Evaluation in mice of Brucella ovis attenuated mutants for use as live vaccines against B. ovis infection

Brucella ovis causes ram contagious epididymitis, a disease for which a specific vaccine is lacking. Attenuated Brucella melitensis Rev 1, used as vaccine against ovine and caprine brucellosis caused by B. melitensis, is also considered the best vaccine available for the prophylaxis of B. ovis infection, but its use for this purpose has serious drawbacks. In this work, two previously characterized B. ovis attenuated mutants (Δomp25d and Δomp22) were evaluated in mice, in comparison with B. melitensis Rev 1, as vaccines against B. ovis. Similarities, but also significant differences, were found regarding the immune response induced by the three vaccines. Mice vaccinated with the B. ovis mutants developed anti-B. ovis antibodies in serum of the IgG₁, IgG_2a and IgG_2b subclasses and their levels were higher than those observed in Rev 1-vaccinated mice. After an antigen stimulus with B. ovis cells, splenocytes obtained from all vaccinated mice secreted similar levels of TNF-α and IL12(p40) and remarkably high amounts of IFN-γ, a crucial cytokine in protective immunity against other Brucella species. By contrast, IL-1α -an enhancer of T cell responses to antigen- was present at higher levels in mice vaccinated with the B. ovis mutants, while IL-10, an anti-inflammatory cytokine, was significantly more abundant in Rev 1-vaccinated mice. Additionally, the B. ovis mutants showed appropriate persistence, limited splenomegaly and protective efficacy against B. ovis similar to that observed with B. melitensis Rev 1. These characteristics encourage their evaluation in the natural host as homologous vaccines for the specific prophylaxis of B. ovis infection.     

Neospora caninum infection as a cause of reproductive failure in a sheep flock

Neospora caninum has been detected only sporadically in cases of ovine abortion, and it has therefore traditionally been considered as an unimportant parasite in small ruminants. This study was carried out with the aim of identifying the pathogen causing serious reproductive problems on a commercial sheep farm. Sera from all rams and ewes tested negative for antibodies against Border disease virus, Schmallenberg virus and Coxiella burnetii, and infections by these agents were therefore ruled out. Nevertheless, seropositivity to N. caninum and/or Toxoplasma gondii was detected, although the seroprevalence was higher in the case of N. caninum. The percentage of lambings and the number of lambs per dam were significantly lower in ewes that were seropositive to N. caninum while no effect on these parameters was detected in ewes that were seropositive to T. gondii. There was also no evidence of infection by T. gondii in the foetal/lamb tissues analyzed by PCR and/or immunohistopathological techniques. On the contrary, the DNA of N. caninum was detected in 13 out of 14 foetuses/lambs descendant from dams seropositive to this parasite. Characteristic lesions caused by N. caninum and/or its antigen were also detected. Genotyping of the N. caninum DNA revealed only two closely related microsatellite multilocus genotypes. The results clearly demonstrate that infection by N. caninum was the cause of the low reproductive performance of this sheep flock.     

Molecular determination of the predator community of a cassava whitefly in Colombia: pest-specific primer development and field validation

In South America, the whitefly Aleurotrachelus socialis is one of the principal pests of cassava (Manihot esculenta Crantz), reaching high population levels throughout the Andean region. Management of this species is primarily based upon the use of insecticides, while biological control has received limited attention. Till present, knowledge of A. socialis natural enemies is restricted to occasional records of predators and parasitoids. In this study, we developed PCR primer sets specific for the cassava whitefly, A. socialis, to identify their predator community in Colombian cassava. Eleven percent of 586 predator specimens (representing 131 taxa from 29 families) tested positive for cassava whitefly DNA. Of the 21 predator taxa that consumed cassava whiteflies, an unidentified netwing beetle (Lycidae), an unidentified spider species (Araneae), Harmonia axyridis (Coleoptera: Coccinellidae), a Cereaochrysa sp. (Neuroptera: Chrysopidae), and a Leucochrysa sp. (Chrysopidae) were the taxa that consumed cassava whiteflies most frequently under field conditions. Two abundant predators in the system, Delphastus sp. (Coccinellidae) and the long-legged fly, Condylostylus sp. (Diptera: Dolichopodidae), were both positive for whitefly DNA, but did not have the strongest trophic linkage to the pest relative to other predators. This study shows that a diverse predator community affects cassava whitefly in southern Colombia, and provides the groundwork for the design of cassava production systems with minimal pesticide inputs.     

