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.     

Cellulose digestion by common Japanese freshwater clam Corbicula japonica

ABSTRACT:   Cellulose digestion by Corbicula japonica was investigated according to the hypothesis that without any symbiotic aid, this organism can utilize cellulose as a carbon source. Enzymatic studies revealed the complete cellulase activity of this species, and molecular cloning resulted in the isolation of cDNA with an ORF encoding a 596-amino-acid protein that shares significant homology with abalone and termite cellulases with an amino acid identity of 52.2% and 50.5%, respectively. The isolated cellulase had a carbohydrate-binding module at the N-terminal region that was also reportedly present in abalone cellulase, and its mRNA were specifically expressed in the digestive gland. These findings strongly support the assumption that C. japonica has an endogenous cellulose, as well as abalones and termites. It is further believed that C. japonica plays an important roll in decomposing cellulose, and consequently contributes to the carbon-cycle in the aquatic environment, as termites do in terrestrial forests.     

Effect of dietary conjugated linoleic acid on lipid peroxidation and histological change in rat liver tissues

The effect of dietary conjugated linoleic acid (CLA) on hepatic lipid parameters in Sprague-Dawley rats was examined. When rats were fed a diet containing CLA at 0 (control), 1, or 2% of the weight of the amount of food given for 3 weeks, the liver weight exhibited a slight increase in the CLA-fed groups, although the difference was not significant. Lipid accumulation in the hepatocytes of CLA-fed rats was also demonstrated by electron microscopic observation. In addition, the liver thiobarbituric acid reactive substances levels were significantly higher in the 2 wt % CLA group than in the other two dietary groups, and the levels of phosphatidylcholine hydroperoxide were higher in CLA-fed groups when compared to that of the control group. On the other hand, the serum lipid peroxide levels were comparable among all three dietary groups. Levels of triglycerides in the white adipose tissue (WAT) and serum nonesterified fatty acid (NEFA) were reduced in a CLA-dose-dependent manner. CLA was shown to accumulate in the WAT much more than in the serum or liver. These results suggest that CLA accelerates the decomposition of storage lipids in WAT and the clearance of serum NEFA levels, resulting in lipid peroxidation and a morphological change in the liver.     

Production and analysis of reciprocal hybrids between <Emphasis Type="Italic">Asparagus officinalis</Emphasis> L. and <Emphasis Type="Italic">A. schoberioides</Emphasis> Kunth

The genus Asparagus (Asparagaceae) encompasses 100–300 species, including the important vegetable crop, A. officinalis (garden asparagus). Previous attempts to hybridize garden asparagus with A. densiflorus (Kunth) Jessop, with the aim of introducing disease resistance, were unsuccessful because of the failure of endosperm development. In the present study, reciprocal interspecific hybrids between garden asparagus and A. schoberioides Kunth were generated by hand pollination. The F1 hybrids were analyzed by using both morphological and molecular techniques. This is the first report describing the production of an interspecific hybrid between garden asparagus at the diploid level (2n = 2x = 20) and its diploid (2n = 2x = 20) wild relative, A. schoberioides. Morphological characteristics of candidate hybrids were a mixture of those found in the parents, and cytological and RFLP analyses confirmed that morphologically intermediate plants were indeed diploid hybrids of those two species. In other words, post-zygotic isolation is not complete between phylogenetically distinct these two species. Our results suggest that other Asparagus species may be capable of hybridizing with A. officinalis and that introducing wild characters through interspecific hybridization may offer advantages for breeding for novel traits. Takuro Ito and Toshinori Ochiai authors are contributed equally to this work     

Dependence of self-tolerance on TRAF6-directed development of thymic stroma

The microenvironments of the thymus are generated by thymic epithelial cells (TECs) and are essential for inducing immune self-tolerance or developing T cells. However, the molecular mechanisms that underlie the differentiation of TECs and thymic compartmentalization are not fully understood. Here we show that deficiency in the tumor necrosis factor receptor-associated factor (TRAF) 6 results in disorganized distribution of medullary TECs (mTECs) and the absence of mature mTECs. Engraftment of thymic stroma of TRAF6(-/-) embryos into athymic nude mice induced autoimmunity. Thus, TRAF6 directs the development of thymic stroma and represents a critical point of regulation for self-tolerance and autoimmunity.     

