Evidence of Bartonella sp. in questing adult and nymphal Ixodes ricinus ticks from France and co-infection with Borrelia burgdorferi sensu lato and Babesia sp

Ticks are known vectors for a wide range of pathogenic microorganisms. Their role in the transmission of some others is so far only suspected. Ticks can transmit multiple pathogens, however, little is known about the co-existence of these pathogens within questing ticks. We looked for the presence of DNA from three micro-organisms, Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. which are known or suspected tick-borne pathogens, using a cohort of 92 questing Ixodes ricinus ticks collected from pastures in northern France. DNA was extracted from each individual tick and the presence of the three pathogens was investigated using Polymerase Chain Reaction (PCR) amplification. Nine among 92 samples (9.8%) demonstrated PCR products using Bartonella specific primers, 3 among 92 (3.3%) using Borrelia burgdorferi sensu lato specific primers and 19 among 92 (20.6%) using Babesia specific primers. Seven among 92 samples (7.6%) were PCR positive for at least two of the pathogens and one sample was positive for all three. Adult ticks (12/18; 67%) showed significantly higher infection rates compared to nymphs (11/74; 15%) for all three pathogens (P < 0.001). This study is the demonstration of the simultaneous presence of Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. in questing Ixodes ricinus ticks.     

Plasma N-terminal pro-B-type natriuretic peptide concentration helps to predict survival in dogs with symptomatic degenerative mitral valve disease regardless of and in combination with the initial clinical status at admission

Objectives

Although mitral valve disease (MVD) is the most common canine heart disease, readily available prognostic markers of the disease are still lacking. The aim of this study was to evaluate the comparative ability of N-terminal pro-B-type natriuretic peptide plasma concentration (NT-proBNP) and various echocardiographic variables to predict outcome in dogs with MVD.

Animals, materials and methods

Seventy-four dogs with ISACHC classes 2 and 3 MVD (Groups A and B, respectively) were prospectively recruited. NT-proBNP and several echo-Doppler variables at inclusion were compared as predictors of outcome at 6 months in 54/74 dogs.

Results

NT-proBNP was significantly higher in Group B than in Group A (P < 0.0001), and was the only tested variable significantly different between survivor and non-survivor dogs in both groups (P < 0.05). In the whole canine population, a threshold of 1500 pmol/L could discriminate survivor from non-survivor dogs with a sensitivity and specificity of 80% and 73%, respectively. When combining ISACHC class with NT-proBNP levels, a cut-off of 1265 pmol/L was predictive of survival in Group A, whereas the cut-off was 2700 pmol/L for Group B.

Conclusions

NT-proBNP is correlated with MVD severity and could be used in combination with clinical status to predict cardiac outcome.     

Within-day and between-day variability of transthoracic anatomic M-mode echocardiography in the awake bottlenose dolphin (Tursiops truncatus)

The use of transthoracic echocardiography in dolphins has been limited so far owing to technical and anatomical specificities. Anatomic M-mode (AMM) is a postprocessing echocardiographic technique generating M-mode studies from two-dimensional (2D) cineloops independently of the ultrasound beam orientation. The aim of the present study was to determine the within-day (repeatability) and between-day (reproducibility) variability of AMM echocardiography in awake healthy bottlenose dolphins (BN, Tursiops truncatus). Four adult BN trained to lie in left recumbency at the water surface were involved in the protocol. A total of 96 echocardiographic examinations were performed on 4 different days by a trained observer examining each BN 6 times per day. Video clips of 2D left parasternal long-axis views showing the left ventricle (LV) ventrally and the aortic root dorsally were recorded at each examination and analyzed for AMM measurements in a random order. A general linear model was used to determine the within-day and between-day coefficients of variation (CV). All examinations were interpretable allowing calculation of 10 AMM variables (i.e., end-diastolic and end-systolic ventral and dorsal LV myocardial wall thicknesses as well as LV and aortic diameters, mean aortic diameter, and LV shortening fraction). Most within- and between-day CV values (18/20) were <15%, the lowest being observed for the end-diastolic LV diameter (1.6%). In conclusion, AMM provides a simple non-invasive evaluation of heart morphology and function in the awake BN with good repeatability and reproducibility of the measurements. Further studies are required to determine the corresponding reference intervals.     

Transient co-expression with three <Emphasis Type="Italic">O</Emphasis>-glycosylation enzymes allows production of GalNAc-<Emphasis Type="Italic">O</Emphasis>-glycosylated Granulocyte-Colony Stimulating Factor in <Emphasis Type="Italic">N. benthamiana</Emphasis>

Background

Expression of economically relevant proteins in alternative expression platforms, especially plant expression platforms, has gained significant interest in recent years. A special interest in working with plants as bioreactors for the production of pharmaceutical proteins is related to low production costs, product safety and quality. Among the different properties that plants can also offer for the production of recombinant proteins, protein glycosylation is crucial since it may have an impact on pharmaceutical functionality and/or stability.

Results

The pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor was transiently expressed in Nicotiana benthamiana plants and subjected to mammalian-specific mucin-type O-glycosylation by co-expressing the pharmaceutical protein together with the glycosylation machinery responsible for such post-translational modification.

Conclusions

The pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor can be expressed in N. benthamiana plants via agroinfiltration with its native mammalian-specific mucin-type O-glycosylation.

