Phenotypic and functional analysis of bovine peripheral blood dendritic cells before parturition by a novel purification method

Dendritic cells (DCs) are specialized antigen presenting cells specializing in antigen uptake and processing, and play an important role in the innate and adaptive immune response. A subset of bovine peripheral blood DCs was identified as CD172a⁺/CD11c⁺/MHC (major histocompatibility complex) class II⁺ cells. Although DCs are identified at 0.1%–0.7% of peripheral blood mononuclear cells (PBMC), the phenotype and function of DCs remain poorly understood with regard to maintaining tolerance during the pregnancy. All cattle used in this study were 1 month before parturition. We have established a novel method for the purification of DCs from PBMC using magnetic‐activated cell sorting, and purified the CD172a⁺/CD11c⁺ DCs, with high expression of MHC class II and CD40, at 84.8% purity. There were individual differences in the expressions of CD205 and co‐stimulatory molecules CD80 and CD86 on DCs. There were positive correlations between expression of cytokine and co‐stimulatory molecules in DCs, and the DCs maintained their immune tolerance, evidenced by their low expressions of the co‐stimulatory molecules and cytokine production. These results suggest that before parturition a half of DCs may be immature and tend to maintain tolerance based on the low cytokine production, and the other DCs with high co‐stimulatory molecules may already have the ability of modulating the T‐cell linage.     

Exfoliation rate of mammary epithelial cells in milk on bovine mastitis caused by Staphylococcus aureus is associated with bacterial load

The exfoliation rate of mammary epithelial cells (MECs) in milk is affected by physiological, breeding and environmental factors. Little is known about the relationship between the MEC exfoliation into milk and mammary‐infected Staphylococcus aureus (S. aureus) load on bovine mastitis caused by S. aureus. The aim of this study was to investigate the relationship between S. aureus load and the proportion of MEC exfoliation in milk using five substantial bovine mastitis models. In 64 randomly extracted milk samples from udders at 3–21 days after S. aureus infusion, there were various samples with different numbers of S. aureus counts and somatic cell counts. No significant correlations were found between the S. aureus counts and somatic cell count (r = 0.338). In contrast, a significant correlation was noted between S. aureus counts and the proportion of cytokeratin‐positive cells in the milk from the infused udders (r = 0.734, P < 0.01). In conclusion, the increasing MEC exfoliation rate in milk from mastitis udders caused by S. aureus may contribute to reduced milk yield.     

Relationship between resistance to vole browsing and content of ether extract in the bark of larch species and hybrids

We determined the survival rate of tree families produced by the intra- and interspecific crossing of larch species damaged by browsing of vole (Clethrionomys rufocanus bedfordiae Thomas) and the contents of ether extracts (EE) in their bark. The average survival ofLarix gmelini var.japonica Pilg. (L. gmelini)×L. gmelini (G×G) was highest, followed by the hybridL. gmelini×L. leptolepis Gordon (G×L). The families of the hybrid G×L, which is considered suitable for silviculture in Hokkaido, Japan, showed significant differences in survival rates, suggesting that it is possible to produce improved varieties of the hybrid which are more resistant to vole browsing. The EE contents of the bark were under strong hereditary control, and the rank of the EE content among the various families hardly fluctuated between the location of the test fields. The correlation coefficient between the survival rates and the EE contents was highly significant (r=0.89,n=24,p<0.01), indicating that the EE content in the bark may be used as a possible index of resistance or susceptibility to vole browsing in larch plantations. A part of this report was presented at the 106th Annual Meeting of the Japanese Forestry Society, at Sapporo, April, 1995.     

Presence of a Minor Amylose Fraction in the Central Portion of Waxy Wheat Starch Granules and Its Contribution to Granular Stability

When wheat starch granules containing various amounts of amylose (2.1–25%) were stained with 25% KI/10% I₂ solution, the granules largely changed to ghost structures below ≈5.0% amylose. The ghost showed a typical double structure: a black‐brown central portion and a red‐brown surrounding portion. The proportion of the black‐brown central portion in the ghost was strongly correlated to the amylose content (%) in the starch, that is, the black‐brown central portion decreased with a decrease in amylose. This suggests that amylose is possibly present in the black‐brown central portion. Sonication (20 kHz) followed by centrifugation of the ghost separated the black‐brown central portion from the red‐brown surrounding portion, and the amylose content in each portion was determined. The results indicated that the amylose content in the black‐brown central portion was 6.9%, whereas in the surrounding portion, it was only 1.0%. Furthermore, the central and surrounding portions were subjected to Sepharose CL‐2B gel‐filtration column chromatography and the presence of amylose could only be observed in the black‐brown central portion.     

Comparative anatomy of the radial sesamoid bone in the polar bear (Ursus maritimus), the brown bear (Ursus arctos) and the giant panda (Ailuropoda melanoleuca).

