Metabolomic Analyses of Blood Plasma after Oral Administration of D-Glucosamine Hydrochloride to Dogs

D-Glucosamine hydrochloride (GlcN∙HCl) is an endogenous amino monosaccharide synthesized from glucose that is useful in the treatment of joint diseases in both humans and animals. The aim of this study was to examine amino acid metabolism in dogs after oral administration of GlcN∙HCl. Accelerated fumarate respiration and elevated plasma levels of lactic acid and alanine were observed after administration. These results suggest that oral administration of GlcN∙HCl induces anaerobic respiration and starvation in cells, and we hypothesize that these conditions promote cartilage regeneration. Further studies are required to evaluate the expression of transforming growth factor-beta (TGF-β).     

Effects of Oral Administration of Fucoidan Extracted from Cladosiphon okamuranus on Tumor Growth and Survival Time in a Tumor-Bearing Mouse Model

We evaluated the anti-tumor activities of the oral administration of fucoidan extracted from Cladosiphon okamuranus using a tumor (colon 26)-bearing mouse model. The materials used included low-molecular-weight fucoidan (LMWF: 6.5–40 kDa), intermediate-molecular-weight fucoidan (IMWF: 110–138 kDa) and high-molecular-weight fucoidan (HMWF: 300–330 kDa). The IMWF group showed significantly suppressed tumor growth. The LMWF and HMWF groups showed significantly increased survival times compared with that observed in the control group (mice fed a fucoidan-free diet). The median survival times in the control, LMWF, IMWF and HMWF groups were 23, 46, 40 and 43 days, respectively. It was also found that oral administration of fucoidan increased the population of natural killer cells in the spleen. Furthermore, from the results of the experiment using Myd-88 knockout mice, it was found that these effects are related to gut immunity. These results suggest that fucoidan is a candidate anti-tumor functional food.     

Metabolomic Analysis of Blood Plasma after Oral Administration of N-acetyl-d-Glucosamine in Dogs

N-acetyl-d-glucosamine (GlcNAc) is a monosaccharide that polymerizes linearly through (1,4)-β-linkages. GlcNAc is the monomeric unit of the polymer chitin. GlcNAc is a basic component of hyaluronic acid and keratin sulfate found on the cell surface. The aim of this study was to examine amino acid metabolism after oral GlcNAc administration in dogs. Results showed that plasma levels of ectoine were significantly higher after oral administration of GlcNAc than prior to administration (p < 0.001). To our knowledge, there have been no reports of increased ectoine concentrations in the plasma. The mechanism by which GlcNAc administration leads to increased ectoine plasma concentration remains unclear; future studies are required to clarify this mechanism.     

Protective Effect of Chitin Urocanate Nanofibers against Ultraviolet Radiation

Urocanic acid is a major ultraviolet (UV)-absorbing chromophore. Chitins are highly crystalline structures that are found predominantly in crustacean shells. Alpha-chitin consists of microfibers that contain nanofibrils embedded in a protein matrix. Acid hydrolysis is a common method used to prepare chitin nanofibrils (NFs). We typically obtain NFs by hydrolyzing chitin with acetic acid. However, in the present study, we used urocanic acid to prepare urocanic acid chitin NFs (UNFs) and examined its protective effect against UVB radiation. Hos: HR-1 mice coated with UNFs were UVB irradiated (302 nm, 150 mJ/cm²), and these mice showed markedly lower UVB radiation-induced cutaneous erythema than the control. Additionally, sunburn cells were rarely detected in the epidermis of UNFs-coated mice after UVB irradiation. Although the difference was not as significant as UNFs, the number of sunburn cells in mice treated with acetic acid chitin nanofibrils (ANFs) tended to be lower than in control mice. These results demonstrate that ANFs have a protective effect against UVB and suggest that the anti-inflammatory and antioxidant effects of NFs influence the protective effect of ANFs against UVB radiation. The combination of NFs with other substances that possess UV-protective effects, such as urocanic acid, may provide an enhanced protective effect against UVB radiation.     

Milky hemolymph syndrome associated with an intranuclear bacilliform virus in snow crab <Emphasis Type="Italic">Chionoecetes opilio</Emphasis> from the Sea of Japan

Milky hemolymph syndrome (MHS) is a previously undescribed disease in snow crab Chionoecetes opilio. Outbreaks of this disease occurred in reared and wild populations from the Sea of Japan when the water temperature was 2–3°C. The common symptom of the disease was a distinct milky or opaque coloration of the hemolymph. Some severely affected crabs also showed yellow or ivory discoloration on the ventral shell and uncalcification of the arthrodial membranes of walking legs. Histopathological changes consisted of widespread cellular degeneration characterized by enlarged nuclei with marginal hyperchromatosis and basophilic intranuclear inclusions in the interstitial connective tissues of various organs. Electron microscopy analyses of degenerated cells revealed a nonoccluded, enveloped bacilliform virus within the nuclei. The virus morphologically resembles white spot syndrome virus (WSSV) or unassigned intranuclear bacilliform viruses (IBVs) reported from other crustaceans. However, we could not detect WSSV from diseased tissues by PCR using WSSV-specific primers, and the tissues targeted by the virus differed from those targeted by IBVs. This is the first report of a virus from the genus Chionoecetes. Thus, we have tentatively designated the virus presumably associated with MHS as Chionoecetes opilio bacilliform virus (CoBV) until its relationship with other crustacean viruses can be clarified.     

