Experimental Yersinia enterocolitica placentitis in sheep.

A strain of Yersinia enterocolitica of O serogroup 6,30 isolated from the liver of an aborted ovine fetus was inoculated intravenously into a group of pregnant ewes at about 90 days gestation and produced placentitis with abortion or delivery of infected lambs about 50 days later. Y. enterocolitica of the same serogroup was recovered from the necrotic placental cotyledons and most other fetal tissues and could be isolated from vaginal discharges of the ewes for a least 2 weeks after abortion. Histological changes were consistent with an acute bacterial necrotizing placentitis and systemic infection of the fetus. Subsequent pregnancies in the ewes proceeded to term without evidence of infection.     

An improved ferritin assay for canine sera

An improved serum ferritin assay for canine serum has been developed. It uses two monoclonal antibodies in a sandwich arrangement. Serum ferritin can be determined on undiluted canine sera with this assay. The recovery of ferritin added to canine serum ranged from 98 to 106%, the within-assay coefficient of variability was 3.3 to 4.5%, and the assay-to-assay variability was 9.8 to 10.2%. Serum ferritin from 61 apparently healthy dogs had a geometric mean of 252 ng/ml, with a range of 80 ng/ml to 800 ng/ml.     

Phosphorylation by p38 MAPK as an alternative pathway for GSK3beta inactivation

Glycogen synthase kinase 3beta (GSK3beta) is involved in metabolism, neurodegeneration, and cancer. Inhibition of GSK3beta activity is the primary mechanism that regulates this widely expressed active kinase. Although the protein kinase Akt inhibits GSK3beta by phosphorylation at the N terminus, preventing Akt-mediated phosphorylation does not affect the cell-survival pathway activated through the GSK3beta substrate beta-catenin. Here, we show that p38 mitogen-activated protein kinase (MAPK) also inactivates GSK3beta by direct phosphorylation at its C terminus, and this inactivation can lead to an accumulation of beta-catenin. p38 MAPK-mediated phosphorylation of GSK3beta occurs primarily in the brain and thymocytes. Activation of beta-catenin-mediated signaling through GSK3beta inhibition provides a potential mechanism for p38 MAPK-mediated survival in specific tissues.     

Soybean meal alters autochthonous microbial populations, microvilli morphology and compromises intestinal enterocyte integrity of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout were fed either a diet containing fishmeal (FM) as the crude protein source or a diet containing 50% replacement with soybean meal (SBM) for 16 weeks. An enteritis-like effect was observed in the SBM group; villi, enterocytes and microvilli were noticeably damaged compared with the FM group. The posterior intestine microvilli of SBM-fed fish were significantly shorter and the anterior intestine microvilli significantly less dense than the FM-fed fish. Electron microscopy confirmed the presence of autochthonous bacterial populations associated with microvilli of both fish groups. Reduced density of microvilli consequently led to increased exposure of enterocyte tight junctions, which combined with necrotic enterocytes is likely to diminish the protective barrier of the intestinal epithelium. No significant differences in total viable counts of culturable microbial populations were found between the groups in any of the intestinal regions. A total of 1500 isolates were tentatively placed into groups or genera, according to standard methods. Subsequent partial 16S rRNA sequencing revealed species that have not been identified from the rainbow trout intestine previously. Compared with the FM group levels of Psychrobacter spp. and yeast were considerably higher in the SBM group; a reduction of Aeromonas spp. was also observed.     

Ferric pseudobactin 358 as an iron source for carnation

The influence of the siderophore produced by plant growth‐promoting Pseudomonas putida strain WCS358, pseudobactin 358 (PSB358), on chlorophyll synthesis and iron(III) reduction in carnation cultivars Lena and Pallas grown in hydroponics was studied. Ferric pseudobactin 358 (4 μM) stimulated chlorophyll synthesis in cv. Lena, but not in cv. Pallas. FeEDDHA stimulated chlorophyll synthesis in both cultivars. Differences between the two carnation cultivars in the utilization of FePSB358 as an iron source are correlated with differences in iron efficiency reactions of the two cultivars: Fe‐deficient plants of cv. Lena produced more and longer root hairs than Fe‐deficient plants of cv. Pallas, and the ferric reducing activity of cv. Lena was higher than that of cv. Pallas.     

First detection of 'Candidatus Mycoplasma haemolamae' infection in alpacas in England

This is the first report of detection of Candidatus Mycoplasma haemolamae in alpacas in England. The primary case occurred in a three year-old male alpaca in the south-east of England which presented with a history of progressive weight loss, lethargy, swelling of the scrotum and pale mucous membranes. Blood smear examination revealed a moderate, regenerative anaemia, with numerous small basophilic coccoid structures consistent with Candidatus M haemolamae. To confirm the presence of Candidatus M haemolamae, a portion of the 16S rDNA gene was amplified and analysed by denaturing gradient gel electrophoresis (DGGE). 16S rDNA gene sequencing showed a 99.8 per cent homology with Candidatus M haemolamae sequences deposited in GenBank. Subsequently, a cross-sectional study was carried out to investigate the presence of Candidatus M haemolamae infection in the alpaca herd from which the primary case was detected (n=131). Blood smear examinations and PCR with DGGE were used and compared with a species-specific PCR. The prevalence of infection when PCR positive results were combined was 29 per cent. A substantial agreement between the PCR/DGGE and the species-specific PCR was found (κ=0.86). A significant association was also found between age and infection (P=0.04) while no significant association was found with sex or origin.     