Regulation of myosin phosphatase by a specific interaction with cGMP- dependent protein kinase Ialpha

Contraction and relaxation of smooth muscle are regulated by myosin light-chain kinase and myosin phosphatase through phosphorylation and dephosphorylation of myosin light chains. Cyclic guanosine monophosphate (cGMP)-dependent protein kinase Ialpha (cGKIalpha) mediates physiologic relaxation of vascular smooth muscle in response to nitric oxide and cGMP. It is shown here that cGKIalpha is targeted to the smooth muscle cell contractile apparatus by a leucine zipper interaction with the myosin-binding subunit (MBS) of myosin phosphatase. Uncoupling of the cGKIalpha-MBS interaction prevents cGMP-dependent dephosphorylation of myosin light chain, demonstrating that this interaction is essential to the regulation of vascular smooth muscle cell tone.     

Occurrence and characterization of tomato spotted wilt orthotospovirus isolated from cucumber

European Journal of Plant Pathology - Diseases associated with tomato spotted wilt orthotospovirus (TSWV) are a serious threat to vegetable production worldwide. In 2019, leaf necrosis disease was...     

Reliable enzyme-linked immunosorbent assay for the determination of soybean proteins in processed foods

Among allergenic foods, soybean is known as a food causing adverse reactions in allergenic patients. To clarify the validity of labeling, the specific and sensitive detection method for the analysis of the soybean protein would be necessary. The p34 protein, originally characterized to be p34 as an oil-body associated protein in soybean, has been identified as one of the major allergenic proteins and named Gly m Bd 30K. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of the soybean protein in processed foods was developed using polyclonal antibodies raised against p34 as a soybean marker protein and the specific extraction buffer for extract. The developed sandwich ELISA method was highly specific for the soybean protein. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed ELISA were 0.47 ng/mL (equivalent to 0.19 microg/g in foods) and 0.94 ng/mL (equivalent to 0.38 microg/g in foods), respectively. The recovery ranged from 87.7 to 98.7%, whereas the intra- and interassay coefficients of variation were less than 4.2 and 7.5%, respectively. This study showed that the developed ELISA method is a specific, precise, and reliable tool for the quantitative analysis of the soybean protein in processed foods.     

Safety of reduced-dosage ketoprofen for long-term oral administration in healthy dogs

OBJECTIVE: To evaluate the safety of reduced-dosage ketoprofen (RDKET) for long-term oral administration in healthy dogs. ANIMALS: 14 healthy Beagles. PROCEDURES: Racemic ketoprofen (0.25 mg/kg, PO) and gelatin capsules, as a drug-free placebo, were each administered to 7 dogs for 30 days. Dogs were periodically monitored via physical examination, blood analyses, endoscopic examinations, fecal occult blood tests (tetramethylbenzidine and guaiac methods), renal function tests (effective renal plasma flow and glomerular filtration rate), urinalyses, urinary enzyme indices (N-acetyl-beta-D-glucosaminidase and gamma-glutamyl-transferase), and hemostatic function tests (buccal mucosa bleeding time, cuticle bleeding time, prothrombin time, activated partial thromboplastin time, and fibrinogen concentration). RESULTS: Pyloric antrum lesion grade was significantly higher in the RDKET group on day 28, compared with the pretreatment and control group grades. Fecal occult blood grade measured by use of the tetramethylbenzidine method was significantly higher in the RDKET group on day 30, compared with the pretreatment grade. No other significant differences were detected between treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: RDKET induced mild to moderate gastric mucosal injuries especially in the pyloric antrum in healthy Beagles, whereas no adverse effects were observed in renal function or hemostasis. Fecal occult blood tests may be useful as screening tests for adverse gastrointestinal effects induced by RDKET in dogs.     

Efficient digestion and structural characteristics of cell walls of coffee beans

Screening of effective food-processing cellulase for digestion of cell walls of coffee beans was carried out, and the cellulase from Trichoderma sp. was selected. The digestion of the cell walls of green and roasted coffee beans was carried out by sequential procedures of alkali boiling (0.1 M Na2CO3 buffer, pH 10, and 0.1 M NaOH), cellulase digestion, autoclaving with 0.1 M NaOH, and cellulase redigestion. The total digestion yields were >95 and >96%, respectively. The cell walls became thin, and the final residues of the cell walls were easily broken into small pieces. The neutral sugar analysis of the digestion or the extract and the residues and the microscopy observations with staining with toluidine blue O, Yariv reagent, and calcofluor for the residue in each step were investigated. Four structures, the galactomannan-cellulose (center part), the membrane of the arabinogalactan protein, the cellulose-rich galactomannan layer, and the arabinogalactan protein-rich layers (outer part), were found in the cell walls.