Shrubs facilitating seedling performance in ungulate-dominated systems: biotic versus abiotic mechanisms of plant facilitation

Shrubs are important microsites that facilitate seedling performance in abiotically stressed environments. However, shrub facilitation might work differently in ecosystems with high biotic stress (e.g., ungulate-dominated environments). We wonder whether ungulates through their preferences for different shrubs affect the facilitation process of seedlings, and whether this process remains consistent across different levels of abiotic stress. We study seedling performance (survival and growth) of a Mediterranean oak (Quercus pyrenaica) in three microsites (open, preferred and non-preferred shrubs by ungulates) for two contrasting climatic environments and throughout a complete growing season (increasing abiotic stress). Overall, there was a positive effect of shrubs in seedling survival. We found a microsite effect on seedling growth but this effect differed depending on the abiotic environment, with seedlings growing better in open microsites in the less stressful site. The larger abiotic context (climate) played an important role in determining which microsite is the most favorable for seedling growth but not necessarily for the eventual survival of seedlings. Wild ungulates started to damage seedlings early in the growing season, whereas abiotic stress (desiccation) affected mostly those plants that were not previously attacked by animals, showing the hierarchical effect (earlier and longer effect) of the biotic over the abiotic mortality agents. Non-preferred shrubs worked better as nurse shrubs only under high ungulate pressure. From the abiotic perspective, preferred and non-preferred shrubs did not apparently differ in the facilitation effect. Importantly, we found a greater facilitative effect of shrubs on the survival than on the growth of seedlings.     

The effects of long-day photoperiod on growth, body composition and skin colour in immature gilthead sea bream (Sparus aurata L.)

Immature gilthead sea bream (Sparus aurata L.) with a mean initial weight of 25.6 g were reared over 11 months to market size under different photoperiods: 16L:8D; 24L:0D and a control. Differences in final mean weight were significant between the three treatments (P<0.001): 16L:8D, 465.0 g; 24L:0D, 445.9 g; control, 402.6 g. Fish from the 24L:0D consumed most while 16L:8D fish converted it most efficiently. The lipid content of fillets was lower (P<0.05) for the 24L:0D (5.37±0.16 g 100 g^?1 of fillet) than for the control (6.02±0.12 g), and the moisture content lower for the control than for either treatment groups. Skin luminosity (L^*) was directly related to the number of hours of light exposure.     

Investigation of the Effects of Salinity and Dietary Protein Level on Growth and Survival of Pacific White Shrimp, Litopenaeus vannamei

It is presumed that in hypo‐ and hypersaline environments, shrimp’s requirements for some specific nutrients, such as protein, may differ from those known in the marine habitat; however, few investigations have been conducted in this area of study. In the present investigation, the effects of salinity and dietary protein level on the biological performance, tissue protein, and water content of Pacific white shrimp, Litopenaeus vannamei, were evaluated. In a 3 × 4 factorial experiment, juvenile shrimp with an average initial weight of 0.36 ± 0.02 g were exposed for 32 d to salinities of 2, 35, and 50 ppt and fed experimental diets with crude protein contents of 25, 30, 35, and 40%. A significant effect of salinity on growth of shrimp was detected, with the growth responses (final weight, weight gain) ranked in the order 2 ppt (3.87, 3.50 g) > 35 ppt (3.40, 3.04 g) > 50 ppt (2.84, 2.47 g). No effects of dietary protein level or an interaction between salinity and protein on growth of shrimp were observed under the experimental conditions of this study. Percent survival of shrimp fed the highest protein content (40%, survival of 74%) was, however, significantly lower than those of shrimp fed the other feeds (25, 30 and 35% protein, survival of 99, 91, and 94%, respectively), a result likely associated with the concentration of total ammonia nitrogen, which increased significantly at increasing protein levels. Final water content of whole shrimp was significantly lower in animals exposed to 50 ppt (70.8%) than in shrimp held at 2 (73.7%) and 35 ppt (72.3%). No effect of salinity, protein, or their interaction was observed on the protein content of whole shrimp. The results of the present study are in agreement with reports of superior and inferior growth of L. vannamei reared in hypo‐ and hypersaline environments, respectively, as compared to what is generally observed in seawater.     

Evaluating the role of ecosystem services in participatory land use planning: proposing a balanced score card

The application of the ecosystem services (ES) concept in land use planning has great potential to enhance the awareness of planning actors on their interactions. At the same time it can contribute to improve the linkage between the role of land use patterns and the understanding of land system functioning and its contribution to human well-being. The concept should be developed in a way that can be applicable in socio-ecological systems where nature and society are capable of enhancing their roles mutually. The objective of this paper is to suggest a standardized scheme and generalizable criteria to assess how successful the application of the ES concept contributed to facilitate participatory planning. We consider three potential advantages and three critical aspects for how to improve the applicability and relevance of the ES concept in planning. Hereon based, we present a balanced score card tool for which we broke down to advantages and risks into concrete questions. We illustrate the application of this approach with two case studies, representatives of two major governance schemes in relation to land use planning. We demonstrate that the balanced score card approach helps to reveal potential imbalances regarding the consideration of different ES groups. It supports testing the potential of the ES concept to enhance or not interactions of local and regional actors. We conclude that the framework should be reconsidered after a set of case studies to be developed into a monitoring tool for supporting planning practices.     

