test in vivo. The starting peptide fraction with mean molecular weight 2500 showed antithrombotic activity in vivo after oral administration to mice at 210 mg/kg body weight. The fraction with mean molecular weight 2517 further purified by cation exchange chromatography showed antithrombotic activity after oral administration at 70 mg/kg body weight. Antithrombotic activity of the purified peptide fraction was equivalent to that of aspirin at 50 mg/kg body weight. It is possible this pork peptide could be beneficial to prevent atherothrombosis.


Porcine artery elastin preparation reduces serum cholesterol level in rats


鶏コラーゲン加水分解物(C-COP)の血管保全作用

鶏コラーゲン加水分解物(C-COP)の血管保全作用


Blood Concentration of Food-Derived Peptides following Oral Intake of Chicken Collagen Hydrolysate and its Angiotensin-Converting Enzyme Inhibitory Activity in Healthy Volunteers

鶏コラーゲン加水分解物摂取後のヒト血中ペプチドの動態とACE阻害作用

In this study, we investigated the concentration of food derived-peptides in blood following oral intake of chicken collagen hydrolysate (chicken-collagen octa peptides ; C-COP) and measured the angiotensin-converting enzyme (ACE) inhibitory activity of these peptides. Healthy volunteers ingested the C-COP solution (25g/60kg body weight) after fasting for 12h. The peptide-formed hydroxyproline (Hyp) significantly increased and reached maximum levels (91.1μM) 2h after consumption. Pro-Hyp, Leu-Hyp, Pro-Hyp-Gly, Phe-Gly, Ala-Hyp, Hyp-Gly, Glu-Hyp-Gly, Ala-Hyp-Gly and Ser-Hyp-Gly were detected in human blood, possessing ACE-inhibitory activity. Of them, Ala-Hyp (IC50 : 0.177mM) had the strongest ACE inhibitory activity. Thus, C-COP appears to absorb with the peptides by enzymatic digestion in blood, thereby inhibiting ACE, suggesting that food-derived collagen peptides are related to antihypertensive effects.

降圧作用を有する鶏コラーゲン加水分解物(C-COP)を健常人に摂取させ，C-COPの血中動態およびアンジオテンシン変換酵素(ACE)阻害活性との関連性について検討した．
体重60kgあたり25gのC-COPをヒトに単回摂取させた場合，血中に遊離ヒドロキプロリン(Hyp)とともに，ペプチド型のHypも確認された．血中のペプチド型Hyp量は，2時間後(91.1μmol/L)に最大となった．摂取2時間後の血中からPro-Hyp, Leu-Hyp, Pro-Hyp-Gly, Phe-Hyp, Ala-Hyp, Hyp-Gly, Glu-Hyp-Gly, Ala-Hyp-Gly, Ser-Hyp-GlyのHyp含むペプチドが確認された．血中ペプチドは，ACEに対する阻害活性を有しており，Ala-Hyp(IC50 : 0.177mmol/L)の活性がもっとも高かった．以上より，C-COPの降圧作用は，血中ペプチドによるACE阻害が作用機序であると考えられた．


Effects of liquid whey supplement on the productivity of pigs in a commercial pig farm

養豚農場における液状ホエーの給与がブタの生産性へ与える影響

The productivity of pigs given commercial formula feed supplemented with liquid whey, which was discharged from a dairy factory, was examined. A total of 66 weaned piglets from 9 litters were divided into the group of pigs given commercial formula feed supplemented with liquid whey (Whey) and the group given commercial formula feed alone (CN), considering possible hereditary interference. A total of 267 L of liquid whey was given per animal. Body weight was measured periodically. The blood sample was taken to compare innate immune function between the Whey group and CN group immediately before shipment of pigs. Mean body weight at ages of 65 days and 173 days was significantly higher in the Whey group than in the CN group (P < 0.05). Daily weight gain was significantly higher in the Whey group at ages from 29 to 65 days (P < 0.01) and from 105 to 173 days (P < 0.05), as compared with the control group. There were no significant differences in carcass traits of meat quality or innate immune function between the Whey group and the CN group. These results showed that liquid whey supplement allows acceleration of age at slaughter without affecting carcass traits or meat quality and is useful in improving the productivity of pigs.

乳業工場から排出された液状ホエーを通常飼料に付加的に給与し，豚肉生産試験を行った．9腹の離乳豚66頭を通常飼料のみで飼育した対照区(CN区)と通常飼料に液状ホエーを付加的に給与したホエー区(Whey区)に母豚効果を配慮して分けた．Whey区には試験期間を通して267 L/頭の液状ホエーを給与した．両区は経時的に体重測定を行い，出荷直前に採血をして自然免疫の状態を比較した．平均体重は65日齢と173日齢でWhey区の方が有意に高かった(P < 0.05)．日増体量はWhey区の方が生後29-65日の期間(P < 0.01)と105-173日の期間(P < 0.05)で有意に高かった．また，枝肉成績およびロース肉の理化学的特性はWhey区とCN区とで有意差は認められず，自然免疫状態についても有意差がなかった．以上の結果から，液状ホエー給与は枝肉成績および肉質に影響を与えることなくブタの出荷日齢を短縮する傾向にあり，生産性向上に有効であることが示された．


Association of a single nucleotide polymorphism in ribosomal protein L27a gene with marbling in Japanese Black beef cattle.

