Survey of tick infestation and tick-borne ehrlichial infection of dogs in Ishigaki Island, Japan.

Twelve (54.5%) of 22 free-roaming dogs in Ishigaki Island had tick infestation identified as Rhipicephalus sanguineus. There were 121 ticks recovered and consisted of 28 females, 58 males, 22 nymphs and 3 larvae. Infection of dogs possibly with canine ehrlichial pathogens was examined by both indirect immunofluorescence assay and polymerase chain reaction (PCR). Two dogs of the 13 examined were sero-positive for the human granulocytic ehrlichia agent, and one of two dogs was PCR positive for Ehrlichia platys. This dog had platelet numbers slightly lower than normal value, however, no morulae were found within platelet on peripheral blood smear stained with Giemsa.     

Immunohistochemical study on the delayed progression of epithelial apoptosis in follicle-associated epithelium of rat Peyer's patch

It is well known that some caspases in apoptosis is involved in determinant of terminal differentiation and maturation of various cells. Our previous study ultrastructurally clarified the differentiation into M cells from immature microvillous epithelial cells and the redifferentiation from M cells to microvillous epithelial cells in the follicle-associated epithelium (FAE) of rat Peyer's patch. In this study, the difference of epithelial apoptosis between the FAE of Peyer's patch and intestinal villi was immunohistochemically investigated in rat jejunoileum. As a result, cleaved caspase-3 was limited to several epithelial cells at the tip of FAE, whereas almost all of the epithelial cells were cleaved caspase-3 positive in intestinal villi. Cleaved caspase-9 was detected only in a few exfoliating or exfoliated epithelial cells of both FAE and intestinal villi. Nuclear DNA-fragmentation was detected only in several epithelial cells of the tip of FAE, while it was expressed from the middle regions in the intestinal villi. The DNase I expression of the epithelial cytoplasm was much weaker in FAE than in intestinal villi. Bcl-x expression was restricted in the apical cytoplasms of epithelial cells in the FAE, whereas it was restricted in whole cytoplasms in villous epithelial cells. These findings suggest that the progression of the apoptotic process in the epithelial cells of FAE is later than in the intestinal villi, so that the possibility of epithelial differentiation might be remained in the FAE, unlike in the intestinal villi.     

Isolation and chemiluminescent properties of ferret (Mustela putorius furo) polymorphonuclear cells

Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formyl-Met-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets.     

Phylogenetic characterization of rabies virus isolates from Carnivora in Brazil

The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.     

Binding profiles and cytokine-inducing effects of fish rhamnose-binding lectins on Burkitt’s lymphoma Raji cells

Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt’s lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing β-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-α but not of IFN-γ, IL-1β, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-α-mediated signaling pathway.     

Fruit antioxidant activity, ascorbic acid, total phenol, quercetin, and carotene of Irwin mango fruits stored at low temperature after high electric field pretreatment

Greenhouse-grown tree ripe (TR) and mature green (MG) mangoes (cv. Irwin) were exposed to high electric field treatment before 20 and 30 days of storage at 5 degrees C. MG fruits were allowed to ripen at room temperature after low-temperature storage. Fruit physical quality attributes, ascorbic acid, carotene, quercetin, total phenols, and antioxidant capacity were estimated before and after the storage period. Antioxidant capacity of fruit juice was estimated using the ferric reducing antioxidant power (FRAP) assay. Fruit firmness decreased significantly during storage. Titratable acidity decreased 20 days after storage. Total soluble solids did not change during storage. Antioxidant capacity of fruits remained unchanged up to 20 days of storage period and decreased thereafter. Total phenol and carotenes increased during storage. Antioxidant capacity of fruits was significantly correlated only to ascorbic acids. Peel color and carotenes were higher in TR fruits, whereas titratable acidity and firmness were higher in MG fruits. There was no significant difference in other parameters between the stages of picking. Electric field pretreatment affected the respiration and antioxidant capacity of TR fruits and did not have any significant affect on other parameters. TR mangoes of cv. Irwin are more suitable for low-temperature storage and can be successfully stored for up to 20 days at 5 degrees C without any significant losses in functional properties and quality attributes.     

