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Mycoplasma mastitis is a dairy herd health problem with growing concern in Japan. To complement the lack of epidemiological knowledge of the disease, we conducted estimation of herd-level prevalence and risk factor analysis for Mycoplasma bovis (M. bovis) mastitis using data collected from Tokachi region, one of the nation's largest milk producing area, in Hokkaido Prefecture in 2015. The herd-level prevalence was estimated at 3.8% (95% confidence interval (CI): 2.6%, 5.4%) across the region with municipality-specific prevalence ranging from 0% to 14.3%. In this study, identified risk factors for the presence of with M. bovis infection on farms were corporation-type farms and purchased cattle, after controlling for the herd size. Corporation-type farms may reflect higher frequencies of moving cows and fomites to and from other farms, which increases the risk of pathogen introduction. Purchased cattle was considered as one of the major pathways of the disease incursion, and this finding highlighted the importance of more stringent separation or quarantine protocols when introducing cattle from outside in Tokachi region. Due to the limited information available in this study and inherent nature of the study designs, these results should be interpreted with caution and further research is needed.

Concentrations of serum lipid components, lipid peroxide (LPO) and alpha-tocopherol and electrophoretic patterns of lipoproteins in serum samples obtained from captive marine mammals and terrestrial mammals were compared. Serum concentrations of total cholesterol, free fatty acid, and phospholipid in fish-eating animals were significantly higher than those in manatees and cows. Serum LPO and alpha-tocopherol concentrations in the fish-eating animals were also significantly higher than those in manatees, cows and dogs. Different patterns of densitometric scans of low density lipoprotein (LDL) and a significantly lower percentage of LDL were demonstrated in the dolphins compared with the seals, cow and dogs. The concentration of LPO was significantly correlated with triglyceride and phospholipid concentrations in serum from the dolphins. These results suggest that triglyceride and phospholipid are susceptible to oxidative reaction in fish-eating animals. Evaluation of serum lipids, LPO and alpha-tocopherol concentrations is needed for nutritional husbandry for fish-eating animals.

Haptoglobin cannot be detected in the serum of healthy cattle by the haemoglobin-binding assay or single radial immunodiffusion. The present study was designed to examine whether an enzyme-linked immunosorbent assay could be applied to measure serum haptoglobin concentrations in healthy cows and in cows with fatty liver. When either purified cow haptoglobin or haptoglobin-positive serum were used as the antigen in the assay, the standard curves obtained were distinctly different, and haptoglobin in serum was detected more sensitively than the purified protein. The addition of bovine serum albumin to purified haptoglobin shifted the curve towards that obtained with haptoglobin-positive serum, suggesting that an interaction with serum albumin was partly responsible for the different standard curves. By use of the standard curve for haptoglobin in serum, the mean (SEM) concentration of haptoglobin in samples from four cows with fatty liver was 466 (147) micrograms ml-1, but the sera from four apparently healthy cows contained less than 0.01 microgram ml-1 haptoglobin.

Biosynthesis of CD11/CD18 in bovine leucocytes, intracellular Ca 2+ ([Ca 2+ ] i ) signalling, chemiluminescent responses and membrane fluidity of neutrophils and the effects of D-mannose on neutrophils from control heifers and a heifer with bovine leucocyte adhesion deficiency ( BLAD ) were measured. The synthesis of CD11/CD18 complex was clearly detected in leucocytes from a normal heifer, but not in a BLAD -affected heifer. The transient phase of increased [Ca 2+ ] i was clearly detected in neutrophils from a heifer with BLAD stimulated with opsonised zymosan, aggregated bovine immunoglobulin G or concanavalin A, whereas the sustained phase was deficient or significantly decreased compared with control heifers. [Ca 2+ ] i signalling of neutrophils from control heifers and a heifer with BLAD stimulated with phorbol myristate acetate via an 11b/CD18-independent pathway showed no transient phase, and the subsequent increase in [Ca 2+ ] i was almost identical in neutrophils from affected and control heifers. [Ca 2+ ] i concentration and chemiluminescent responses of neutrophils from a control heifer were clearly decreased by treatment with anti-CD18 and anti-IgG antibodies. No differences in membrane fluidity were detected between neutrophils derived from control and CD18-deficient cattle. D-mannose binds mainly to Fc rather than CD18 receptors, and decreased Agg-IgG induced [Ca 2+ ] i and the chemiluminescent response of neutrophils. The [Ca 2+ ] i responses and Agg-IgG induced chemiluminescent responses of neutrophils from control heifers and a BLAD -affected heifer were inhibited by D-mannose. The characteristic changes of [Ca 2+ ] i signalling and functional responses of B 2 -integrin-deficient neutrophils were demonstrated.

Superoxide (O2) production and intracellular signal transduction of neutrophils were evaluated in five Holstein dairy calves and five lactating cows. Opsonised zymosan (OPZ)-induced O2 production by neutrophils from neonatal calves was significantly higher (P0.01) than that of neutrophils of cows, whereas heat-aggregated IgG (H-agg.IgG)- and phorbol myristate acetate (PMA)-induced O2 production of neutrophils were significantly lower (Por =0.01) than those of cows. To clarify the functional differences of intracellular signal transduction in neutrophils between neonatal calves and cows, the activities of protein kinase C (PKC) and tyrosine phosphorylation were evaluated in OPZ-, H-agg.IgG- and PMA-stimulated neutrophils. Membrane-associated PKC activity of OPZ-stimulated neutrophils from neonatal calves was significantly higher (P0.05) than that of cows, whereas PKC activity in membrane-associated fractions of H-agg.IgG-stimulated neutrophils from neonatal calves was significantly lower (P0.05) than that of cows. A significant difference was not found in membrane-associated PKC activity of neutrophils stimulated with PMA between neonatal calves and cows. The amount of tyrosine phosphorylated 100 kDa protein in neutrophils from neonatal calves stimulated with OPZ, H-agg.IgG and PMA were 192.6, 67.8 and 97.2 per cent of those of cows, respectively. These results indicate that complement receptor type 3 (CR3)- and Fc receptor (FcR)-mediated O2 producing activities of neutrophils are clearly different between neonatal calves and cows. This phenomenon may be associated with the age-related changes in intracellular signal transduction of neutrophils including PKC activity and tyrosine phosphorylation of cellular protein.