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.     

Resolvin D1, a Metabolite of Omega-3 Polyunsaturated Fatty Acid,Decreases Post-Myocardial Infarct Depression

We hypothesized that inflammation induced by myocardial ischemia plays a central role in depression-like behavior after myocardial infarction (MI). Several experimental approaches that reduce inflammation also result in attenuation of depressive symptoms. We have demonstrated that Resolvin D1 (RvD1), a metabolite of omega-3 polyunsaturated fatty acids (PUFA) derived from docosahexaenoic acid, diminishes infarct size and neutrophil accumulation in the ischemic myocardium. The aim of this study is to determine if a single RvD1 injection could alleviate depressive symptoms in a rat model of MI. MI was induced in rats by occlusion of the left anterior descending coronary artery for 40 min. Five minutes before ischemia or after reperfusion, 0.1 μg of RvD1 or vehicle was injected in the left ventricle cavity. Fourteen days after MI, behavioral tests (forced swim test and socialization) were conducted to evaluate depression-like symptoms. RvD1 reduced infarct size in the treated vs. the vehicle group. Animals receiving RvD1 also showed better performance in the forced swim and social interaction tests vs. vehicle controls. These results indicate that a single RvD1 dose, given 5 min before occlusion or 5 min after the onset of reperfusion, decreases infarct size and attenuates depression-like symptoms.     

The physiological tolerance of the grey carpet shark (<Emphasis Type="Italic">Chiloscyllium punctatum</Emphasis>) and the epaulette shark (<Emphasis Type="Italic">Hemiscyllium ocellatum</Emphasis>) to anoxic exposure at three seasonal temperatures

The epaulette shark (Hemiscyllium ocellatum) and the grey carpet shark (Chiloscyllium punctatum) are commonly found in periodically hypoxic environments. The ecophysiological time available for these animals to safely exploit these niches during different seasonal temperatures was examined. The time to loss of righting reflex (T _LRR) was examined in response to an open ended anoxic challenge at three seasonal temperatures (23, 25 and 27°C). Ventilation rates were measured in an open ended anoxic challenge at 23°C and during 1.5 h of anoxia followed by 2 h of re-oxygenation at 23 and 25°C. The mean T _LRR of epaulette and grey carpet sharks was inversely proportional to temperature. The T _LRR was similar between species at 23°C; however, grey carpet sharks had significantly reduced T _LRR at higher temperatures. During the standardised anoxic challenge, epaulette sharks entered into ventilatory depression significantly earlier at 25°C. During re-oxygenation, epaulette sharks exposed to anoxia at 23°C had no significant increase in ventilation rates. However, after anoxic challenge and re-oxygenation at 25°C, epaulette sharks showed a significant increase in ventilation rates during re-oxygenation. Grey carpet sharks displayed no evidence of ventilatory depression during anoxia. However, during re-oxygenation, grey carpet sharks had significantly elevated ventilation rates above pre-experimental levels and control animals. These data demonstrate that the anoxia tolerance times of both species were temperature dependent, with a significant reduction in the T _LRR occurring at higher temperatures. Epaulette sharks had a significantly greater T _LRR at higher temperatures than grey carpet sharks, which did not enter into a ventilatory depression.     

Microbiological Quality Assessment of Watershed Associated with Animal-Based Agriculture in Santa Catarina, Brazil

Environmental problems many times could evolve when manure-containing pathogens are distributed into an open environment with no effort made to reduce the content of pathogens or limit their movement in the environment. Wind, surface flow, and subsurface flow can all carry enough pathogens to receiving waters to exceed water quality standards. This study was conducted to assess the microbiological quality of water associated with animal-based agriculture in the sub-basin of Pinhal River located in the rural area of Concordia, Santa Catarina, Brazil. Six sampling points representing different agricultural land uses (LU1—dairy cattle; LU2—without animals; LU3—dairy + pigs + poultry + crops; LU4—pigs + poultry + crops; LU5—dairy + pigs + poultry + crops + human; and LU6—dairy+pigs+crops) along the Pinhal River sub-basin (north to south) were sampled biweekly from August 2006 to December 2008. Concentrations of fecal coliforms and Escherichia coli varied significantly (p?≤?0.05) with land use (LU), but there was no interaction effect of LU, season, and time. Water samples from the catchment area of LU1 had the highest concentration of fecal coliforms (4,479?±?597 CFU ml^?1) when compared with other catchment areas. Catchment area associated with LU2 (no animal) had the lowest concentrations of fecal coliforms (39.2?±?5.2 CFU ml^?1). With the exception of LU2 (control site), all the maximum concentrations of E. coli exceeded the single maximum allowable concentration for E. coli (100 CFU ml^?1). When LU1 was compared with other catchment areas (LU3, 50%; LU4, 67%; LU5, 58%; and LU6, 44%), it had the lowest counts (39%) of Salmonella sp. Our results suggest that spatial pattern of bacterial water quality is evident, which can be linked to the different land uses and associated practices (present or absent of animal activities). Therefore, varying responses associated with the different land uses would be critical in identifying the importance of different sources of bacteria in the catchment area and the mechanisms transferring them.     

