Effects of intravenous infusion of hypotonic lactated Ringer's solution on calves with diarrhea

The effects of intravenous infusion of hypotonic lactated Ringer's (LR: n=14) on plasma volume and venous blood gases were compared to those of hypotonic Ringer's solutions (RS: n=7) in diarrheic Japanese Black breed calves with metabolic acidosis. Venous blood samples were collected immediately before and after, and at 24 hr after the fluid infusion therapy. The LR and RS infusions increased relative plasma volume to 147.1 +/- 25.5% and 134.2 +/- 18.6%, respectively, just after the fluid therapy. The LR infusion induced an increase in the BE value (+5.1 +/- 4.8 mM) at 24 hr compared to that of RS. LR infusion should be explored as a treatment for dehydration and moderate metabolic acidemia caused by naturally occurring diarrhea in calves.     

Changes in blastogenic responses of lymphocytes and delayed type hypersensitivity responses after vaccination in dogs.

To clarify the immunologic effects of vaccination in dogs, we monitored total leukocyte and lymphocyte counts, humoral antibody responses, blastogenic responses of lymphocyte, and delayed type hypersensitivity (DTH) responses after vaccination. Mixed vaccines were administered on day 0 except for canine parvovirus (CPV) vaccine which was readministered on day 21. The puppy and adult dogs had a significant decrease in leukocyte and lymphocyte counts on day 7. The puppies showed a significant increase in the blastogenesis of lymphocytes after each vaccination, whereas the adult dogs had no significant changes. However, the adult dogs were divided into two groups, high responders and low responders in blastogenesis of lymphocytes. The dogs with higher or lower response in SI values on day 0 tended to show decrease or increase after the first vaccination, respectively. Since almost all dogs developed high titers of humoral antibody, it is considered that vaccination acts in an immunomodulative fashion. DTH responses to phytohemagglutinin (PHA) and CPV vaccine monitored at 0, 3, and 8 weeks after the first vaccination produced strong reactions, in particular those to CPV vaccine rose significantly after vaccination and maintained the higher responses for at least 2 months. These results suggest that DTH responses to PHA and CPV vaccine are helpful to monitoring non-specific and specific immune functions in vivo, therefore, DTH could be used as simple and rapid immunologic tests in canine practice.     

Development of a method for detecting Coxiella burnetii in cheese samples

Coxiella burnetii is the causative agent of Q fever, and the main route of infection in humans is inhalation of contaminated aerosols. Although oral transmission by contaminated raw milk or dairy products is also a possible route of human infection, there have been few studies investigating the presence of C. burnetii in dairy products. We developed a new method of extracting DNA from cheese and detecting C. burnetii DNA in cheese samples with a nested PCR assay. The limit of detection was 6.0 × 10(2) C. burnetii particles per gram. We subsequently used this method to examine the presence of C. burnetii in cheese at commercial markets in Tokyo from June 2005 to December 2008. Twenty-eight of 147 cheese samples were found to be positive for C. burnetii DNA. However, when we assessed the viability of C. burnetii by inoculating mice with DNA-positive samples, all of the samples were found to be negative. Thus, the viability of C. burnetii appears to have been lost in these cheese samples.     

Quality evaluation and prediction of Citrullus lanatus by 1H NMR-based metabolomics and multivariate analysis

(1)H NMR spectrometry in combination with multivariate analysis was considered to provide greater information on quality assessment over an ordinary sensory testing method due to its high reliability and high accuracy. The sensory quality evaluation of watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) was carried out by means of (1)H NMR-based metabolomics. Multivariate analyses by partial least-squares projections to latent structures-discrimination analysis (PLS-DA) and PLS-regression offered extensive information for quality differentiation and quality evaluation, respectively. The impact of watermelon and rootstock cultivars on the sensory qualities of watermelon was determined on the basis of (1)H NMR metabolic fingerprinting and profiling. The significant metabolites contributing to the discrimination were also identified. A multivariate calibration model was successfully constructed by PLS-regression with extremely high reliability and accuracy. Thus, (1)H NMR-based metabolomics with multivariate analysis was considered to be one of the most suitable complementary techniques that could be applied to assess and predict the sensory quality of watermelons and other horticultural plants.     

