Viral RNA extraction using an automatic nucleic acid extractor with magnetic particles and genetic characterization of bovine viral diarrhea virus in Tokachi Province,Japan, in 2016–2017

In this study, the viral genome extraction performance of automatic nucleic acid extractors and manual nucleic acid extraction kits was compared. We showed that compared with manual kits, the automatic extractors showed superior genome extraction performance using bovine viral diarrhea virus (BVDV) genome-positive cattle sera and bovine coronavirus/infectious bovine rhinotracheitis virus-spiked cattle nasal swabs. In addition, the subgenotyping of BVDV strains detected in Tokachi Province in Japan during 2016–2017 was performed. Results showed that most of these BVDV strains belonged to subgenotype 1b, while few strains belonged to subgenotypes 1a and 2a. This study showed the high applicability of automatic nucleic acid extractors in extracting multiple viral genomes and the dominant subgenotype of BVDV in Tokachi.     

The effects of long-term conservation tillage, crop residues and P fertilizer on soil conditions and responses of summer and winter crops on an Andosol in Japan

A field experiment was conducted from 1983 to 1992 in Tsukuba, Japan to investigate the effects of tillage on soil conditions and crop growth in a light-colored Andosol. Three tillage methods (NT: no-tillage, RT: no-tillage for summer cropping and moldboard plowing for winter cropping, and CT: conventional rotary tillage to a depth of 15 cm) were employed in combination with crop residue application (+R, −R) and fused magnesium phosphate (FMP) fertilization (+P, −P). Under the combination of NT and +R, diurnal variation of soil temperature at a depth of 5 cm was smaller during the summer cropping season and soil temperature in the daytime was lower during the winter cropping season than under CT. Soil inorganic N concentration at a depth of 0–30 cm was +R > −R and NT > RT > CT. The early growth of summer crops was accelerated under NT in comparison with CT, and yields were higher under NT and RT in comparison with CT. On the other hand, winter crop yields were significantly reduced under NT, while they were still higher under RT in comparison with CT. Yields were higher with +R and +P application, respectively, and these effects were more pronounced in winter cropping. The positive effect of FMP fertilization was greater in combination with NT, and that of residue treatment was greater in combination with RT and NT than with CT. In conclusion, the best tillage practice for Andosols on the Kanto Plain is RT, i.e. a combination of NT for summer cropping and CT for winter cropping. The application of NT for winter cropping is not recommended, although the application of phosphate and crop residues could reduce the risk of yield reduction, because of improved soil nutrient status and moderation of diurnal soil temperature.     

Study on the glass transition for several processed fish muscles and its protein fractions using differential scanning calorimetry

ABSTRACT:   The glass transition behavior of processed fish muscles (bonito, tuna, mackerel, sea bream, cod) and its muscle protein fractions (sarcoplasmic and myofibrillar proteins) were studied using differential scanning calorimetry. Each dried processed fish muscle and the extracted protein fractions showed clear glass transition phenomenon. The T _g values of muscles and myofibrillar proteins from red muscle fishes tended to be lower than those from white muscle fishes though there was no difference on T _g of sarcoplasmic proteins. The T _g value of whole muscle was considerably lower than that of extracted protein fractions because of the plasticizing effects of low molecular weight materials contained in the muscle.     

A retrospective study of inflammatory colorectal polyps in miniature dachshunds

Medical records of dogs with colorectal polyps were retrospectively reviewed, and clinical presentation of inflammatory colorectal polyps in miniature dachshunds was evaluated. Of 33 dogs found to have colorectal polyps, miniature dachshunds were markedly over-represented with 16 dogs (48%), of which 12 (75%) were found to have inflammatory polyps. Multiple polyps localized between the rectum and the descending colon was the most common finding in miniature dachshunds with inflammatory polyps. Twenty dogs (80%) out of 25 miniature dachshunds with inflammatory colorectal polyps responded to immunosuppressive therapy using prednisolone and cyclosporine. The results of this study indicate that miniature dachshunds are predisposed to develop inflammatory colorectal multiple polyps, for which immunosuppressive therapy may be a treatment option.     

