Immunity,safety and protection of an Adenovirus 5 prime - Modified Vaccinia virus Ankara boost subunit vaccine against Mycobacterium avium subspecies paratuberculosis infection in calves

Vaccination is the most cost effective control measure for Johne’s disease caused by Mycobacterium avium subspecies paratuberculosis (MAP) but currently available whole cell killed formulations have limited efficacy and are incompatible with the diagnosis of bovine tuberculosis by tuberculin skin test. We have evaluated the utility of a viral delivery regimen of non-replicative human Adenovirus 5 and Modified Vaccinia virus Ankara recombinant for early entry MAP specific antigens (HAV) to show protection against challenge in a calf model and extensively screened for differential immunological markers associated with protection. We have shown that HAV vaccination was well tolerated, could be detected using a differentiation of infected and vaccinated animals (DIVA) test, showed no cross-reactivity with tuberculin and provided a degree of protection against challenge evidenced by a lack of faecal shedding in vaccinated animals that persisted throughout the 7 month infection period. Calves given HAV vaccination had significant priming and boosting of MAP derived antigen (PPD-J) specific CD4⁺, CD8⁺ IFN-γ producing T-cell populations and, upon challenge, developed early specific Th17 related immune responses, enhanced IFN-γ responses and retained a high MAP killing capacity in blood. During later phases post MAP challenge, PPD-J antigen specific IFN-γ and Th17 responses in HAV vaccinated animals corresponded with improvements in peripheral bacteraemia. By contrast a lack of IFN-γ, induction of FoxP3+ T cells and increased IL-1β and IL-10 secretion were indicative of progressive infection in Sham vaccinated animals. We conclude that HAV vaccination shows excellent promise as a new tool for improving control of MAP infection in cattle.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0112-9) contains supplementary material, which is available to authorized users.     

Restoring silvopastures with oak saplings: effects of mulch and diameter class on survival,growth, and annual leaf-nutrient patterns

In Southwestern Spain, multifunctional silvopastoral systems consisting of pastureland and open oak woodlands are known as Dehesas. These, and other similar systems of the Mediterranean basin, are currently threatened by increasing intensive land use. As a consequence, oak regeneration is declining and is in need of adequate management and active restoration. Traditional restoration practices outplant one-year-old, nursery-produced oak seedlings grown in 250–350 cm³ containers, but establishment and growth results are typically poor. This work examines holm oak (Quercus ilex L. subsp. ballota (Desf.) Samp.) grown in a non-conventional container size (24 l) and age (6–7 years) with three mulch treatments (control, stone, and straw). In an open Dehesa of SE Spain, 106 oak saplings were planted in March 2010, and survival, diameter at breast height (DBH), and leaf-nutrient concentrations were analyzed. Forty months after planting, all treatments showed high survival (81 %) but only straw-mulched saplings differed significantly (94 %) from control (74 %). DBH increased over time but showed no significant differences among mulch treatments. Saplings with high initial DBH showed the greatest growth and change in DBH at the end of the study period. Leaf-nutrient concentrations changed significantly in the year following outplanting. Bi-monthly foliar nutrient concentration data show decreases in P, K, Zn, and B and sharp increases in Ca and Fe. In this work, we provide some evidence concerning the viability of non-conventional oak size for restoring, regenerating, or building up new agroforestry or silvopastoral systems. A combination of saplings with more than 10 mm of DBH and straw mulch is recommended.     

Preliminary characterization and interaction of tubulin from Trichinella spiralis larvae with benzimidazole derivatives

Tubulin was estimated to account for 0.3% of the total soluble protein in Trichinella spiralis cytosolic fractions. Tubulin from T. spiralis was partially purified by precipitation with either taxol or vinblastine sulphate. Immunoblotting with alpha- and beta-tubulin monoclonal antibodies revealed the presence of tubulin in T. spiralis partially purified preparations. Electrophoretic mobility of T. spiralis tubulin in sodium dodecyl sulphate-polyacrylamide gels was very similar to that shown by pig brain tubulin. Further studies with colchicine binding assays indicated that T. spiralis tubulin has binding features similar to that of tubulin from other nematodes: colchicine association constant = 8.1 x 10(-4) M and competitive inhibition of colchicine binding by podophyllotoxin, with an inhibition constant of 1.3 x 10(-6) M. Finally, inhibition of colchicine binding by several benzimidazoles (mebendazole, fenbendazole, oxibendazole and albendazole) was investigated. All the benzimidazoles inhibited colchicine binding in a competitive manner, with inhibition constant values ranging from 1.4 x 10(-7) M (mebendazole) to 3.9 x 10(-6) M (fenbendazole).     