A porcine lymphoma-derived cell line co-expressing IgM,IgG and IgA

A cell line (PL38PB) was established from blood samples of a 6-month-old pig that was diagnosed with lymphoma with CD5 expression. Histopathological examination revealed neoplastic lesions in the spleen, liver and lymph nodes. Tumor cells were immunohistochemically positive for CD20 and immunoglobulin heavy chains (μ, γ and α). Membranous CD5 and cytoplasmic Immunoglobulin M (IgM), ​Immunoglobulin G (IgG) and ​Immunoglobulin A (IgA) were detected in PL38PB cells by flow cytometry. In addition, the cytoplasm of PL38PB cells were positive for IgM, IgG and IgA by immunofluorescent. However, no Ig secretion was detected in culture supernatant by Ouchterlony gel diffusion method. Results suggest that PL38PB cells express three Ig isotypes that are produced but not secreted.     

Tissue retention and adverse reaction after intravenous injection of hematoporphyrin derivatives in dogs

We evaluated changes in hematology and chemical profile, and the tissue retention of hematoporphyrin derivative (HpD) following the intravenous injection in dogs. HpD at concentrations of 1, 5, 10, and 15 mg/kg was intravenously injected to 16 dogs (n=4 each) and complete blood count (CBC) and blood chemistry were performed on days 1, 3, 5, and 7 after the injection. To examine tissue retention, HpD (5 mg/kg) was administered to 15 dogs and 3 dogs were euthanized on days 1, 2, 7, 14, and 28 after the injection, respectively, to collect the skin, muscle, small intestine, spleen, kidney and liver as tissue samples. There were no significant changes in CBC and blood chemical profile except for an increase in LDH concentrations in dogs given 10 and 15 mg/kg of HpD at day 3. The levels of HpD retention in the tissues were ranked in the following order: liver > kidney > spleen > intestine > muscle > skin.     

Effects of chitin and chitosan on collagen synthesis in wound healing

Collagen synthesis was evaluated by measuring prolyl hydroxylase (PHL) activity induced within rat granulation tissue by a polyester non-woven fabric (NWF, 1 x 1 cm, 0.6 mm in thickness) impregnated with a chitin or chitosan suspension ranging in concentration from 0.1 to 10 mg/ml. In addition, PHL activity induced in rat granulation tissue by a NWF impregnated with a phosphate buffer solution was examined as a control. The PHL activity in each group remained low until 4 days post-implantation (Day 4). However, in the 10 mg chitin group, the PHL activity increased rapidly without scatter of the data at Day 7 and remained at a plateau until Day 14. In other groups, PHL activity increased linearly until Day 14. The data varied widely at Day 7. Compared to chitosan, chitin at the higher concentration was found to induce stable collagen synthesis in the early wound healing process.     

Aspergillus fumigatus infection in an ostrich (Struthio camelus)

An 11-month-old female ostrich (Struthio camelus) had become gradually emaciated over a 2-week period and subsequently died. Necropsy revealed white to green mold growth on the walls of caseous thickened air sac membranes and multiple white necrotic foci in the lungs and liver. Histologically, the multiple exudative, necrotic and granulomatous lesions were compatible with mycotic infection in the air sacs and lungs, and hyphae positively reacted with a monoclonal antibody (Mab-WF-AF-1) to Aspergillus fumigatus wall fractions. Multifocal hepatic necrosis was also found, and several spores were observed in the blood vessels. Fungal culture of these lesions yielded pure growth of A. fumigatus. This is an established case of fatal A. fumigatus infection in an ostrich reared in Japan.     

Evaluation of a sensor-based system of motion analysis for detection and quantification of forelimb and hind limb lameness in horses

OBJECTIVE: To compare a sensor-based accelerometer-gyroscopic (A-G) system with a video-based motion analysis system (VMAS) technique for detection and quantification of lameness in horses. ANIMALS: 8 adult horses. PROCEDURE: 2 horses were evaluated once, 2 had navicular disease and were evaluated before and after nerve blocks, and 4 had 2 levels of shoe-induced lameness, alternatively, in each of 4 limbs. Horses were instrumented with an accelerometer transducer on the head and pelvis, a gyroscopic transducer on the right forelimb and hind feet, and a receiver-transmitter. Signals from the A-G system were collected simultaneously with those from the VMAS for collection of head, pelvis, and right feet positions with horses trotting on a treadmill. Lameness was detected with an algorithm that quantified lameness as asymmetry of head and pelvic movements. Comparisons between the A-G and VMAS systems were made by use of correlation and agreement (kappa value) analyses. RESULTS: Correlation between the A-G and VMAS systems for quantification of lameness was linear and high (r2 = 0.9544 and 0.8235 for forelimb and hind limb, respectively). Quantification of hind limb lameness with the A-G system was higher than measured via VMAS. Agreement between the 2 methods for detection of lameness was excellent (kappa = 0.76) for the forelimb and good (kappa = 0.56) for the hind limb. CONCLUSIONS AND CLINICAL RELEVANCE: The A-G system detected and quantified forelimb and hind limb lameness in horses trotting on the treadmill. Because the data are collected wirelessly, this system might be used to objectively evaluate lameness in the field.