Mie Prefecture Livestock Research Institute

Matsuzaka, Japan

Mie Prefecture Livestock Research Institute

Matsuzaka, Japan
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Han H.,Okayama University | Ogata Y.,Okayama University | Yamamoto Y.,Mie Prefecture Livestock Research Institute | Nagao S.,Okayama Prefecture Livestock Research Institute | Nishino N.,Okayama University
Journal of Dairy Science | Year: 2014

The survival of silage lactic acid bacteria (LAB) in the gut of dairy cows was evaluated by examining the LAB communities of silage and gut contents. Samples were collected at 2 different research institutes (Mie and Okayama) that offered total mixed ration (TMR) silage throughout the year. Silage and feces were sampled in August, October, and November at the Mie institute, whereas silage, rumen fluid, and feces were sampled in June and August at the Okayama institute. Denaturing gradient gel electrophoresis using Lactobacillus-specific primers was performed to detect LAB species in the samples. The selected bands were purified for species identification and the band patterns were used for principal component analysis. Lactic acid was the predominant fermentation product in all the TMR silages analyzed, and the lactic acid level tended to be constant regardless of the sampling time and region. A total of 14 LAB species were detected in the TMR silage samples, of which 5 (Lactobacillus acetotolerans, Lactobacillus pontis, Lactobacillus casei, Lactobacillus suebicus, and Lactobacillus plantarum) were detected in the dairy cow feces. Most of the denaturing gradient gel electrophoresis bands for the feces samples were also detected in the rumen fluid, suggesting that any elimination of silage LAB occurred in the rumen and not in the postruminal gut segments. The principal component analysis indicated that the LAB communities in the silage, rumen fluid, and feces were separately grouped; hence, the survival of silage LAB in the cow rumen and lower gut was deemed difficult. It was concluded that, although the gut LAB community is robust and not easily affected by the silage conditions, several LAB species can inhabit both silage and feces, which suggests the potential of using silage as a vehicle for conveying probiotics. © 2014 American Dairy Science Association.


PubMed | Okayama Prefecture Livestock Research Institute, Mie Prefecture Livestock Research Institute and Okayama University
Type: Journal Article | Journal: Journal of dairy science | Year: 2014

The survival of silage lactic acid bacteria (LAB) in the gut of dairy cows was evaluated by examining the LAB communities of silage and gut contents. Samples were collected at 2 different research institutes (Mie and Okayama) that offered total mixed ration (TMR) silage throughout the year. Silage and feces were sampled in August, October, and November at the Mie institute, whereas silage, rumen fluid, and feces were sampled in June and August at the Okayama institute. Denaturing gradient gel electrophoresis using Lactobacillus-specific primers was performed to detect LAB species in the samples. The selected bands were purified for species identification and the band patterns were used for principal component analysis. Lactic acid was the predominant fermentation product in all the TMR silages analyzed, and the lactic acid level tended to be constant regardless of the sampling time and region. A total of 14 LAB species were detected in the TMR silage samples, of which 5 (Lactobacillus acetotolerans, Lactobacillus pontis, Lactobacillus casei, Lactobacillus suebicus, and Lactobacillus plantarum) were detected in the dairy cow feces. Most of the denaturing gradient gel electrophoresis bands for the feces samples were also detected in the rumen fluid, suggesting that any elimination of silage LAB occurred in the rumen and not in the postruminal gut segments. The principal component analysis indicated that the LAB communities in the silage, rumen fluid, and feces were separately grouped; hence, the survival of silage LAB in the cow rumen and lower gut was deemed difficult. It was concluded that, although the gut LAB community is robust and not easily affected by the silage conditions, several LAB species can inhabit both silage and feces, which suggests the potential of using silage as a vehicle for conveying probiotics.


