The effects of whole-plant corn silage (CS) particle size and long unprocessed grass hay (LH) supplementation on milk yield, chewing activity, and ruminal digestion in dairy cows were evaluated in 2 ...experiments. In Experiment 1, corn silage harvested at fine (6mm; FCS) or coarse (23mm; CCS) theoretical cut length were fed to 22 lactating Holstein cows. Treatments were 2 total mixed rations containing 58% of dry matter (DM) as FCS or CCS. Diet DM intake tended to be higher in cows fed FCS than those fed CCS (23.4 vs. 22.1kg/d). However, milk yield and composition, body condition score, and plasma metabolite concentrations were not affected by the dietary treatments. In the second experiment, 5 cannulated Holstein cows were used in a 5×5 Latin square design to evaluate the effects of the addition of LH to the diets evaluated in Experiment 1 on chewing activity and ruminal digestion. Treatments were 5 total mixed rations: FCS-based diet plus the addition of 0, 5, or 10% LH (DM basis) and CCS-based diet plus 0 or 5% LH. Long hay addition linearly decreased DM intake in cows fed FCS-based diets (25.0 to 21.7 kg/d), but increased DM intake in those fed CCS-based diets (22.7 to 27.1 kg/d). The intake of neutral detergent fiber (NDF) increased with LH addition in CCS-based diets (7.6 vs. 9.4 kg/d). Rumination time increased (16.8 to 21.0 min/kg of DM intake) when LH was added to FCS-based diets, but it decreased when included in CCS-based diets (18.8 vs. 12.9 min/kg of DM intake). Ruminal pH was higher (5.9 vs. 5.7) and lag-time for in situ NDF disappearance was shorter (3.5 vs. 8.7h) for cows fed CCS compared with cows fed FCS. The rate of NDF disappearance tended to be higher for the CCS-based diet with 5% LH than for the diet with 0% LH (2.0 vs. 4.4%/h), but solids passage rate was not affected by the treatments. These results suggest that addition of LH to FCS-based diets does not affect ruminal environment or digestion, but depressed DM intake. In contrast, addition of LH to CCS-based diets may improve ruminal NDF digestion, increasing DM intake by reducing filling effect and time needed for rumination.
In 2014, the European Society for Immune Deficiencies (ESID) revised the common variable immunodeficiency (CVID) diagnosis criteria by incorporating new clinical and biological markers. The new ...definition appeared more restrictive but had not yet been evaluated in a large cohort of patients.
The objective of this study was to evaluate the impact of this new definition in a large cohort of patients with primary hypogammaglobulinemia.
Evaluation of 3 different CVID definitions (ESID/Pan-American Group for Immunodeficiency PAGID 1999, ESID 2014, DEFI 2015) in 521 patients included in the French DEFI study with a diagnosis of primary hypogammaglobulinemia.
Using the ESID/PAGID 1999 definition, 351 patients were classified as CVID. The new ESID 2014 definition excluded 62 (18%) patients. Most of them (n = 56; 90%) had a less severe disease, whereas 6 (10%) presented with a severe disease with major T-cell defect. We propose different criteria (occurrence of opportunistic infection or very low naive CD4+ T-cell count) to define this population with severe T-cell defect. Sixty-two patients fulfilled these criteria, represented 20% of the initial CVID population but accounted for 77% of the deaths, with a 5-year overall survival of 67.6% (95% confidence interval, 51.0-79.6), and were considered as late onset combined immunodeficiency (LOCID).
The new ESID definition for CVID still fails to exclude a large number of patients with severe T-cell defect. We propose a new definition (DEFI 2015) that excluded more patients with a T-cell defect and consider these patients as LOCID. This population has a poor outcome and should be considered as a distinct group requiring specific care.
BACKGROUND: Chikungunya virus (CHIKV) has been responsible for large epidemic outbreaks causing fever, headache, rash and severe arthralgia. So far, no specific treatment or vaccine is available. As ...nucleic acid amplification can only be used during the viremic phase of the disease, serological tests like neutralization assays are necessary for CHIKV diagnosis and for determination of the immune status of a patient. Furthermore, neutralization assays represent a useful tool to validate the efficacy of potential vaccines. As CHIKV is a BSL3 agent, neutralization assays with infectious virus need to be performed under BSL3 conditions. Our aim was to develop a neutralization assay based on non-infectious virus replicon particles (VRPs). METHODS: VRPs were produced by cotransfecting baby hamster kidney-21 cells with a CHIKV replicon expressing Gaussia luciferase (Gluc) and two helper RNAs expressing the CHIKV capsid protein or the remaining structural proteins, respectively. The resulting single round infectious particles were used in CHIKV neutralization assays using secreted Gluc as readout. RESULTS: Upon cotransfection of a CHIKV replicon expressing Gluc and the helper RNAs VRPs could be produced efficiently under optimized conditions at 32°C. Infection with VRPs could be measured via Gluc secreted into the supernatant. The successful use of VRPs in CHIKV neutralization assays was demonstrated using a CHIKV neutralizing monoclonal antibody or sera from CHIKV infected patients. Comparison of VRP based neutralization assays in 24- versus 96-well format using different amounts of VRPs revealed that in the 96-well format a high multiplicity of infection is favored, while in the 24-well format reliable results are also obtained using lower infection rates. Comparison of different readout times revealed that evaluation of the neutralization assay is already possible at the same day of infection. CONCLUSIONS: A VRP based CHIKV neutralization assay using Gluc as readout represents a fast and useful method to determine CHIKV neutralizing antibodies without the need of using infectious CHIKV.
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