Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

Nutrient Digestibility of Cereal Grain Products and By‐products in Extruded Diets for Nile Tilapia

Apparent digestibility coefficients (ADCs) provide estimates of nutrient availability in feedstuffs and can be used as a tool for selecting ingredients to compound cost‐effective diets for fish. Extrusion process is one factor that could influence ADC values. Thus, a digestibility trial was undertaken to evaluate ADC of protein, energy, and dry matter, and apparent amino acid availability (AAAA) in five cereal grain products/by‐products (corn, wheat middlings, broken rice, rice bran, and sorghum) in compounded diets for Nile tilapia. Chromic oxide was used as an external inert marker. The digestibility values of energy and dry matter in ingredients, in decreasing order, were highest for broken rice (95.34 and 96.45%), sorghum (82.37 and 87.29%), corn (67.34 and 82.21%), rice bran (57.58 and 55.59%), and wheat middlings (48.94 and 45.88%). Protein digestibility value was higher for corn (72.86%) and the lowest value was observed for sorghum (56.77%), while broken rice, wheat middlings, and rice bran presented values ranging from 63.01 to 66.88%. AAAA values were higher for corn (70.05%) and the lowest methionine and cystine availability values were recorded for wheat middlings and broken rice, respectively. Broken rice appears as one of the most indicated ingredient for using in Nile tilapia diets because of its highest values of nutrient digestibility when compared to the other energetic sources. The use of this information should aid more precise and economic diet formulation for Nile tilapia.     

Purification and characterization of two anionic trypsins from the hepatopancreas of carp

SUMMARY: Two trypsins, designated as trypsin A and trypsin B, have been purified from the hepatopancreas of carp. The purification procedures consisted of ammonium sulfate fractionation, and chromatographies on DEAE-Sephacel, Ultrogel AcA54 and Q-Sepharose. Trypsin A was purified to homogeneity with the molecular mass of approximately 28 kDa, while trypsin B gave two close bands of 28.5 kDa and 28 kDa on sodium dodecylsulfate polyacrylamide gel electrophoresis both under reducing and non-reducing conditions. On native-PAGE, both trypsin A and trypsin B showed a single band. Trypsin A and trypsin B revealed optimum temperature of 40°C and 45°C, respectively, and shared the same optimum pH 9.0 using Boc-Phe-Ser-Arg-MCA as substrate. Both enzymes were effectively inhibited by trypsin inhibitors and their susceptibilities were similar. The NH₂-terminal amino acid sequences of trypsin A and trypsin B were determined to 37th and 40th amino acid residue, respectively. Their sequences were very homologous, but not identical to that of a trypsin-type serine proteinase from carp muscle and these of other trypsins. Immunoblotting test using the antibody raised against trypsin A cross-reacted with trypsin B positively.     

Trade-off evaluation using carbon dioxide equivalent and hazard index of a paddy soil with application of organic liquid fertilizer

<正>Dear Editor,Rice is an important staple food crop in Asia, but the anaerobic soil conditions in rice fields cause emission of the potent greenhouse gas, methane(CH₄)(Le Mer and Roger,2001), and promote the accumulation of arsenic(As) in the grain(Le Mer and Roger, 2001; Khanam et al., 2020).To facilitate sustainable agriculture, using wastewater as an organic fertilizer is essential for reducing wastewater treatment-associated energy consumption and the recycling of wastewate...     

Dietary apigenin suppresses IgE and inflammatory cytokines production in C57BL/6N mice

Flavonoids ubiquitously exist in plants, vegetables, fruits, and teas. We evaluated the effect of dietary apigenin, one of the well-known flavonoids, on the immune system in C57BL/6N mice. Mice were fed experimental diets containing apigenin for 2 weeks. After the experimental period, there was no significant difference in body and organ weights between the control and the apigenin group. The total immunoglobulin (Ig) E levels in mice fed apigenin were significantly suppressed, whereas levels of IgG, IgM, and IgA were not affected. We also examined the effect of the apigenin diet on cytokine expression in mice sera using a cytokine array. The production of regulated upon activation normal T cell expressed and secreted (RANTES) and soluble tumor necrosis factor receptor I (sTNFRI) in mice sera was down-regulated by the apigenin diet. These results suggest that a diet containing apigenin can reduce serum IgE and inflammatory cytokines such as RANTES and sTNFRI in mice.     

Rhizome-induction activity of chiral 2-α-methylbenzylamino-4-alkylamino-6-chloro-1, 3, 5-triazines in Cyperus serotinus Rottb.

A cytokinin-like effect of chiral 2-α-methylbenzylamino-4-alkylamino-6-chloro-1,3,5-triazines was found using a rhizome-inducing assay with Cyperus serotinus Rottb. tubers. C. serotinus tubers germinated in distilled water yielded plantlets with roots and leaves. Secondary rhizomes were normally not observed within the regular 14-day incubation time in water culture, whereas after increasing incubation periods a very short rhizome appeared (controls). 6-Benzylaminopurine (BA) significantly stimulated rhizome induction, while other plant hormones were inactive. The (R)-isomers of the 1, 3, 5-triazine compounds also stimulated induction of the rhizomes, whereas the (S)-isomers did not. The described rhizome induction system seems to be suitable as a cytokinin bioassay. The (R)-1, 3, 5-triazine compounds showing rhizome-inducing activity (RI activity) inhibited root formation and plant growth at high concentrations with symptoms which were very similar to those of BA. Therefore, the (R)-isomers appear to act as cytokinins in the rhizome induction assay.     

PCR detection of Porcine circovirus type 2 DNA in whole blood,serum, oropharyngeal swab,nasal swab,and feces from experimentally infected pigs and field cases

Porcine circovirus type 2 (PCV2) shedding patterns were investigated by polymerase chain reaction (PCR) for the detection of PCV2 DNA, and the diagnostic suitability of a sample for the PCR was examined by using different types of samples. In the experimental infection, sixteen pigs were inoculated intranasally with PCV2. The samples, including oropharyngeal and nasal swabs, feces, whole blood and serum became positive for PCV2 DNA by PCR immediately after the inoculation, and almost all samples remained positive during the observation period, post-inoculation-day 70. Field samples were collected from 313 pigs in five different age groups. The overall percentages of positive samples in the whole blood, nasal swabs, and feces detected by PCR were 30.4%, 19.2%, and 20.4%, respectively. The frequency of positive samples increased after the nursery stages and reached a peak in the 3 to 4-month-old pigs. These results indicate that PCV2 infection may occur after weaning, that PCV2 DNA may be present in whole blood for a long period after infection, and that whole blood and serum are the most suitable sample types for the PCR analysis of PCV2.     

Determination of nucleotide sequence of SRY gene in sika deer (Cervus nippon)

The aim of the present study was to determine the whole nucleotide sequence of the open reading frame of the sex‐determining region Y (SRY‐ORF) in wild sika deer. The SRY gene of wild sika deer was obtained by polymerase chain reaction (PCR) with DNA from blood samples. The whole nucleotide sequence of the SRY‐ORF in wild sika deer consisted of 687 bp and encoded 229 deduced amino acids. In comparison with the bovine SRY gene, the percentage of nucleotide sequence homology was 91.0% in the overall ORF, and those of the N‐terminal, high mobility group (HMG) box, and C‐terminal regions within ORF were 88.9%, 96.2% and 87.9%, respectively. The nucleotide sequences of sika deer SRY‐ORF characterized in the present study can be used for phylogenetic analysis or sexing in wild sika deer.