Marbling, defined by the amount and distribution of intramuscular fat, is an economically important trait of beef cattle in Japan. The c2-11#2 expressed sequence tag (EST) has been previously shown to possess expression difference in musculus longissimus muscle between low-marbled and high-marbled steer groups, and to be located within genomic region of a quantitative trait locus for marbling. Thus, the ribosomal protein L27a (RPL27A) gene containing the c2-11#2 EST sequence was considered as a positional candidate for the gene responsible for marbling. In the present study, a single nucleotide polymorphism (SNP) in the promoter region of the RPL27A, referred to as g.3109537C>T, was detected between the 2 steer groups. The SNP was associated with the predicted breeding value for beef marbling standard number by the analyses using Japanese Black beef cattle population. The effect of genotypes of the SNP on the predicted breeding value for subcutaneous fat thickness was not statistically significant. These findings suggest that the RPL27A SNP may be useful for effective marker-assisted selection to increase the levels of marbling in Japanese Black beef cattle.


Novel SNP in 5' flanking region of EDG1 associated with marbling in Japanese Black beef cattle.

Marbling, defined by the amount and distribution of intramuscular fat, is an economically important trait of beef cattle in Japan. The endothelial differentiation, sphingolipid G-protein-coupled receptor, 1 (EDG1) gene has been considered as a positional functional candidate for the gene responsible for marbling. We have recently reported that 2 single nucleotide polymorphisms (SNPs), c.-312A>G in the 5' untranslated region (UTR) and c.*446G>A in the 3' UTR in EDG1 were associated with marbling in Japanese Black beef cattle, but this was not functional and a causal mutation for marbling. In the present study, we detected 2 novel SNPs, referred to as g.1475435G>A and g.1471620G>T, in the 5' flanking region of the EDG1 between low-marbled and high-marbled steer groups, which were previously shown to have EDG1 expression differences in musculus longissimus muscle. The g.1475435G>A SNP seemed not to segregate in Japanese Black beef cattle. The g.1471620G>T SNP was associated with the predicted breeding value for beef marbling standard number by the analyses using Japanese Black beef cattle population. Based on these findings, we hypothesized that the g.1471620G>T SNP might have an impact on EDG1 expression and also marbling.


Efficacy of Lactobacillus plantarum strain HSK201 in relief from Japanese cedar pollinosis.

To determine the effects of Lactobacillus plantarum strain HSK201 on Japanese cedar pollinosis, a single-blind, placebo-controlled study was conducted in 2008. The HSK201 group was administered fermented milk prepared with HSK201 for 8 weeks, and the placebo group was administered non-fermented milk adjusted to the same acidity and taste. We found HSK201 strain intake to suppress both helper T cell type 1/2 ratio reduction and serum Japanese cedar pollen-specific IgE elevation at the peak of pollen dispersion. In addition, the nasal and ocular symptom scores in the HSK201 group were also lower than those in the placebo group during the early phase of the pollen season. Although this was a preliminary study with 19 employees of our own company serving as subjects, the results suggest that ingestion of the HSK201 strain alleviates pollinosis symptoms during the period when pollen exposure is low and the symptoms are mild.

Bioscience, Biotechnology, and Biochemistry


Ingestion of ELASTIN PEPTIDE Alters MessengerRNA expression in Mice Skin

Current Topics in Nutraceutical Research


ブタ由来エラスチンペプチドの経口摂取によるマウス皮膚水分量の向上

ブタ由来エラスチンペプチドの経口摂取によるマウス皮膚水分量の向上


Oral Administration of chicken Breast Extract Activates Serotonin Metabolism in the Hippocampus of Rats

Journal of Animal and Veterinary Advances


Chicken breast attenuates high-intensity-exercise-induced decrease in rat sarcoplasmic reticulum Ca2+ handling.

This study was conducted to determine whether dietary chicken-breast extract (CBEX), a rich source of histidine-containing dipeptides, could modify exercise-induced changes in sarcoplasmic reticulum (SR) function. After 5 weeks of dietary CBEX, SR Ca2+-handling ability was examined in the vastus lateralis muscles of rats subjected to high-intensity running for 2.5 min. Dietary CBEX caused an approximately 15% and 45% increase (p<.01) in muscle carnosine and anserine concentrations, respectively. In resting muscles, depressions in SR Ca2+-ATPase activity were evoked by dietary CBEX without concomitant changes in SR Ca2+ uptake and release rates. The data confirm that high-intensity exercise depresses SR Ca2+ handling. In spite of the same run time, SR Ca2+ handling was reduced to a lesser degree in muscles of CBEX-containing-chow-fed rats than in standard-chow-fed rats (p<.05). These results suggest that dietary CBEX might attenuate deteriorations in SR Ca2+-handling ability that occur with high-intensity exercise.

International Journal of Sport Nutrition and Exercise Metabolism


Lactobacillus rhamnosus GG and Lactobacillus casei suppress Escherichia coli-induced chemokine expression in intestinal epithelial cells.

Recently, some strains of lactic acid bacteria (LAB) have been reported to prevent the development of atopic dermatitis and to improve allergic symptoms, especially in young children. However, the mechanisms involved in these effects are not fully understood. Intestinal microbiota play critical roles in the development of host immune development and are recognized and regulated by the host through intestinal epithelial cells (IECs). We thus hypothesized that LAB influence the host immune system through the activation of IECs. To begin testing this hypothesis, chemokine expression in IECs exposed to intestinal bacteria was investigated. Caco-2 cell monolayers were stimulated with different concentrations of various live or heat-killed intestinal bacteria or bacterial components for up to 3 h. Changes in the gene expressions of various chemokines were measured using quantitative real-time PCR. The expressions of CCL20, CXCL8, CXCL10 and CX3CL1 were strongly induced by nonpathogenic Escherichia coli in a dose-dependent manner and were partially induced by some commensal LAB. In contrast, Lactobacillus rhamnosus GG (LGG) and Lactobacillus casei did not induce these chemokine expressions. In addition, LGG significantly suppressed the expressions of CCL20 and CXCL10 induced by E. coli, peptidoglycan or flagellin when cultured simultaneously. LGG and L. casei markedly suppressed E. coli-induced chemokine expression, presumably through the suppression of the Toll-like receptor-mediated signal transduction pathway, at least in part. The clinical importance of this suppressive effect and the mechanisms involved require further investigation; however, such effects can be used as a marker to identify clinically useful LAB.