Fujita S.,Mie University | Okamoto R.,Mie University | Taniguchi M.,Mie University | Ban-Tokuda T.,Mie University | And 9 more authors.
Journal of Biochemistry | Year: 2013

Hibernation-specific protein (HP) is a plasma protein that regulates hibernation in chipmunks. The HP complex (HP20c) consists of three homologous proteins, HP20, HP25 and HP27, all produced by liver and belonging to the C1q family. To date, HP20c has not been identified in any mammalian species except chipmunk and ground squirrel hibernators. Here, we report a bovine HP20 gene isolated from liver tissue and aortic endothelial cells. Total homology between bovine and chipmunk variants was 63% at the amino acid level. Gene expression was highest in the liver. Western blot revealed HP20 protein in foetal, newborn, calf and adult serum, with highest concentrations in the adult. Similar proteins were detected in sera of other ruminants but not in humans, bears, mice or rats. Bovine HP20 protein was found mainly in ovaries, stomach, heart, kidneys, lungs, testes and prostate, but not in the skeletal muscle. Native HP20 was purified from bovine adult serum as a complex containing 25 and 27 kDa proteins. Mass spectrometry revealed that these proteins are orthologues of chipmunk HP25 and HP27, respectively. Interestingly, bovine HP20 was highly expressed in cattle serum after fasting. Native bovine HP20c may be a useful tool for investigating HP function. © 2013 The Authors 2013. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.


Kobayashi K.-I.,Toyohashi University of Technology | Matsui Y.,Mie Prefecture Livestock Research Institute | Maebuchi Y.,Toyohashi University of Technology | Nishino K.,Toyohashi University of Technology | And 2 more authors.
5th European Conference on Colour in Graphics, Imaging, and Vision and 12th International Symposium on Multispectral Colour Science 2010, CGIV 2010/MCS'10 | Year: 2010

The beef quality grade is greatly affected by visible fat content. Especially, in Japanese black (Wagyu) cattle, high fat content is typically valued highly In this paper, we describe the feasibility of beef evaluation by visualizing fat characteristics using near-infrared (NIR) multispectral imaging. An intact raw beef cut from Wagyu cattle was used as an evaluation target. The content of fat and fatty acid, such as the total saturated fatty acid (SFA) content, the total unsaturated fatty acid (UFA) content, myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), myristoleic acid (C14:1), palmitoleic acid (C16:1), oleic acid (C18:1), and linoleic acid (C18:2) were estimated and visualized. The total SFA content was calculated as the sum of myristic acid, palmitic acid, and stearic acid. Also, the total UFA content was calculated as the sum of myristoleic acid, palmitoleic acid, oleic acid, and linoleic acid. Reference values for the fat content and fatty acid composition were determined by conventional physical and chemical methods. The fatty acid composition was determined from the extracted lipids by Folch's method, by gas chromatography (GC) using its methyl ester. The fat content was determined by using the Gerhardt SOXTHERM. The NIR multispectral images of the sample were acquired by using the SPECIM Spectral Camera SWIR. It works in the wavelength range of 970-2500 nm with 6.3 nm of bandwidth at 320 pixels resolution in spatial domain. The absorbance spectra of each pixel calculated from pixel intensity of subject and reference white standard was used for constructing the prediction model. In total, 33 samples from various parts of the 2 head of Wagyu cattle were measured. Calibrations were performed by a partial least squares (PLS) regression using mean extracted spectra from each individual sample, limited wavelength range from 1000 to 2300 nm. The coefficients of determination (R2) were between 0.68 and 0.87. The ranks by evaluation index (EI) were "B (high accuracy)" and "C (slightly high)". The ratios of the standard error of prediction to the standard deviation (RPD) were between 1.74 and 2.74. These results indicate a sufficient feasibility of the prediction except for myristoleic acid content. The visualizations, which show the spatial distribution of fatty acid content, were performed by applying the model to predict the content of each pixel.