The Functional Anatomy of the Neck and Forelimbs of the Cape Golden Mole,Chrysochloris Asiatica (Lipotyphla: Chrysochloridae)

The anatomy of the neck and forelimb of Chrysochloris asiatica is described and illustrated. The sequence of events during digging and modifications for fossorial action are described. Modifications include the appearance of a third bone in the forearm; the shortening and fusion of bones in the manus; enlarged processes on the scapula, humerus and ulna for greater muscle attachment; enlarged neck muscles and a dip in the spine in the cervical region to accommodate these and the enlarged shoulder muscles; an enlarged occiput for insertion of the powerful neck muscles; a greatly enlarged triceps and movement of the shoulder girdle to a position anterior to that normal in mammals. The possibility of the third forearm bone being the ossified tendon of a flexor muscle is discussed, without any conclusion being reached as to its true origin.     

Toxicokinetics of the active doxorubicin metabolite, doxorubicinol, in sulphur-crested cockatoos (Cacatua galerita)

The pharmacokinetics of doxorubicinol, a cytotoxic metabolite of the anticancer drug, doxorubicin, were studied in four healthy sulphur-crested cockatoos (Cacatua galerita) after a 20 min intravenous infusion of 2 mg/kg. Plasma doxorubicinol concentrations were measured by HPLC. The pharmacokinetic parameters were estimated using a non-compartmental method. The mean (+/- SD) peak concentration was 8341 +/- 3132 microg/L at 17.5 +/- 5.0 min after the start of the infusion, and doxorubicinol concentrations declined biexponentially to 154.3 +/- 34.5 microg/L, 40 min after the end of the infusion. Systemic clearance was 0.940 +/- 0.473 L/h/kg, mean residence time was 0.165 +/- 0.133 h, and steady-state volume of distribution was 0.123 +/- 0.0526 L/kg. The terminal half-life was 0.660 +/- 0.611 h. Detectible but unquantifiable concentrations of doxorubicinol were present in the plasma ultrafiltrate of two birds during the infusion, indicating very extensive plasma protein binding. Physiological, haematological and biochemical monitoring over 3 weeks showed that doxorubicinol at a single infused dose of 2 mg/kg caused no toxicities of major concern.     

Induction of an alloplasmic male-sterile Brassica oleracea by substituting cytoplasm from ‘Early Scarlet Globe’ radish (Raphanus sativus)

Summary Alloplasmic male-sterile Brassica oleracea L. was synthesized in a backcrossing program through amphidiploid Raphanobrassica by using Early Scarlet Globe radish (Raphanus sativus L.) as the donor of cytoplasm and B. oleracea broccoli and cabbage as recurrent pollen parents. Persistence of radish chromosomes and high female sterility were encountered in the first four backcrosses. Following use of colchiploid 4x broccoli as pollen parent, a BC₅ plant was obtained that had 2n=3x+1=28 chromosomes, improved seed set, and no radish traits. The BC₆ with recurrent 2x broccoli contained male-sterile plants with 2n=18 or 19 chromosomes, increased seed set, and broccoli morphology. Subsequent generations segregated for male-sterile and restored male-fertile plants, some with variable development of stamens and pollen. Leaf color of the alloplasmic plants, especially seedlings, was lighter green than normal.     

Chromosome behavior and sterility of hybrids between the common onion,Allium cepa,and the related wild A. oschaninii

Summary Four hybrid plants were obtained when cytoplasmic male-sterile onion inbreds, Allium cepa L., were pollinated with a wild cepa-like allium introduced from northeastern Iran and identified as A. oschaninii O. Fedtsch. It has been proposed on morphological grounds that A. oschaninii is wild A. cepa or the ancestor of A. cepa. Nevertheless, chromosome irregularities and complete sterility in the hybrids suggest that there may be a greater sterility barrier between A. cepa and A. oschaninii than between A. cepa and 3 other species reported to cross with A. cepa, namely A. fistulosum L., A. galanthum Kar. et Kir., and A. pskemense B. Fedtsch.Cooperative investigation of the U.S. Department of Agriculture and the Idaho Agricultura Experiment Station, Parma Branch Station.     

Continuous Plant Regeneration from Established Embryogenic Cell Suspension Cultures of Italian Ryegrass and Tall Fescue

The suitability of different protocols was compared for entire plant regeneration by somatic embryogenesis, of the forage plants Lolium multiflorum Lam. (Italian ryegrass) and Festuca arundinacea Schreb. (tall fescue). In the first protocol, miniature embryos were used as starting material, while mature seeds were retained in the other two. Whichever the considered protocol, undifferentiated calli were produced on Murashige and Skoog MS medium supplemented with 2,4-D. The calli were subcultured in the dark on solid MS agar medium, containing 5 mg/1 2,4-D (protocol 2) or on solid MS medium followed by transfer to a rotated liquid MS medium with 2 mg/1 2,4-D (protocol 1). In these conditions, induction of somatic embryogenesis occurred, and whole plants were regenerated during a limited lapse of time, upon transfer in the light, to MS medium supplemented with BAP but devoid of 2,4-D. The simultaneous elimination of 2,4-D and transfer to light appeared essential for full regeneration of the plants. Using this characteristic, an additional step was added to a new protocol (protocol 3) in which microcalli, cultured on liquid MS medium containing 5 mg/1 2,4-D, were transferred to the same medium with 2 mg/1 2,4-D, in the dark. In these conditions, the suspensions kept their embryogenic potential for months. In all cases, plantlets were successfully transferred into the soil. An evaluation of the somaclonal variation potential of the plants issued from each protocol is now underway.