COMPARISON BETWEEN COMPUTED TOMOGRAPHY,FLUOROSCOPY, AND ULTRASONOGRAPHY FOR GUIDING PERCUTANEOUS INJECTION OF THE CANINE INTERVERTEBRAL DISC

A minimally invasive method for delivering injectable therapeutic agents would be desirable for the treatment of intervertebral disc disease in dogs. The purpose of this study was to compare computed tomography (CT), ultrasonography (US), and fluoroscopy modalities for guiding percutaneous injection into canine intervertebral discs. Intervertebral discs of 14 dog cadavers were injected with a gelified ethanol therapeutic agent. Successful injectate placement and injectate leakage were determined based on necropsy inspection of discs. Injection into the nucleus pulposus was successful in 55 of 78 (71%) of all injected discs. Injections guided using CT and fluoroscopy were significantly more successful than US‐guided injections. Odds of successful injection without leakage were greater for CT vs. US (P = 0.0026) but there was no significant difference between CT and fluoroscopy (P = 0.0620). Injection success rates did not differ among vertebral sites or dog cadavers of varying weights. Forty‐nine (63%) of injection sites had injectate leakage outside the disc and 10 of these involved structures within the vertebral canal. The highest rate of injection success with the least amount of leakage was achieved with CT guidance. Findings indicated that CT, fluoroscopy, and US are feasible modalities for guiding percutaneous injection of a gelified ethanol therapeutic agent into the canine intervertebral disc, with moderate to high success rates for different regions of the spine. However, a moderately high rate of injectate leakage occurred outside of the disc and this should be taken into consideration for future safety and efficacy studies.     

Recovery from neuromuscular block in dogs: restoration of spontaneous ventilation does not exclude residual blockade

ObjectiveTo evaluate if return of spontaneous ventilation to pre-relaxation values indicates complete recovery from neuromuscular blockade.Study designProspective, with each individual acting as its own control.AnimalsTen healthy adult female Beagle dogs weighing 6.2–9.4 kg.MethodsDogs were anesthetized with propofol, dexemedetomidine and isoflurane. Spontaneous ventilation was assessed by measuring end-tidal CO₂, expired tidal volume, peak inspiratory flow, respiratory rate and minute ventilation. Vecuronium 25 μg kg^?1 IV was administered and neuromuscular block was evaluated by measuring the train-of-four (TOF) ratio with acceleromyography in the hind limb. During spontaneous recovery from neuromuscular block, the TOF ratio when each ventilatory variable returned to baseline was recorded.ResultsThis dose of vecuronium produced moderate neuromuscular block in all dogs, with TOF ratio values of 0–18% at maximal block. Expired tidal volume, peak inspiratory flow and minute ventilation returned to pre-relaxation values when the median TOF ratio was ≤ 20%. The median TOF ratio was 42% when the end-tidal CO₂ returned to pre-relaxation values.Conclusions and clinical relevanceSignificant residual neuromuscular block could be measured at the hind limb with acceleromyography when ventilation had spontaneously returned to pre-vecuronium values. Monitoring spontaneous ventilation, including end-tidal CO₂, expired tidal volume, peak inspiratory flow or minute ventilation cannot be used as a surrogate for objective neuromuscular monitoring, and this practice may increase the risk of postoperative residual paralysis.     

Haplotype phasing after joint estimation of recombination and linkage disequilibrium in breeding populations

A novel method for haplotype phasing in families after joint estimation of recombination fraction and linkage disequilibrium is developed. Results from Monte Carlo computer simulations show that the newly developed E.M. algorithm is accurate if true recombination fraction is 0 even for single families of relatively small sizes. Estimates of recombination fraction and linkage disequilibrium were 0.00 （SD 0.00） and 0.19 （SD 0.03） for simulated recombination fraction and linkage disequilibrium of 0.00 and 0.20, respectively. A genome fragmentation phasing strategy was developed and used for phasing haplotypes in a sire and 36 progeny using the 50 k Illumina BeadChip by： a） estimation of the recombination fraction and LD in consecutive SNPs using family information, b） linkage analyses between fragments, c） phasing of haplotypes in parents and progeny and in following generations. Homozygous SNPs in progeny allowed determination of paternal fragment inheritance, and deduction of SNP sequence information of haplotypes from dams. The strategy also allowed detection of genotyping errors. A total of 613 recombination events were detected after linkage analysis was carried out between fragments. Hot and cold spots were identified at the individual （sire level）. SNPs for which the sire and calf were heterozygotes became informative （over 90%） after the phasing of haplotypes. Average of regions of identity between half-sibs when comparing its maternal inherited haplotypes （with at least 20 SNP） in common was 0.11 with a maximum of 0.29 and a minimum of 0.05. A Monte-Carlo simulation of BTA1 with the same linkage disequilibrium structure and genetic linkage as the cattle family yielded a 99.98 and 99.94% of correct phases for informative SNPs in sire and calves, respectively.