Hata T.,Mie University | Murakami K.,Nagoya University | Nakatani H.,Nagoya University | Yamamoto Y.,Mie Prefecture Livestock Research Institute | And 2 more authors.
Biochemical and Biophysical Research Communications | Year: 2010

By a series of centrifugation and ultracentrifugation, we could isolate microvesicles with approximately 100 nm in diameter from bovine milk. We also found that approximately 1700 and 1000 ng of total RNA, in which small RNAs were major components, was contained inside the microvesicles isolated from 6 ml of colostrum and mature milk, respectively, despite high RNase activity in the milk. Polyadenylated gene transcripts for major milk proteins and translation elongation factor-1α (EF-1α) were present in the microvesicles, and integrity of some transcripts was confirmed by real-time PCR targeting 5′- and 3′-ends of mRNA and by in vitro translation analysis. Moreover, a considerable amount of mammary gland and immune-related microRNAs were present in the milk-derived microvesicles. Acidification of milk to mimic gastrointestinal tract did not mostly affected RNA yield and quality. The milk related gene transcripts were detected in cultured cells when incubated with milk-derived microvesicles, suggesting cellular uptake of the microvesicle contents including RNA. Our findings suggest that bovine breast milk contains RNAs capable for being transferred to living cells and involved in the development of calf's gastrointestinal and immune systems. © 2010 Elsevier Inc. All rights reserved.


Kobayashi K.I.,Toyohashi University of Technology | Matsui Y.,Mie Prefecture Livestock Research Institute | Maebuchi Y.,Toyohashi University of Technology | Toyota T.,Toyohashi University of Technology | Nakauchi S.,Toyohashi University of Technology
Journal of Near Infrared Spectroscopy | Year: 2010

The meat quality grade of a beef carcass is greatly affected by its visible fat content. In premium beef from Japanese Black (Wagyu) cattle, a high fat content is greatly valued. However, the fatty acid composition, which is linked to the properties of the fat, is not considered in grading. In this paper, we describe the feasibility of an evaluation method based on food composition and its distribution. An intact raw beef cut from Wagyu cattle was used as an evaluation target. A total of 90 samples from various parts of three Wagyu cattle were measured by near infrared (NIR) hyperspectral imaging at wavelengths of 1000-2300 nm at a spatial resolution of 380 μm pixel-1 and were also analysed by conventional physical and chemical methods. The fat and fatty acid content were selected as the objective content, including the proportions of total saturated fatty acid (SFA), total unsaturated fatty acid (UFA) and the main fatty acids: myristic [C14:0, where Cx:y indicates the number of carbon atoms (x) and the number of double bonds (y)], palmitic (C16:0), stearic (C18:0), myristoleic (C14:1), palmitoleic (C16:1), oleic (C18:1) and linoleic (C18:2). The mean spectrum from an area extracted from the hyperspectral image to fit the area analysed by physical and chemical methods was used to develop partial least squares regression models for prediction of fat and fatty acid content. The prediction of total fat, SFA and UFA were satisfactory with r2, standard error of prediction (SEP) and ratio of prediction to deviation (RPD) values of 0.90, 0.87 and 0.89, 4.81%, 1.69% and 3.41% and 2.84, 2.43 and 2.84, respectively. For individual fatty acids, the r2 and RPD values ranged from 0.68 to 0.89 and 1.69 to 2.85, respectively. Prediction of fat content for each pixel of the hyperspectral image made using these prediction models yielded spatially distributed visualisations of the content. These results showed the feasibility of a beef evaluation method based on fat content evaluated by NIR hyperspectral imaging. © IM Publications LLP 2010.


Nishino N.,Okayama University | Ogata Y.,Okayama University | Han H.,Okayama University | Yamamoto Y.,Mie Prefecture Livestock Research Institute
Animal Science Journal | Year: 2015

As a forage source for total mixed ration (TMR) silage production, locally produced crop silage is now used in addition to imported hay. This type of TMR ensiling is regarded as a two-step fermentation process; hence, a survey was carried out to determine whether the bacteria in crop silage affect the subsequent TMR ensiling. Fermentation product contents and bacterial community were determined for TMR silage and its ingredient silages collected in August, October and November. August product contained corn, sorghum and Italian ryegrass silages, October product had wheat silage exclusively and November product did not include any crop silages. Acetic acid, lactic acid, 2,3-butanediol and ethanol were predominant fermentation products in corn, sorghum, Italian ryegrass and wheat silages, respectively. Robust lactic acid fermentation was seen in TMR silage, even if acetate-type and alcohol-type silages were mixed as ingredients. The finding that bacterial community of the TMR silage appeared unrelated to those of ingredient silage supported this. Silages of various fermentation types can therefore be formulated without interfering with lactate-type fermentation in TMR silage. © 2014 Japanese Society of Animal Science.