International Archives of Allergy and Immunology


Royal jelly peptides inhibit lipid peroxidation in vitro and in vivo.

Royal jelly peptides (RJPx) isolated from hydrolysates of water-soluble royal jelly proteins prepared with protease P exhibited significantly stronger hydroxyl radical-scavenging activity (p<0.001), and antioxidant activity against lipid peroxidation (LPO, p<0.001), than did water-soluble royal jelly protein (WSRJP) in vitro. We also investigated the in vivo antioxidant activity of RJPx against ferric nitrilotriacetate (Fe-NTA)-induced LPO. Male Wistar rats were divided into a control group (Group C), an Fe-NTA group (Group Fe), and an Fe-NTA with RJPx group (Group Fe+R). Rats in Group Fe+R were fed RJPx (2 g/kg body weight) daily for 5 wk. Fe-NTA (8 mg Fe/kg body weight) was then intraperitoneally injected, and serum lipid levels were examined 2 h later. Serum total cholesterol (TC) levels were lower (p<0.05) while low-density lipoprotein (LDL) and LPO were significantly higher (p<0.01) in Group Fe than in Group C. TC (p<0.05) and LPO levels (p<0.01) were lower in Group Fe+R than in Group Fe. Our data suggest that RJPx may inhibit LPO both in vitro and in vivo.

Journal of Nutritional Science and Vitaminology


Genotypic change of porcine circovirus type 2 on Japanese pig farms as revealed by restriction fragment length polymorphism analysis.

Porcine circovirus type 2 (PCV2) has been recognized as the causal agent of postweaning multisystemic wasting syndrome and can be divided into two major genotypic groups. We developed a method of restriction fragment length polymorphism (RFLP) analysis of PCV2 open reading frame 2 for easy discrimination between the two major groups. Genotyping of PCV2 isolates from 10 Japanese commercial pig farms was performed, and the analysis revealed that both PCV2 groups and at least five RFLP types of PCV2 are prevalent in Japan. On two farms, the genotypes of the PCV2 isolates in the spring of 2007 were different from those in the autumn of 2006. One genotype may have become dominant within only six months on these farms.

The Journal of Veterinary Medical Science


鶏コラーゲン加水分解物(C-COP)を配合した乳酸菌飲料による正常高値血圧者に対する降圧作用のサブグループ解析


鶏コラーゲン加水分解物(C-COP)を配合した乳酸菌飲料の過剰摂取時における安全性


鶏コラーゲン加水分解物(C-COP)を配合した乳酸菌飲料による軽症高血圧者および正常高値血圧者に対する降圧作用と安全性の検討


Specific discrimination of chicken DNA from other poultry DNA in processed foods using the polymerase chain reaction.

In the present study, specific discrimination of chicken DNA contamination in processed foods using the polymerase chain reaction was investigated. The primer pair was designed to amplify a 102-bp fragment of the chicken mitochondrial 16S ribosomal RNA gene. While the DNA from chicken meat was amplified, the DNA from other poultry meat, mammalian meat, fish, shellfish, and cereals was not amplified. The primer amplified DNA fragments derived from model processed and nonprocessed food samples containing 0.001, 0.01, 0.1, 1, 10, and 100% chicken.

Bioscience, Biotechnology, and Biochemistry


mRNA expression of lysyl oxidase and matrix metalloproteinase-12 in mouse skin.

Elastic fibers in the dermis play an important role in skin elasticity. The desmosine crosslinking structure constructed of lysyl oxidase (LOX) in elastic fibers contributes to elasticity, while elastic fibers are primarily degraded by one of the matrix metalloproteinases (MMPs), MMP-12. We investigated the gender differences and diurnal variation of these enzymes. Gender-based differences in LOX mRNA expression were detected, and were significantly lower in females. In contrast, higher MMP-12 mRNA expression was observed in the light period, suggesting that elastic fibers might be degraded in the light rather than the dark period.


Carnosine-induced antidepressant-like activity in rats.

Depression is a pathological state of mood and is considered as one of the major causes of disabilities. Thus, the prevention of depression and care for individuals with depression is important. In the present study, we examined whether a single oral dose of CBEX (chicken breast extract), or carnosine (one of the major components of CBEX) affects immobility time, an index of depressive-like behavior, in the forced swimming test in male Wistar rats. CBEX tended to (P=0.09) and carnosine significantly (P<0.05) decreased immobility time in the forced swimming test. In the hippocampus, both CBEX and carnosine significantly decreased 3-methoxy-4-hydroxyphenylglycol, a major metabolite of norepinephrine, indicating that CBEX and carnosine could reduce NE activity in the hippocampus in the forced swimming test. CBEX and carnosine did not affect total locomotive distance or rearing in the open field test, suggesting that the reductions of immobility time by both treatments in the forced swimming test were not merely due to the stimulation of general motor activity. Taken together, these results suggest that CBEX has an antidepressant-like effect, which may be due, in part, to the effect of carnosine.