Effects of salivary gland extract from Rhipicephalus sanguineus on IgG subclass production and cytokine mRNA expression in mononuclear cells of canine peripheral blood

The effects of salivary gland extract (SGE) from R. sanguineus were examined on the production of IgG1 and IgG2 and the mRNA expression of IFN-gamma, IL-2, IL-4, IL-5 and IL-10 in the mononuclear cells from canine peripheral blood, treated with concanavalin A (ConA) in vitro. SGE suppressed the ConA-induced production of IgG2. It also inhibited the expression of IFN-gamma, IL-2 and IL-5 mRNA in a dose-dependent manner. No dose-dependent suppression was observed of IL-10 mRNA expression although a significant effect was observed at a SGE protein concentration of 25 microg/ml. SGE had no effect on the mRNA expression of IL-4. These results suggest that the suppression of IgG2 production by SGE from R. sanguineus was caused by the suppression of IFN-gamma production.     

Bovine milk enhances the oxidative burst activity of polymorphonuclear leukocytes in low concentrations

Bovine milk contains various immunoreactive components, and the activation of polymorphonuclear leukocytes (PMNLs) function in breast-fed infants has been reported. In this study, the effect of milk on the oxidative burst of bovine PMNLs was investigated in vitro. When PMNLs were incubated with 0.1% colostrum or normal milk, the oxidative burst induced by serum-opsonized Staphylococcus aureus was enhanced, and the enhancement declined dose-dependently. The enhancement of the oxidative burst by milk was not due to opsonins but the priming activities. Also, the phorbol 12-myristate 13-acetate (PMA)-induced oxidative burst increased after incubation with 0.1% colostrum, but the colostral enhancement of the oxidative burst was unaffected by the incubation time. These results suggest that bovine milk contains oxidative burst promoting factor(s).     

Root parameters of forest trees as sensitive indicators of acidifying pollutants: a review of research of Japanese forest trees

The root parameters of forest trees can be indicators of a changing environment. We summarize the results of root studies with regard to the effects of acidifying pollutants, especially soil acidification and aluminum toxicity, on various root parameters of Japanese forest trees under experimentally controlled conditions. All root parameters such as biomass, morphology, nutritional status, and physiology can be regarded as indicators, because, under laboratory conditions, root responses occur prior to the responses in the aboveground parts. However, considering the conditions of forest sites, the nutritional status and physiological changes are better indicators of soil acidification and Al stress than the biomass and morphological response. The currently available data suggest that the most important indicator is the Ca/Al molar ratio in roots of Japanese tree species. In order to predict and detect the initial effects of soil acidification, we postulate that the specific root response to the Ca/Al molar ratio of tree roots should be considered as a parameter for use in long-term forest monitoring sites.     

Estimating the production and mortality of fine roots in a Japanese cedar (Cryptomeria japonica D. Don) plantation using a minirhizotron technique

Fine roots are a key component of forested ecosystems, but available information is still limited. This study examined the production and mortality of fine roots less than 1 mm in diameter in a Japanese cedar (Cryptomeria japonica D. Don) plantation located on the Kanto Plain in central Japan. We used a minirhizotron technique in combination with soil coring, and collected data for 1 year (May 2002–May 2003). Fine root production and mortality were determined from changes in the lengths of individual fine roots on minirhizotron tubes. Both fine root production and mortality rates were greater in the upper soil than in lower soil levels. Both rates were seasonal, with higher values in summer than in winter; this trend was more pronounced in upper soil levels. These results suggest that environmental conditions, such as temperature or soil properties, affect the production and mortality rates of fine roots. Fine root production and mortality occurred simultaneously, and their rates were similar, which may have led to unclear seasonal changes in fine root standing crop estimates. Soil coring indicated that the fine root biomass of this stand was about 120 g m⁻², of which 40% was from Japanese cedar. The estimated rates of dry matter production and mortality of total fine roots, including understory plants, were both approximately 300 g m⁻² year⁻¹.