Sasaki K.,Mie Prefecture Livestock Research Institute | Tatsumi T.,Mie Prefecture Livestock Research Institute | Tsutsui M.,National Livestock Breeding Center Okazaki Station | Niinomi T.,National Livestock Breeding Center Okazaki Station | And 4 more authors.
Journal of Poultry Science | Year: 2010

A method of freezing semen using N-methylacetamide (MA) as a cryoprotective agent was applied to Yakido, a chicken breed designated as a "Natural Monument" under the "Law for the Protection of Cultural Property" (Law No. 214, May 30, 1950) in Japan. Semen was collected from Yakido roosters, and pooled semen was diluted 1: 1 using semen diluent containing no cryoprotective agent. After equilibration at 5°C for 30min, diluted semen was further diluted 1: 1 using semen diluent containing MA at a final concentration of 9%. Diluted semen was packaged into 0.5-mL plastic straws and frozen by placing the straws in liquid nitrogen vapor followed by plunging into liquid nitrogen at -196°C. After storage in liquid nitrogen for about one month, frozen semen was thawed in ice water. Intravaginal artificial insemination (AI) was performed immediately after thawing semen without removing the cryoprotective agent. In Fertility Trial 1, AI was performed once a week for four consecutive weeks. Weekly fertility rates using frozen semen were 83.8±3.8%, 77.1±8.8%, 88.6±5.1%, and 77.3±5.7% for weeks 1, 2, 3, and 4, respectively. Hatchability was over 90% throughout the experimental period. In Fertility Trial 2, the duration of fertility was examined after intravaginal insemination for 2 consecutive days. Daily fertility rates using frozen-thawed semen were 90.0%, 90.0%, 91.7%, 88.9%, 100.0%, 100.0%, 100.0%, 55.6%, 60.0%, 25.0%, 30.0%, 30.0%, and 9.1% for days 1 through 13 after AI, respectively. Overall hatchability was 89.5%. These results indicated that freezing chicken spermatozoa using MA as a cryoprotective agent is a practical method of preserving chicken semen collected from genetically valuable chicken breeds. © 2010 Japan Poultry Science Association.


Kobayashi K.-I.,Toyohashi University of Technology | Mori M.,Mie Prefecture Livestock Research Institute | Nishino K.,Toyohashi University of Technology | Toyota T.,Toyohashi University of Technology | Nakauchi S.,Toyohashi University of Technology
Journal of Near Infrared Spectroscopy | Year: 2012

Food quality is strongly affected by its components and their spatial distributions. Recently, spectroscopic methods have been widely applied as a non-destructive and rapid method to measure food quality. Although it is a versatile technique, the measurement system is extremely costly for practical use. In this paper, we propose a simple measurement system using a small set of band-pass filters. A food constituent was predicted using output from the band-pass filters as input for a multiple linear regression model, and the bands were designed to obtain high prediction accuracy characterised by the determination coefficient, using hyperspectral data by the optimisation approach. We designed three sets of filters to separately determine contents such as oleic acid, total unsaturated fatty acid and fat content in raw beef using NIR hyperspectral data, and then we implemented these designs as real optical filters. By mounting the filter in front of the lens of an NIR monochrome camera, we captured a set of filtered images. We then performed a pixel-by-pixel prediction of the content to enable the spatial distribution to be visualised. The determination coefficient (R2) and prediction error, which we characterised by the root mean square error of cross-validation (RMSECV), of this filtering method (R2 = 0.638-0.739, RMSECV = 3.13-5.15) were superior to those obtained with partial least squares (PLS) regression using hyperspectral measurements (R2 = 0.610-0.643, RMSECV = 3.70-6.12). Our method, therefore, facilitates the application of a hyperspectral technique for practical use. © IM Publications LLP 2012. All rights reserved.

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