Pharmacology, Biochemistry, and Behavior


Hypotensive Action of Chicken Collagen Hydrolysate in Lactic Acid Beverage on Mildly Hypertensive Subjects

鶏コラーゲン加水分解物(C-COP)を配合した乳酸菌飲料の軽症高血圧者に対する降圧作用の検討

Journal of Japanese Society of Clinical Nutrition


An Angiotensin I-converting Enzyme (ACE)-inhibitory Peptide Derived from Chicken Collagen Hydrolysate Lowers Blood Pressure in Spontaneously Hypertensive Rats

高血圧自然発症ラットにおける鶏コラーゲン由来オクタペプチドの血圧降下作用

We studied the antihypertensive effect of an octapeptide (Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro), which is derived from chicken collagen hydrolysates and has strong angiotensin I-converting enzyme (ACE)-inhibitory activity, on spontaneously hypertensive rats (SHRs). 6h after a single oral administration of the octapeptide (4.5mg/kg SHR body weight), the systolic blood pressure was significantly reduced. The octapeptide was not digested by digestive enzymes from the epithelia of the porcine small intestines <I>in vitro.</I> These results suggest that this ACE-inhibitory peptide is absorbed by SHRs in the peptide form and that it lowers blood pressure.

本研究では，鶏コラーゲンペプチド中に含まれ，強いACE阻害活性を有するオクタペプチドのSHRへの血圧降下作用および消化酵素による分解性について検討した．
(1) オクタペプチドをSHRに4.5mg/kg体重で単回投与した結果，SHRの収縮期血圧値を有意に低下させた．
(2) 試験管レベルでオクタペプチドは，ペプシンおよび，トリプシン/キモトリプシン，小腸上皮酵素に対して難消化性を示し，そのACE阻害活性は維持されていた．
以上の結果から，鶏コラーゲンペプチドに含まれるオクタペプチドは，in vitroでのACE阻害活性を持つだけでなく，SHRの血圧降下作用を示すことが確認された．


Angiotensin I-converting enzyme-inhibitory peptides obtained from chicken collagen hydrolysate.

In this study, collagen extracted from chicken legs (which are the yellow keratin parts containing a nail) was hydrolyzed with various enzymes, and the angiotensin I-converting enzyme (ACE)-inhibitory activity of each hydrolysate was determined. The hydrolysate by treatment with an Aspergillus species-derived enzyme had the highest activity (IC 50 = 260 microg/mL). The fraction of this hydrolysate obtained by ultrafiltration with a molecular-weight cutoff of 3000 Da (low fraction) had a stronger activity (IC 50 = 130 microg/mL) than the fractionated one. This fraction was further fractionated by HPLC, and the peptides in the fraction with high ACE-inhibitory activity were identified. The amino acid sequences of the four peptides were identified using a protein sequencer. These peptides were synthesized to confirm their ACE-inhibitory activities; this showed that peptides with a Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro sequence had the highest activity (IC 50 = 29 microM). When the low fraction was administered to spontaneous hypertensive rats, a decrease in their blood pressure was observed after 2 h of administration, and a significant decrease in blood pressure (-50 mmHg) was observed after 6 h. Moreover, long-term administration studies indicated that the low fraction showed a significant suppression of increased blood pressure.

Journal of Agricultural and Food Chemistry


Bone density of the femur and fiber cross-sectional area and oxidative enzyme activity of the tibialis anterior muscle in type II collagen-induced arthritic mice.

Femur bone densities and tibialis anterior muscle properties of type II collagen-induced arthritic mice were determined. Furthermore, voluntary running activities of arthritic mice were compared with those of controls. Arthritis was induced by an intradermal injection of type II collagen in the adjuvant. Body and muscle weights were lower in arthritic mice than in controls. Cortical and trabecular bone densities and muscle fiber cross-sectional areas were decreased by arthritis. After classifying the arthritic severity into slight, intermediate, and severe levels based on the degree of knuckle swelling, cortical and trabecular bone densities, fiber cross-sectional areas, and fiber succinate dehydrogenase activities were lowest when arthritis was most severe. Furthermore, arthritic mice, especially those with intermediate and severe levels, showed lower voluntary running activities. These findings indicate that lower bone density and muscle atrophy of type II collagen-induced arthritic mice are related to arthritic severity and decreased motor activity.


Polymorphisms in promoter regions of PDHB, SORBS1, and EDG1 genes showing marbling-associated expression changes.

We have recently showed that the pyruvate dehydrogenase (lipoamide) beta (PDHB) gene involved in fatty acid oxidation, the sorbin and SH3 domain containing 1 (SORBS1) gene involved in insulin-stimulated glucose uptake, and the endothelial differentiation, sphingolipid G-protein-coupled receptor, 1 (EDG1) gene involved in blood vessel formation possess expression differences in musculus longissimus muscle between low-marbled and high-marbled steer groups. In the present study, we detected single nucleotide polymorphisms (SNPs) in the promoter regions of the 3 genes between the 2 steer groups. A SNP in the EDG1 exhibited significantly different allelic distribution between animals with extremely high predicted breeding value for marbling and with extremely low one. The EDG1 SNP may be related to changes in gene expression and/or marbling.

Research Communications in Molecular Pathology and Pharmacology


Safety evaluation of chicken breast extract containing carnosine and anserine.

Chicken breast extract (CBEX) is obtained via hot water extraction of chicken breast and contains among its primary constituents carnosine and anserine, which are histidine-containing dipeptides present in the muscle tissues of most vertebrate species. Dietary intake of CBEX has been previously shown to buffer hydrogen ions formed during high-intensity exercise in human skeletal muscle cells, thereby inhibiting a decrease in muscle cell pH and subsequent muscle fatigue. The objective of this paper is to report the results of safety studies completed on CBEX. CBEX was determined to have an oral LD(50) value of more than 6000 mg/kg body weight in rats. Gavage doses of 500 or 2000 mg CBEX/kg body weight/day administered to rats for 90 days produced no toxicologically significant, dose-related, differences between control and treated animals with respect to body weight gain, food consumption, behavioral effects, hematological and clinical chemistry parameters, absolute and relative organ weights, or gross and microscopic findings. In the presence or absence of metabolic activation, CBEX exerted no mutagenic activity in the Ames assay conducted in various strains of Salmonella typhimurium and Escherichia coli. The results of these studies support the safety of CBEX as a potential dietary source of carnosine and anserine.