Evidence for functional G-coupled protein receptors 43 and 120 in subcutaneous and intramuscular adipose tissue of Angus crossbred steers

We conducted 3 independent experiments to demonstrate functional G-coupled protein receptor 43 (GPR43) and GPR120 in bovine intramuscular (i.m.) and subcutaneous (s.c.) adipose tissues. We hypothesized that media volatile fatty acids and long-chain fatty acids would affect cAMP-activated protein kinase-alpha (AMPKα) protein expression and cAMP concentrations differently in i.m. and s.c. adipose tissue. Experiment 1: oleic acid (18:1n-9) decreased phosphorylated AMPKα protein (p-AMPKα) and the p-AMPKα/AMPKα protein ratio in i.m. preadipocytes, increased the p-AMPKα/AMPKα protein ratio in bovine satellite cells, and had no effect in s.c. preadipocytes. Experment 2: ex vivo explants from the 5th to 8th longissimus thoracic rib muscle section of Angus crossbred steers were cultured 48 hr in media containing 0.25 µM ciglitizone, 5 mM glucose, and 5 mM acetate, in the absence or the presence of 100 µM oleic acid. Oleic acid increased acetate incorporation into fatty acids and GPR43 gene expression in i.m. adipose tissue (P < 0.05), but oleic acid had no effect on fatty acid synthesis or GPR43 expression in s.c. adipose tissue. Experiment 3: fresh s.c. and i.m. adipose tissue from the 5th to 8th longissimus thoracic rib muscle section of Angus crossbred steers was transferred immediately to 6-well culture plates containing 3 mL of KHB/Hepes/5 mM glucose. Samples were preincubated with 0.5 mM theophylline plus 10 μM forskolin for 30 min, after which increasing concentrations of acetate or propionate (0, 10⁻³, 10^−2.3, and 10⁻³ M) in the absence or the presence of 100 μM oleic acid or 100 µM palmitic acid (16:0) were added to the incubation media. Acetate had no effect on forskolin-stimulated cAMP production in s.c. adipose tissue but decreased cAMP in i.m. adipose tissue (P < 0.05); this indicates a functional GPR43 receptor in i.m. adipose tissue. The combination of 10⁻² M acetate and oleic acid decrease cAMP production in s.c. adipose tissue, consistent with GPR120 receptor activity, but oleic acid and palmitic acid attenuated the depression of cAMP production caused by acetate in i.m. adipose tissue. Palmitic acid depressed cAMP production in s.c. adipose tissue, and increased cAMP production in i.m. adipose tissue (P < 0.05). Propionate had no effect on cAMP production in s.c. or i.m. adipose tissue. These results provide evidence for functional GPR43 receptors in i.m. adipose tissue and GPR120 receptors in s.c. adipose tissue, both of which would suppress lipolysis.     

Kalmia removal increases nutrient supply and growth of black spruce seedlings: An effect fertilizer cannot emulate

In Canada's eastern boreal forest, the stagnant growth of black spruce (Picea mariana (Mill.) BSP) seedlings is often observed in the presence of ericaceous shrubs such as Kalmia angustifolia L. Many mechanisms, including allelopathic interference, reduced soil N mineralization, soil enzymes inhibition, and direct resource competition have been proposed to explain poor spruce growth in the presence of Kalmia. However, the relative importance of direct competition versus indirect interference remains unclear. Our objective was thus to adequately isolate the “Kalmia effect” from other growth-limiting factors and to determine if removal of Kalmia also resulted in fundamental changes in the biochemical properties of the forest floor. By sampling plots established in 2000, we evaluated how Kalmia eradication and spot fertilization influenced soil nutrient availability, N mineralization rates, microbial basal respiration and biomass, as well as planted black spruce seedling growth, dimensions, and foliar nutrient concentrations 6 years later. We measured higher extractable-P, mineralizable-N, seedling dimensions and growth rates, as well as lower extractable-K, total-K, basal respiration and microbial biomass, in plots without Kalmia than in those where Kalmia had been maintained from 2000 to 2006. Our results thus confirmed that Kalmia eradication over 6 years not only improved the growth and nutrition of black spruce seedlings, but also resulted in fundamental changes in the biochemical properties of the forest floor. We demonstrated that along with direct competition for resources, Kalmia interferes indirectly with black spruce by modifying nutrient cycling and energy fluxes in soil. Higher indices of available C in plots with Kalmia corroborates that Kalmia tannins or rhizodeposition may reduce N mineralization by stimulating microbial immobilization, a relation that however needs to be confirmed with longer term laboratory incubations. Our results indicated that although it had a positive influence on seedling growth, the fertilization effect was confined to the first few years following treatment application, and failed to influence soil processes as did Kalmia eradication. Further monitoring will indicate if the increased litterfall in fertilized plots will eventually initiate a second wave of fertilizer-induced changes to soil processes, as observed in other ecosystems.