Immunohistochemical detection of phosphatidylserine and thrombospondin on denucleating erythroblasts in rat bone marrow

The hypothesis that apoptotic factors play some roles in the denucleation of erythroblasts has been confirmed by the immunohistological detection of both phosphatidylserine and thrombospondin as phagocytosis-inducing factors in general apoptotic events. Both phosphatidylserine and thrombospondin were detected on the surface of cell membrane of mature erythroblasts, while thrombospondin was also detected in more immature erythroblasts. The intensities of their immune reactions increased as the erythroids matured. During denucleation, the positivities of both phosphatidylserine and thrombospondin were restricted on the surface of the cell membrane surrounding the protruding nuclei. Thus, the apoptotic process involves denucleation of erythroblasts and phosphatidylserine, and thrombospondin acts as phagocytosis-inducing factors in the denucleation event.     

Antimicrobial susceptibilities of Listeria monocytogenes isolated in Japan

The antimicrobial susceptibility of 201 Listeria monocytogenes isolates from foods, environments, animals and human patients in Japan was determined. All isolates were susceptible to ampicillin, the first choice of drug for listeriosis treatment, chloramphenicol, dihydrostreptomycin, erythromycin, enrofloxacin, gentamicin, kanamycin, lincomycin, nosiheptide, salinomycin, vancomycin, and virginiamycin. A human strain was resistant to oxytetracycline. The Minimum Inhibitory Concentration (MIC) for 50% of the strains and the MIC for 90% of the strains were comparable in all the isolates. This is the first investigation to compare antibiotic resistances between isolates from foods and isolates from human patients in Japan. The result showed that most of the isolates were susceptible to antibiotics used in this study.     

Immunosuppression in dogs by pretransfusions with ultraviolet (UV)-irradiated whole blood.

Ultraviolet (UV)-irradiated whole dog blood prevented the initiation of proliferative responses in allogeneic mixed lymphocyte reactions (MLR). When dogs were given 4 weekly intravenous transfusions of UV-irradiated allogeneic donor whole blood, peripheral blood lymphocytes (PBL) of the recipients responded less significantly to donor PBL in MLR for over one week after the final transfusion. Red blood cell (RBC)-crossmatching of these dogs was negative. Dogs treated with UV-irradiated blood did not produce anti-donor PBL antibody, or IgG, IgM and C3 determined by the indirect Coombs test. These dogs also had negligible delayed type hypersensitivity (DTH) responses to donor PBL at the end of the treatment period, recognized as slightly suppression of skin graft rejection. In contrast, six dogs receiving injections of untreated allogeneic whole blood evidenced high proliferation in MLR, agglutination in RBC-crossmatching, acute rejection of skin grafts and three of them produced C3 in the Coombs test.     

Prediction of Japanese green tea ranking by gas chromatography/mass spectrometry-based hydrophilic metabolite fingerprinting

An innovative technique for green tea's quality determination was developed by means of metabolomics. Gas-chromatography coupled with time-of-flight mass spectrometry and multivariate data analysis was employed to evaluate the quality of green tea. Alteration of green tea varieties and manufacturing processes effects a variation in green tea metabolites, which leads to a classification of the green tea's grade. Therefore, metabolic fingerprinting of green tea samples of different qualities was studied. A set of ranked green tea samples from a Japanese commercial tea contest was analyzed with the aim of creating a reliable quality-prediction model. Several multivariate algorithms were performed. Among those, the partial least-squares projections to latent structures (PLS) analysis with the spectral filtering technique, orthogonal signal correction (OCS), was found to be the most practical approach. In addition, metabolites that play an important role in green tea's grade classification were identified.     

Incorporation of 15N-labeled ammonia into glutamine amide groups by protein-glutaminase and analysis of the reactivity for α-lactalbumin

Protein-glutaminase (PG) is an enzyme that catalyzes the deamidation of protein-bound glutamine residues. We found that an enzyme labeling technique (ELT), which is a stable isotope labeling method based on transglutaminase (TGase) reaction, is applicable for PG. PG catalyzed incorporation of (15)N-labeled ammonium ions into reactive glutamine amide groups in α-lactalbumin similarly to TGase and deamidated the most reactive glutamine amide group once labeled with (15)N. Furthermore, we investigated the effect of ammonium ions on the PG activity by peptide mapping, and more reactive glutamine residues were detected than were detected by the ELT in the presence of ammonium ions. This is probably because ammonium ions are competitive inhibitors, causing decreased reactivity for glutamine residues. We propose the reaction scheme of PG in the presence of the (15)N-labeled ammonium ions and show that the ELT method with PG is useful for evaluating the activity of PG.