Oral administration of chicken breast extract increases brain carnosine and anserine concentrations in rats.

Carnosine (beta-alanyl-L-histidine) and its derivative anserine (beta-alanyl-1-methyl-L-histidine) are antioxidants and putative neurotransmitters in the brain. These dipeptides are rich in the commercially available supplement chicken breast extract (CBEX). To clarify the effects of CBEX on the brain, we examined whether single oral administration of CBEX (20 ml/kg) affects brain dipeptide and free amino acid concentrations in male Wistar rats. CBEX significantly and time-dependently increased carnosine and anserine levels in the plasma (at 120 min after injection, increase rates were 2976 and 4142%, respectively), hippocampus (64 and 78%), and hypothalamus (188 and 120%), but not in cerebral cortex. Significant and time-dependent increases in citrulline in the hippocampus (49%) and hypothalamus (41%) demonstrated generation of nitric oxide due to the increased carnosine and/or anserine levels in these brain regions. These findings suggest that CBEX modifies brain functions by increasing levels of these dipeptides.


Polymorphisms for 7 expressed sequence tags showing marbling-associated expression changes.

We have recently showed that the 7 expressed sequence tags (ESTs), c2-11#2, c3-3, c20-29, c22-3, c26-18#1, c26-42, and g5-10, possess expression differences in musculus longissimus muscle between low-marbled and high-marbled steer groups. In the present study, we detected single nucleotide polymorphisms (SNPs) in the 5' flanking regions of the 7 EST sequences between the 2 steer groups. A SNP for the c2-11#2 EST exhibited significantly different allelic distribution between animals with extremely high predicted breeding value for marbling and with extremely low one. The SNP in the ribosomal protein L27a (RPL27A) gene containing the c2-11#2 EST sequence may be related to changes in gene expression and/or marbling.


Effects of hyperbaric exposure with high oxygen concentration on the physical activity of developing rats.

The effects of hyperbaric exposure with high oxygen concentration on the physical activity of developing male rats were investigated. Five-week-old male rats were exposed to an atmospheric pressure of 1.25 with an oxygen concentration of 36.0% for 12 h (7.00-19.00 h) and exercised voluntarily for 12 h (19.00-7.00 h) daily for 8 weeks. The voluntary running activities were compared with those in age-matched rats without hyperbaric exposure. In addition, the properties of the soleus and plantaris muscle fibers and their spinal motoneurons were examined. The voluntary running activities of rats with or without hyperbaric exposure increased during development. However, the mean voluntary running activities were higher in rats with hyperbaric exposure (7,104 m/day) than in those without hyperbaric exposure (4,932 m/day). The oxidative capacities of the soleus and plantaris muscle fibers and their spinal motoneurons increased following hyperbaric exposure. It is suggested that adaptations of neuromuscular units to hyperbaric exposure with high oxygen concentration enhance the metabolism, and thus, the function of neuromuscular units is promoted.


Royal jelly supplementation improves lipoprotein metabolism in humans.

Royal jelly (RJ) has several physiological effects and is widely used in commercial medical products and health foods. We examined the effects of RJ supplementation on serum lipoprotein metabolism in humans. Fifteen volunteers were divided into an RJ intake group (n=7) and a control group (n=8). The RJ group took 6 g per day for 4 wk. Their serum total cholesterol (TC) and serum low-density lipoprotein (LDL) decreased significantly compared with those of the control group (p<0.05). There were no significant differences in serum high-density lipoprotein (HDL) or triglyceride concentrations. Moreover, the relationship between the serum cholesterol and lipoprotein levels was investigated. Among the lipoprotein fractions, small very-low-density lipoprotein was decreased (p<0.05) after RJ intake. Our results suggest that dietary RJ decreases TC and LDL by lowering small VLDL levels.

Journal of Nutritional Science and Vitaminology


A judgment on postmortem aging in longissimus dorsi based on a peptide substrate library.

We attempted to develop a method to determine easily and effectively the degree of postmortem aging of pork longissimus dorsi (LD) by measuring the activity of proteases in the LD using fluorogenic peptide substrates. LD was used to measure the change with time in the protease activity detected with these substrates. Determining the variations within the LD muscles, strong positive correlations were found between changes in hardness and fluorescence intensities against Ac-Ala-MCA, Ac-Met-MCA, Ac-Ser-MCA, Ac-Thr-MCA, and Ac-Ala-Phe-MCA (P<0.005), and strong negative correlations were found between changes in total amounts of free amino acids and Ac-Ala-MCA, Ac-Met-MCA, Ac-Ser-MCA, Ac-Thr-MCA, and Ac-Ala-Phe-MCA (P<0.001). Negative correlations were also observed between changes in the amounts of free Ala, Arg, Lys, Leu, Met, Phe, and Tyr and the fluorescence intensities against Ala, Arg, Lys, Leu, Met, Phe, and Tyr-MCA respectively (P<0.001).

Bioscience, Biotechnology, and Biochemistry


Action mechanism of an angiotensin I-converting enzyme inhibitory peptide derived from chicken breast muscle.

