Genetic conservation of potentially immunogenic proteins among Brazilian isolates of Babesia bovis

Bovine babesiosis caused by Babesia bovis remains an important constraint for the development of cattle industries worldwide. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these vaccines have a number of drawbacks, which justifies the search for better, safer vaccines. In recent years, a number of parasite proteins with immunogenic potential have been discovered. However, there is little information on the genetic conservation of these proteins among different parasite isolates, which hinders their assessment as immunogens. The aim of the present study was to evaluate the conservation of the genes ama-1, acs-1, rap-1, trap, p0 and msa2c among five Brazilian isolates of B. bovis. Through polymerase chain reaction, genetic sequencing and bioinformatics analysis of the genes, a high degree of conservation (98-100%) was found among Brazilian isolates of B. bovis and the T2Bo isolate. Thus, these genes are worth considering as viable candidates to be included in a recombinant cocktail vaccine for cattle babesiosis caused by B. bovis.     

Molecular epidemiology of foot-and-mouth disease virus type A in South America

A databank of 78 VP(1) complete sequences of type A foot-and-mouth disease virus (FMDV) from South American isolates was constructed. Forty-nine samples corresponded to FMDV that circulated between the years 1999-2008, mainly in Venezuela, where most type A outbreaks have occurred lately and twenty-nine to strains historically relevant for the continent. The phylogenetic analysis showed that all South American FMDV belonged to the Euro-SA topotype. Sixteen subgenotypes could be identified, based on a 15% nucleotide divergence cut-off criterion: eight are extinguished, three were active until the year 2002 and the remaining five circulated in Venezuela during the years 2001-2007, illustrating the potential for FMDV diversification under appropriate selective pressure. The last emergencies reported in already-free areas of Colombia in 2004 and 2008 were closely related to isolates acting in Venzuela. Evidence of positive selection over codon 170, within the immunogenic site 4 of VP1 protein, was recorded. A codon deletion in amino acid position 142, within the G-H loop, was found in some isolates within subgenotypes 14, 15 and 16. Conversely amino acid deletion 197 was restricted to all isolates within a particular genetic cluster. The present work is the first comprehensive phylogenetic analysis of FMDV type A in South America, filling a gap of knowledge with respect to both, historical and acting viruses. The results provided evidence that supports the ecosystem dynamics in the region, and also served as an input to establish genetic links of emergencies in already-declared free areas, highlighting the need for strengthening control activities.     

Assessing soil carbon lability by near infrared spectroscopy and NaOCl oxidation

The feasibility of near infrared (NIR) spectroscopy for quantifying labile organic matter (OM) in arable soils and for predicting soil refractory OM fractions was tested on 37 soils varying in texture and soil carbon (C) content. Three sets of arable soils (0-20 cm depth) were sampled from 1) long-term field experiments with different OM inputs, 2) individual sites with inherent with-in field gradients in soil texture and/or C content, and 3) from a range of different sites covering variations in management and geological origin. The labile OM fraction was defined by the CO₂ evolved from the soils incubated for 34 weeks while refractory OM was obtained by NaOCl oxidation.The labile fraction of the soil C accounted for 2-12% of the total soil C content. No systematic relationship between labile C content and total soil C or clay was found, but NIR spectra could be correlated well with the labile C fraction. A distinct, close linear relationship was found for C in soil before and after the NaOCl oxidation, indicating that this method was unable to reveal any additional information not contained in the total soil C measurement. NIR was also correlated with the NaOCl resistant C fraction, but this was considered to relate to the ability of NIR to predict total soil C contents. Thus NIR seemed to have the potential to estimate labile OM determined under laboratory incubations, while it still remains open how to identify and quantify refractory pools of soil OM.     

Organic carbon in soils of Germany: Status quo and the need for new data to evaluate potentials and trends of soil carbon sequestration