Ligand-binding characteristics of feline insulin-binding immunoglobulin G

Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e⁻⁴ M for fraction B (low affinity IgGs) and 2e⁻⁵ M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin.     

A canine case of skull base meningioma treated with hydroxyurea

An 11-year-old female miniature schnauzer was tentatively diagnosed with the skull base meningioma, based on several examinations. Because surgical treatment was difficult, and outpatient radiation therapy was not available in the local area, chemotherapy with hydroxyurea combined with dexamethasone was selected. The patient's clinical symptoms improved after one week of treatment, and the tumor size was obviously reduced on MRI performed 37 days after treatment began. The patient received hydroxyurea for 7 months, with symptoms remaining stable, and the tumor re-increased to almost the same size at 7 months as that at the initial examination. At that time, hydroxyurea was discontinued. The patient died from pulmonary edema 14 months after treatment began. Pathologically, the tumor was diagnosed as a meningioma.     

Effect of propylene glycol and undegradable protein source on rumen fermentation, blood metabolism and milk production in lactating dairy cows

Eight lactating Holstein cows were divided into two groups ( n  = 4) and used in a double reversal trial with three periods of 14 days each, to evaluate diets containing propylene glycol (PG) and ruminally undegradable protein (RUP) blend on milk yield and composition, ruminal fermentation and blood metabolism. The control diet contained 20% chopped Sudangrass hay, 20% cubed alfalfa hay, 12% corn silage, and 48% of the respective concentrate mixtures (dry matter basis). The experimental diet (PG + RUP) partially replaced the concentrate mixture from the control diet with 1.4% PG and 2.1% RUP. Both diets contained about 16% crude protein and 71% total digestible nutrients. Dry matter intake was similar between the two diets. Daily production of milk, milk lactose and milk solids-not-fat increased by 9.0%, 11.3% and 9.3%, respectively ( P  < 0.1), for cows fed diets with PG + RUP; milk composition was unchanged. Although the concentration of ruminal total volatile fatty acids was unchanged, the proportion of propionic acid increased, and the proportion of acetic acid decreased with PG + RUP. The concentration of ruminal ammonia nitrogen and number of ciliate protozoa was not significantly affected by PG + RUP. The concentration of glucose in blood plasma increased, the concentration of urea nitrogen was unaffected, and the concentration of some essential free amino acids decreased with PG + RUP. It is suggested that these changes might be caused predominantly by PG, and the addition of PG may exert a favorable effect on milk production through increased metabolism.     

Regression tree analysis of the relationship between the concentrations of antimicrobial components and the microbiota of normal milk from dairy cows

The purpose of this study was to determine the concentrations of antimicrobial components (immunoglobulin A (IgA), lactoferrin (LF), lingual antimicrobial peptide (LAP), and S100A7) in normal milk and their relation to host factors (Age, somatic cell count (SCC), days in milk, richness, and alpha diversity of the milk microbiota) in dairy cows using multivariate regression tree analyses, and to clarify how the milk microbiota is related to the obtained results. Thirty normal milk samples were collected from a commercial dairy farm in June 2020. The thresholds that predicted the concentration of each antimicrobial component in milk were obtained by regression tree analysis, and the beta-diversity of the milk microbiota composition between groups divided according to each threshold was compared by an analysis of similarities test. The IgA and LF concentrations were mainly predicted by the SCC (177,500 and 70,000 cells/ml, respectively), and the LAP and S100A7 concentrations were predicted by Age (29.667 and 40.3 months, respectively). No relationship was observed between the concentration of IgA, LAP, or S100A7 and the milk microbiota composition between the groups divided by the threshold for prediction, but the milk microbiota composition was significantly different between the groups divided by the threshold for predicting the LF concentration. Our results indicated that the LF concentration in normal milk may be associated with the milk microbiota composition.