In a previous study, we isolated the inhibitory peptide (P4 peptide, Gly-Phe-Hyp-Gly-Thr-Hyp-Gly-Leu-Hyp-Gly-Phe) for angiotensin I-converting enzyme (ACE) from chicken breast muscle extract possessing hypotensive activity for spontaneously hypertensive rats (SHRs). This study was performed to elucidate the peptide's action mechanisms of inhibiting ACE. Intravenous administration of synthetic P4 peptide resulted in significant drops in the blood pressures of SHRs. As Dixon plots indicate, the P4 peptide showed high affinity toward ACE (K(i) = 11.48 microM) and only 10% of the total amount of the P4 peptide was decomposed. The analyses of the relationship between the ACE inhibitory activity and structure of the P4 peptide clarified that Hyp-Gly-Leu-Hyp-Gly-Phe showed a stronger activity (IC50 = 10 microM) than the P4 peptide (IC50 = 46 microM). When Phe at the C-terminus of the P4 peptide was deleted, IC50 changed to 25000 microM, indicating that Phe at the C-terminus of the peptide is very important for ACE inhibitory activity.

Journal of Agricultural and Food Chemistry


Consumption of pork-liver protein hydrolysate reduces body fat in Otsuka Long-Evans Tokushima Fatty rats by suppressing hepatic lipogenesis.

This study was performed to examine the effect of consumption of pork-liver protein hydrolysate (PLH) on body fat accumulation in Otsuka Long-Evans Tokushima Fatty (OLETF) rats as a non-insulin-dependent diabetes mellitus model and in Long-Evans Tokushima Otsuka (LETO) rats as a control. Male 20-week-old OLETF and LETO rats were pair-fed either PLH or casein containing diet for 14 weeks. In the OLETF rats, dietary PLH significantly reduced the growth and weight of fat pad including perirenal and epididymal adipose tissues. Consumption of PLH markedly suppressed hepatic activities of lipogenesis enzymes such as glucose-6-phosphate dehydrogenase and fatty acid synthase and slightly elevated fecal excretion of total fat. In the LETO rats, growth and adipose tissue weight were unaffected by dietary treatment. The results suggest that PLH is a novel ingredient suppressing body fat in genetically obese rats by reducing lipogenesis.


調理加工モデル食品を用いたアレルゲン検査用イムノクロマトキットの評価

調理加工モデル食品を用いたアレルゲン検査用イムノクロマトキットの評価


Effect of dietary vegetable and animal proteins on atherothrombosis in mice.

It is believed that vegetable and fish, but not animal, proteins prevent thrombosis. The present study compared the effect of long-term intake of purified vegetable and animal proteins (casein, pork, egg white, chicken, white and red fish, soybean, and potato) and powders from whole vegetable and animal meats (soybean, pork, chicken, and horse mackerel) on thrombotic tendency. Western-style high-fat diets with various proteins were given to mice deficient in apolipoprotein-E and low-density lipoprotein receptor for 8 or 12 wk. Thrombotic tendency was assessed by a laser-induced thrombosis test, severity of atherosclerosis was assessed by morphometric analysis of the aortic lesions, and reactivity of platelets was measured by a shear-induced platelet function test. There was no difference in thrombotic tendency between the vegetable and animal protein diets with the exception of pork protein, which showed a prothrombotic effect. The latter was not observed in animals kept on a high-protein diet. Thrombotic tendency was not affected by preparation of dietary powders from whole pork, chicken, or horse mackerel meats and casein (control), all of which contained the same percentages of protein and lipid. In contrast, soybean powder showed an antithrombotic effect, which was mainly due to an inhibition of atherogenesis rather than to platelet inhibition. Neither the purified animal nor the vegetable protein diet affected thrombotic tendency. Soybean powder inhibited atherothrombosis mainly by inhibiting the atherosclerotic process.


Carnosine and anserine ingestion enhances contribution of nonbicarbonate buffering.

The purpose of the present study was to investigate the effect of supplementation with chicken breast extract (CBEX), which was a rich source of carnosine and anserine, on acid-base balance and performance during intense intermittent exercise. Eight male subjects performed intense intermittent exercise that consisted of 10 x 5-s maximal cycle ergometer sprints with a 25-s recovery period between each sprint. The subjects ingested 190 g of the test soup containing either CBEX or a placebo 30 min before the commencement of exercise. Arterial blood samples were collected at rest and during exercise to estimate the carnosine and anserine concentrations, pH, and bicarbonate concentration ([HCO3-]). Concentrations of anserine and its related amino acid significantly increased 30 min after CBEX supplementation, as compared with their values at rest. However, carnosine did not increase significantly. Following CBEX supplementation, the pH was significantly higher (P < 0.05) at the end of exercise, and [HCO3-] was also significantly higher (P < 0.05) during the latter half of exercise and after exercise. There were no significant differences in the total power and mean power of each set between the CBEX and placebo supplemented groups. Although oral supplementation with CBEX (which is a rich source of carnosine and anserine) increased the contribution of the nonbicarbonate buffering action and decreased bicarbonate buffering action in blood, intense intermittent exercise performance did not improve significantly.

Medicine and Science in Sports and Exercise


Simultaneous determination of residual veterinary drugs in bovine, porcine, and chicken muscle using liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

A simple and rapid method using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC/MS/MS) for the simultaneous determination of 130 veterinary drugs and their metabolites in bovine, porcine, and chicken muscle was developed. The drugs (1 to 10 ng/g, in muscle) were extracted from bovine, porcine, or chicken muscles with acetonitrile-methanol (95:5, v/v), and the extracts were delipidated with n-hexane saturated with acetonitrile. The extracts were evaporated, dissolved with methanol, analyzed by liquid chromatography with gradient elution on a C18 column, and determined by electrospray ionization tandem mass spectrometry. The detection limits ranged from 0.03 to 3 ng/g. The quantitation limits ranged from 0.1 to 10 ng/g. One hundred eleven, 122, and 123 drugs from bovine, porcine, and chicken muscle respectively showed recoveries between 70 and 110%.