Uncertainties in estimates of soil carbon (C) stocks and sequestration result from major gaps in knowledge of C storage in soils, land‐use history, the variability of field measurements, and different analytical approaches applied. In addition, there is a lack of long‐term datasets from relevant land‐use systems. As in many European countries, a national database on soil organic carbon (SOC) including all relevant information for the determination of soil C stocks is likewise missing in Germany. In this paper, we summarize and evaluate the present state of knowledge on organic‐C contents/pools in soils of Germany and discuss the need for the acquisition and access to new data on soil organic carbon. Despite the number of agricultural sites under permanent soil monitoring, regional surveys on SOC, comprehensive ecosystem studies, and long‐term field experiments, there is a striking lack of data in Germany particularly with regard to agricultural soils. Apart from a missing standardization of methods and homogeneous baseline values, the implementation of a periodic, nation‐wide soil inventory on agricultural soils is required in order to simultaneously record information on land use, land‐use change, and agricultural practice. In contrast, the existing national inventory of forest soils provides information on C‐stock changes in forest soils, although there is some concern with regard to the representativeness of the sampling design to adequately address the problem of spatial heterogeneity and temporal variability. It is concluded that the lack of comprehensiveness, completeness, actuality, data harmonization, and standardized sampling procedures will further prevent the establishment of a SOC database in Germany with regard to the monitoring of trends in soil C pools and fluxes and the assessment of long‐term C‐sequestration potentials of soils under different land use. A future soil inventory should represent the heterogeneity of organic matter through functionally different SOC pools, topsoil characteristics as well as content, pool, and flux data for the deeper mineral‐soil compartments.     

Hemoglobin-mediated lipid oxidation and compositional characteristics of washed fish mince model systems made from cod (Gadus morhua), herring (Clupea harengus), and salmon (Salmo salar) muscle

The use of washed cod light muscle minces in mechanistic studies of hemoglobin (Hb)-mediated fish lipid oxidation has largely increased in the past 5 years. Although cod light muscle has a low level of intrinsic lipid oxidation catalysts, a prerequisite for a good oxidation model system, we believe it cannot fully mimic the oxidation kinetics taking place in other fish species being more susceptible to lipid oxidation. The aim of this study was to systematically investigate whether washed mince model systems useful in Hb-mediated oxidation studies could be prepared also from herring (Clupea harengus) and salmon (Salmo salar) light muscles. The kinetics of oxidation in the washed models was measured during ice storage (+/-Hb), and the results were related to compositional differences. Minces from cod, herring, and salmon light muscles were washed 3 times with 3 volumes of water and buffer. A 20 microM portion of Hb and 200 ppm streptomycin was then added, followed by adjustment of pH and moisture to 6.3 and 86%, respectively. Samples with or without Hb were then stored on ice, and oxidation was followed as peroxide value (PV), rancid odor, redness (a*) loss and yellowness (b*). Prior to storage, all minces and models were also analyzed for total lipids, fatty acids, alpha-tocopherol, proteins, Hb, Fe, Cu, and Zn. Hb-mediated lipid oxidation appeared within 2 days on ice in all models. Small differences in the oxidation rates ranked the models as herring > cod > salmon. These differences were ascribed to more preformed peroxides and trace elements in the herring model, and more antioxidants in the salmon model. Controls, without Hb, stayed stable in all cases except herring, where a very slight oxidation appeared, especially if the herring raw material had been prefrozen. In conclusion, fattier fish like dark muscle species and salmonoids are useful for making washed mince model systems and would be a better choice than cod if there is an interest in the oxidation kinetics of such species.     

Endothelial cell activation and neovascularization are prominent in dermatomyositis

Background

While vascular and immune abnormalities are common in juvenile and adult dermatomyositis (DM), the molecular changes that contribute to these abnormalities are not clear. Therefore, we investigated pathways that facilitate new blood vessel formation and dendritic cell migration in dermatomyositis.

Methods

Muscle biopsies from subjects with DM (9 children and 6 adults) and non-myositis controls (6 children and 7 adults) were investigated by immunohistochemistry using antibodies that recognize existing (anti-CD146) and newly formed blood vessels (anti-αVβ3) and mature dendritic cells (anti-DC-LAMP). Blood vessel quantification was performed by digitalized image analysis. Additional muscle biopsies from subjects with adult DM and non-myositis controls were used for global gene expression profiling experiments.

Results

A significant increase in neovascularization was found in muscle biopsies of DM patients; neovascularization (αVβ3 positive capillaries and vessels per muscle fiber) was much higher in juvenile than in adult DM patients (control vs juvenile DM: Mean ± SE: 0.06 ± 0.01 vs 0.6 ± 0.05; p < 0.0001 and control vs adult DM: Mean ± SE: 0.60 ± 0.1 vs 0.75 ± 0.1; p = 0.051). Gene expression analysis demonstrated that genes that participate not only in angiogenesis but also in leukocyte trafficking and the complement cascade were highly up regulated in DM muscle in comparison to age matched controls. DC-LAMP positive dendritic cells were highly enriched at perivascular inflammatory sites in juvenile and adult DM patients along with molecules that facilitate dendritic cell transmigration and reverse transmigration (CD142 and CD31).

Conclusion

These results suggest active neovascularization and endothelial cell activation in both juvenile and adult DM. It is likely that close association of monocytes with endothelial cells initiate rapid dendritic cell maturation and an autoimmune response in DM.