猪的回肠炎及其控制

猪的增生性回肠病 (回肠炎 )最早在 1 931年被发现 ,但直到约 60年后 ,本病的病原体由爱丁堡大学的 lawson等人命名为 lawsonia intracellu-laris。随后的 1 0年中由于许多特有技术的发展 ,使得人们可进一步研究这种细菌感染的流行病学和它的各种临床表现 -即地方性回肠炎 ( RI)、增生性肠病 ( PE)、坏死性肠炎 ( NE)和急性出血增生性肠病 ( AHPE)。所有疾病的控制方案都是以流行病学为基础的。有关病原体和传播方式的许多问题仍待解决。1　流行病学分子诊断技术可用于整个畜群抽查并可检测它们粪便中的 L intracellularis。西班牙对 7…     

Immune responses of piglets to weaning stress: impacts of photoperiod

An acute stress response can be provoked by abrupt social, nutritional, and environmental changes associated with weaning, and this may disrupt homeostasis and thus compromise well-being. Manipulating environmental factors, such as photoperiod, might provide a simple way to reduce the physiological consequences that piglets experience due to weaning stress. The objective of this study was to evaluate the impacts of photoperiod manipulation across various weaning ages on leukocyte populations, lymphocyte proliferation, natural killer cytotoxicity (NK), chemotaxis, phagocytosis, and immunoglobulin G, cortisol, and BW of piglets during the nursery phase. Sixty-eight crossbred piglets were obtained from sows kept on a short-day (8 h of light/d) photoperiod from d 90 of gestation until weaning. Piglets were weaned at 14, 21, or 28 d of age and kept on a short or long (16 h of light/d) photoperiod until 10 wk of age. Piglet BW and blood samples were collected at weaning and at 6, 8, and 10 wk of age. Pigs weaned at 28 d had reduced neutrophil counts (P < 0.001), phagocytosis (P < 0.001), and lymphocyte proliferation (P < 0.05) at weaning compared with those weaned at 14 and 21 d. Pigs weaned at 21 d tended to have lower (P = 0.08) lymphocyte counts than did pigs weaned at 14 or 28 d. Pigs weaned at 14 d had reduced (P < 0.01) NK relative to those weaned at 21 or 28 d. Photoperiod also influenced pig BW and immune status. Generally, those pigs on the long-day photoperiod and weaned at 28 d were heavier (P < 0.001) than their counterparts weaned at 14 or 21 d. At 6 wk of age, NK was greater (P = 0.002) in pigs kept on a long day and weaned at 14 or 21 d than in pigs weaned at 28 d. Phagocytosis was less (P = 0.005) at 6 wk of age, but was greater at 8 wk, in piglets kept on the long day and weaned at 28 d than in long-day pigs weaned at 14 or 21 d. These results suggest that photoperiod differentially influences immune responses in piglets weaned at different ages and indicate an inverse relationship between growth and immune status. Here, weaning at 28 d and a long-day photoperiod was the treatment combination that was most physiologically beneficial to piglets, whereas a 14-d weaning and short-day photoperiod was least physiologically beneficial.     

Validation of candidate bovine reference genes for use with real-time PCR

Accurate quantification with real-time PCR requires the use of stable endogenous controls. Recently, there has been much debate concerning the stability of commonly used reference or housekeeping genes. To address this concern, a number of statistical approaches have been designed to analyse data and assist in determining the most appropriate reference genes for experimental comparisons. In this study, three programs, BestKeeper, Norm Finder, and geNorm were used to assess four candidate reference genes: 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), acidic ribosomal protein large (RPLP0) and beta-actin, for use in expression profiling of individuals from divergent cattle genotypes subject to parasitic challenge with the cattle tick Boophilus microplus. Results demonstrated beta-actin and GAPDH were the most suitable reference genes in blood and could be used either individually or combined as an index to normalise data. RPLP0 was identified as the least stable gene, while 18S rRNA was omitted as being too highly expressed. As the recommendations on the most suitable reference genes varied between the programs, it is recommended that more than one should be utilised, to ensure the most robust experimental tools are selected.