Effects of Carnosine and Anserine Supplementation on Relatively High Intensity Endurance Performance

International Journal of Sport and Health Science


Identification of food-derived collagen peptides in human blood after oral ingestion of gelatin hydrolysates.

In the present study, we identified several food-derived collagen peptides in human blood after oral ingestion of some gelatin hydrolysates. Healthy human volunteers ingested the gelatin hydrolysates (9.4-23 g) from porcine skin, chicken feet, and cartilage after 12 h of fasting. Negligible amounts of the peptide form of hydroxyproline (Hyp) were observed in human blood before the ingestion. After the oral ingestion, the peptide form of Hyp significantly increased and reached a maximum level (20-60 nmol/mL of plasma) after 1-2 h and then decreased to half of the maximum level at 4 h after the ingestion. Major constituents of food-derived collagen peptides in human serum and plasma were identified as Pro-Hyp. In addition, small but significant amounts of Ala-Hyp, Ala-Hyp-Gly, Pro-Hyp-Gly, Leu-Hyp, Ile-Hyp, and Phe-Hyp were contained.


筋肉中に残留する抗生物質および合成抗菌剤のLC/MS/MSを用いた同時分析

筋肉中に残留する抗生物質および合成抗菌剤のLC/MS/MSを用いた同時分析


LC/MS/MSを用いた筋肉中の残留動物用医薬品の多成分同時分析

LC/MS/MSを用いた筋肉中の残留動物用医薬品の多成分同時分析


Hyperbaric exposure with high oxygen concentration enhances oxidative capacity of neuromuscular units.

The effects of hyperbaric exposure with high oxygen concentration on spinal motoneurons and the skeletal muscle fibers that they innervate were investigated. Five-week-old male rats were exposed to a hyperbaric (1.25 atmospheric pressure) environment with a high oxygen concentration (35.0%) for 6h daily. The number, cell body size, and oxidative enzyme activity of motoneurons innervating the soleus and plantaris muscles were examined after 8 weeks of hyperbaric exposure. In addition, the fiber type distribution, cell size, and oxidative enzyme activity of the slow soleus and fast plantaris muscles were examined. The oxidative enzyme activity of alpha motoneurons innervating the soleus and plantaris muscles increased after hyperbaric exposure, irrespective of their cell body sizes. The percentage of high-oxidative fibers in the soleus and plantaris muscles increased after hyperbaric exposure. The oxidative enzyme activity of all types of fibers in the soleus and plantaris muscles increased after hyperbaric exposure. It is concluded that hyperbaric exposure with high oxygen concentration enhances the oxidative capacity of neuromuscular units.


特定原材料(落花生)測定の厚生労働省通知ELISA法確立のための複数機関による評価研究


特定原材料(そば)測定の厚生労働省通知ELISA法確立のための複数機関による評価研究


トリ胸肉抽出物(CBEXTM)の経口摂取が高強度間欠的運動パフォーマンスに及ぼす影響

Japan Journal of Physical Education, Health and Sport Sciences


特定原材料(小麦)測定の厚生労働省通知ELISA法の複数機関による評価研究

特定原材料(小麦)測定の厚生労働省通知ELISA法の複数機関による評価研究


特定原材料(牛乳)測定の厚生労働省通知ELISA法の複数機関による評価研究

特定原材料(牛乳)測定の厚生労働省通知ELISA法の複数機関による評価研究


トリ胸肉抽出物(CBEXTM)長期摂取が骨格筋中カルノシン濃度と短時間高強度運動パフォーマンスに及ぼす影響

Japanese Journal of Physical Fitness and Sports Medicine


特定原材料(卵)測定の厚生労働省通知ELISA法の複数機関による評価研究

特定原材料(卵)測定の厚生労働省通知ELISA法の複数機関による評価研究


Synergistic Effects of D-Glucosamine and Collagen Peptide on Healing Experimental Cartilage Injury


Some human B and T cell epitopes of bovine serum albumin, the major beef allergen

Bovine serum albumin (BSA) is the major beef allergen. Since IgE and T cell recognitions are central to the specific immune response to allergens, the identification and immunologic characterization of B and T cell epitopes of BSA represent important steps in the development of treatments for beef allergy. Prior to our experiments, we hypothesized that BSA-specific antibodies and T cells react primarily with sequential epitopes in which the amino acid sequences differ greatly between bovine and human albumin. To clarify this hypothesis, 16 peptides corresponding to such regions were synthesized as candidate epitopes. Among them, at least two regions, aa336-345 and aa451-459, were found to be B cell (IgE-binding) epitopes. In inhibition ELISA experiments, EYAV (aa338-341) and LILNR (aa453-457) bound to patient IgE antibodies and were found to be the cores of the IgE-binding epitopes. Three regions, DDSPDLPKLKPDPNTLC (aa107-123), PHACYTSVFDKLKHLVDEP (aa364-382), and LSLILNRLC (aa451-459), were found to induce T cell proliferation in more than half of the patients tested. Of interest was that these three regions were also recognized by B cells. Information concerning human B and T cells epitopes can contribute greatly to the elucidation of the etiology of beef allergy.