Since we have clarified the manipulation mechanism using the radial sesamoid (RS) in the giant panda (Ailuropoda melanoleuca), our aim in this study is to examine the position, shape and function of the RS morphologically, and to observe the attachment to the RS of the M. abductor pollicis longus and the M. opponens pollicis in the other Ursidae species. So, we focused on the carpus and manus of the polar bear (Ursus maritimus) and the brown bear (Ursus arctos) in this study. The RS was tightly articulated to the radial carpal, and could not adduct-abduct independently of the radial carpal. The M. abductor pollicis longus tendon and the M. opponens pollicis belly were attached to the RS, independently. In the polar bear, the deep concave and the flat surface were confirmed in attachment area for these two muscles. The morphological relationship between the RS and the M. abductor pollicis longus and the M. opponens pollicis in the two species of bears were essentially consistent with that in the giant panda. It also demonstrated that the manipulation mechanism of the giant panda has been completely based on the functional relationship between the small RS, and the M. abductor pollicis longus and the M. opponens pollicis in Ursidae species.     

CT examination of the guttural pouch (auditory tube diverticulum) in Przewalski's Horse (Equus przewalskii).

The domestic horse (Equus caballus) have the large symmetrical guttural pouches (the auditory tube diverticulum) formed by saccate bulge of the auditory tube. In this study, CT examination was carried out in the head of Przewalski's horse (Equus przewalskii), the only true wild horse living at present. As results of the examination, Przewalski's horse possessed the large symmetrical guttural pouches divided into medial and lateral compartments by the stylohyoid bone. Moreover, the right and left guttural pouches meet each other at the median part to form a thin septum. As CT sections get close to the part of the occipital condyle, the lateral compartment disappeared, and the medial compartment gradually became small toward the base of the skull. These results indicate that the nuchal-basal part of the medial compartment is not well-developed as compared with the domestic horse.     

Codend selectivity for jack mackerel and whitefin jack and unequal split parameter estimates observed in trouser trawl experiments

Codend selectivity for the jack mackerel Trachurus japonicus and the whitefin jack Kaiwarinus equula were evaluated based on data from trouser trawl experiments carried out in the East China Sea, using a test codend of 60 mm diamond mesh and a control codend made of minnow net with a square mesh of 9 mm bar length. Between-haul variations in parameters and the mean selection curves were tested with the catch data in the SELECT approach, and then the model of between-haul variation in the split parameter with the mean selection curve was chosen as the best fit using Akaike’s information criterion model selection. The 50% retention lengths and the selection ranges were 11.4 and 3.36 cm for jack mackerel and 8.83 and 0.93 cm for whitefin jack, respectively. The selection curve for whitefin jack was sharp, whereas that for jack mackerel was relatively wide. As the estimated split parameters indicated, about 80% of the whitefin jack entered the control codend, but 85 and 90% of the jack mackerel entered the control codend in the second and third hauls, respectively. The inequality in the split parameter is discussed from the viewpoint of the animal’s swimming behavior and water movement based on underwater video observations.     

Effect of cultivation methods on nutritional enrichment of euryhaline rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis>

This study aimed at comparing fatty acid contents of rotifers cultured with different methods after nutritional enrichment in order to evaluate the rotifer quality produced by these methods. Rotifers were cultured using either a batch or a continuous culture. From the batch culture, three experimental subpopulations were used, sampled from the culture at 1, 24, and 48 h after rotifer inoculation. The continuous culture was performed with two tanks; one was for cultivation with continuous feeding and water supply (cultivation tank), and another was for harvesting from the cultivation tank by overflow (harvest tank). From the continuous culture, two subpopulations were used: one from the cultivation and one from the harvest tank. Nutritional enrichment was performed after each culture. Each population was enriched with Nannochloropsis oculata or a commercial enrichment diet. When the enrichment was performed with N. oculata on populations at 24 h after inoculation originating from either of the two tanks of continuous culture or the batch culture tank, a higher quantity of arachidonic acid (ARA) and eicosapentaenoic acid (EPA) was obtained from the two tanks of continuous culture. The same results were obtained when enrichment diet was used, this time including docosahexaenoic acid (DHA).     

Amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis> interleukin-10 negatively suppresses host cell-mediated immunity

Interleukin-10 (IL-10) is an anti-inflammatory cytokine and negatively regulates cell-mediated immunity (CMI) induction by inhibiting cytokine production in type 1 T helper cells. IL-10 genes have been isolated from several fish, and inflammatory cytokine inhibition by IL-10 has been well examined. However, a CMI regulator of IL-10 in fish has not yet been identified. In this study, we cloned the IL-10 gene in amberjack Seriola dumerili and analyzed its function using its recombinant protein (rIL-10). In an in vitro culture experiment, gene expression of inflammatory cytokines was suppressed in leukocytes incubated with rIL-10 compared with cells that only received Nocardia seriolae stimulation. This result suggests amberjack IL-10 has conserved function as an inflammatory cytokine inhibitor. Bactericidal activity of amberjack cells against intracellular pathogen stimulation was decreased in a rIL-10 dose-dependent manner. Furthermore, a significant reduction in the T-bet/GATA-3 ratio was observed in N. seriolae living cell (LC)?+?rIL-10-injected fish. Taken together, these results suggest amberjack rIL-10 suppresses CMI induction both in vitro and in vivo. In addition, the number of IgM⁺ cells among spleen leukocytes in N. seriolae?+?rIL-10-injected fish was higher than in only N. seriolae LC, suggesting that Th2-dominant immunity was induced by adding rIL-10.