Biochemical and Biophysical Research Communications


カルノシンおよびトリ胸肉抽出物(チキンエキス)のマウス遊泳疲労回復力に及ぼす効果

Journal of Japanese Society of Nutrition and Food Science


トリ胸肉抽出物(チキンエキス)のマウス遊泳持久力に対する効果

トリ胸肉抽出物(チキンエキス)のマウス遊泳持久力に対する効果


難消化性デキストリン配合ヨーグルト飲料の食後血糖値上昇に及ぼす影響

難消化性デキストリン配合ヨーグルト飲料の食後血糖値上昇に及ぼす影響


難消化性デキストリン含有ソーセージの便通改善効果

難消化性デキストリン含有ソーセージの便通改善効果


難消化性デキストリン含有ロースハムの便通改善効果

難消化性デキストリン含有ロースハムの便通改善効果


分離大豆タンパク質を添加したハンバーグ摂取による軽度高コレステロール血症を示す成人男子の血中コレステロール改善作用


Papain-Hydrolyzed Pork Meat Reduces Serum Cholesterol level and Premature Atherosclerosis in Dietary-Induced Hypercholesterolemic Rabbit

The effects of the low-molecular-weight fraction of papain-hydrolyzed pork meat (LMF) on the plasma cholesterol level and the generation of atherosclerosis were studied in rabbits fed a cholesterol-enriched diet. In LMF-fed rabbits, the plasma and liver cholesterol concentrations were both significantly lower (p<0.0 1) than in rabbits fed untreated pork meat (PM). Similarly, the cholesterol concentrations of the chylomicron and VLDL fractions were significantly lower in LMF-fed rabbits than in rabbits fed PM. Deposition of lipid in transverse sections of the aortic arch was significantly less in rabbits fed LMF than in those fed PM. Electron microscopic studies revealed preventive effects against premature atherosclerotic lesions in the aorta of rabbits fed LME These results indicate that LMF has a hypocholesterolemic action and preventive effects against premature atherosclerosis.

Journal of Nutritional Science and Vitaminology


アレルギー患者IgE抗体が結合する鶏肉タンパク質の同定

アレルギー患者IgE抗体が結合する鶏肉タンパク質の同定


IgE-antibody specificities of the patients allergic to meat products


高脂血症と食肉製品(大豆蛋白質)

高脂血症と食肉製品(大豆蛋白質)


Plasma cholesterol-suppressing effect of papain-hydrolyzed pork meat in rats fed hypercholesterolemic diet.

The effects of papain-hydrolyzed pork meat on plasma and liver cholesterol levels were studied in rats fed a cholesterol-enriched diet. In rats fed the low-molecular-weight fraction of papain-hydrolyzed pork meat, the plasma cholesterol concentration, more particularly the VLDL and LDL cholesterol concentrations, were significantly lower (p < 0.01) than in the rats fed untreated pork meat or soybean protein. Feeding with this fraction rather than with untreated pork meat also led to a significantly lower liver cholesterol concentration (p < 0.01) and increased fecal excretion of neutral and acidic steroids. The low-molecular-weight fraction contained peptides with molecular weights of 3,000 or less and had an amino acid composition similar to that of pork meat itself. This study suggests that peptides produced by papain-hydrolysis of pork meat have a hypocholesterolemic activity through their interference with the steroid absorption process.


Stimulation of Ethanol Metabolism Lnduced by Proline and Lysine Ingestion in Prolonged Ethanol-Administered Stroke-Prone Spontaneously Hypertensive Rats

Journal of Clinical Biochemistry and Nutrition


Effects of amino acids on alcohol intake in stroke-prone spontaneously hypertensive rats.

Examination was made of the effects of amino acids on alcohol intake in stroke-prone spontaneously hypertensive rats (SHRSP). The animals were divided into two groups according to dietary protein level, normal (15%) and low (5%). The two groups were further divided into two groups according to whether they selected the ethanol solution containing amino acids (non-amino acid group and amino acid group). The 5% ethanol solutions with and without various amino acids were prepared in a water-supplying tube to which the animals had free access for 40 days. The 5% ethanol solution intake in rats fed the normal protein diet was higher than that of the low protein group. Regardless of dietary protein level, 5% ethanol solution intake increased in the amino acid group. Intake of the 5% ethanol solution containing 100 mM L-proline, 100 mM L-lysine, and 100 mM L-threonine was large. For the amino acid group of rats fed normal protein and low protein, plasma glutamate oxalacetate transaminase (GOT) activity was significantly reduced. It is suggested that the alcohol intake may increase by adding amino acid to the alcohol solution.


Dietary Fatty Acid Ethyl Ester and Lecithin-Cholesterol Acyltransferase Activity in Rats


食肉タンパク質酵素加水分解物のラット血漿コレステロール上昇抑制効果

食肉タンパク質酵素加水分解物のラット血漿コレステロール上昇抑制効果


Nutritional Studies of the Roles of Dietary Protein levels and Umami in the Preference Response to Sodium Chloride for Experimental Animals

Previously we found that appetite or preference for sodium chloride depends not only on a genetic factor, but also on the nutritional status, i.e., dietary protein levels. To determine whether all kinds of protein had reducing effects on sodium chloride intake, the effect of dietary protein source on appetite or preference for sodium chloride was studied. It was found that the effect depended not only on protein levels, but also on the protein sources, i.e., amino acid patterns of diets. On the other hand, the turnover time of gustatory taste bud cells was measured by the pulse-labeled method using 3H-thymidine. It was found that the turnover time of gustatory cells in the taste buds of rats fed a low-protein diet was longer than in rats fed a high-protein diet. This suggested that some reduction of receptor functions may occur with low-protein diet. It was found that the dietary protein levels and umami also modified appetite or preference for alcohol solutions.


Structural Elcidation of Alterporriol B, a Novel Metabolic Pgment Produced by Alternaria porri (Ellis) Ciferri