     

In vitro metabolism of progesterone, androgens and estrogens by rainbow trout embryos

The purpose of the study was to determine the steroid metabolic pathways used by rainbow trout (Oncorhynchus mykiss) embryos. Whole embryo preparations at 42, 53 and 65 days post fertilization (dpf) were incubated, in vitro, with the tritium-labelled steroids, progesterone (P₄), testosterone (T), androstenedione (A4), 17β-estradiol (E₂), and estrone (E₁), and the metabolites formed were separated and identified by reverse phase high performance liquid chromatography (HPLC). The degree of metabolism for each of the substrates was dependent on the age of the embryos, and was always higher in older embryos. P₄ yielded three metabolites, one of them identified by gas chromatography-mass spectroscopy as 3β,7α-dihydroxy-5α-pregnan-20-one. Several unknown metabolites of a similar profile on HPLC were produced for T and A₄, in addition to A₄ and T as metabolites of T and A ₄ metabolism, respectively. Likewise, E₂ and E₁ yielded each other and an unknown polar metabolite. These observations argue for the presence of 5α-reductase, 7α-hydroxylase and 17β-hydroxysteroid dehydrogenase activity in embryo tissues which, by their conversion of biologically active yolk steroids of maternal origin, may provide a protective environment for the embryo during early development.     

Combining permanent aerobiological networks and molecular analyses for large-scale surveillance of forest fungal pathogens: A proof-of-concept

Forest disease management relies principally on a preventive approach in which epidemiological surveillance plays a crucial role. However, efficient and cost-effective surveillance methods are not currently available for large spatial scales. Nevertheless, aerobiological networks have been set up for several decades in many countries to monitor pollen dispersal and provide real-time assessments of allergenic risk. Here, we suggest that the same approach could be used for the surveillance of forest pathogens. Using molecular methods, we analysed samples from 12 sites of the French aerobiological network, at different dates. Both metabarcoding by high-throughput sequencing (using two markers and two different bioinformatics approaches) and real-time PCR targeting eight important forest pathogens were conducted. To validate the approach, temporal and spatial trends of spore detection were compared with field disease data. The metabarcoding approach demonstrated that many fungal plant pathogens could be found in aerobiological samples. Moreover, five of the eight targeted forest pathogens were detected by real-time PCR, with temporal and spatial trends of spore capture consistent with field data. In particular, Hymenoscyphus fraxineus was detected at high frequency in aerobiological samples in the areas where ash dieback has been present for the longest period of time, and at lower frequency in areas with more recent invasion. Spore detection of seasonal pathogens showed a temporal pattern similar to that of disease reports. Overall, our study provides a proof of concept that permanent aerobiological networks combined with molecular methods may provide a useful tool for large-scale surveillance of forest pathogens.     

Estimating forest soil bulk density using boosted regression modelling

Soil bulk density (ρ) is an important physical property, but its measurement is frequently lacking in soil surveys due to the time‐consuming nature of making the measurement. As a result pedotransfer functions (PTFs) have been developed to predict ρ from other more easily available soil properties. These functions are generally derived from regression methods that aim to fit a single model. In this study, we use a technique called Generalized Boosted Regression Modelling (GBM; Ridgeway, 2006 ) which combines two algorithms: regression trees and boosting. We built two models and compared their predictive performance with published PTFs. All the functions were fitted based on the French forest soil dataset for the European demonstration Biosoil project. The two GBM models were Model G3 which involved the three most frequent quantitative predictors used to estimate soil bulk density (organic carbon, clay and silt), and Model G10, which included ten qualitative and quantitative input variables such as parent material or tree species. Based on the full dataset, Models G3 and G10 gave R² values of 0.45 and 0.86, respectively. Model G3 did not significantly outperform the best published model. Even when fitted from an external dataset, it explained only 29% of the variation of ρ with a root mean square error of 0.244 g/cm³. In contrast, the more complex Model G10 outperformed the other models during external validation, with a R² of 0.67 and a predictive deviation of ±0.168 g/cm³. The variation in forest soil bulk densities was mainly explained by five input variables: organic carbon content, tree species, the coarse fragment content, parent material and sampling depth.     

Three-year comparison of the content of antioxidant microconstituents and several quality characteristics in organic and conventionally managed tomatoes and bell peppers

Understanding how the environment and production and cultivation practices influence the composition and quality of food crops is fundamental to the production of high-quality nutritious foods. In this 3-year study, total phenolics, percent soluble solids, ascorbic acid, and the flavonoid aglycones quercetin, kaempferol, and luteolin were measured in two varieties of tomato (Lycopersicon esculentum L. cv. Ropreco and Burbank) and two varieties of bell peppers (Capsicum annuum L. cv. California Wonder and Excalibur) grown by certified organic and conventional practices in a model system. Significantly higher levels of percent soluble solids (17%), quercetin (30%), kaempferol (17%), and ascorbic acid (26%) were found in Burbank tomatoes (fresh weight basis; FWB), whereas only levels of percent soluble solids (10%) and kaempferol (20%) were significantly higher in organic Ropreco tomatoes (FWB). Year-to-year variability was significant, and high values from 2003 influenced the 3-year average value of quercetin reported for organic Burbank tomatoes. Burbank tomatoes generally had higher levels of quercetin, kaempferol, total phenolics, and ascorbic acid as compared to Ropreco tomatoes. Bell peppers were influenced less by environment and did not display cropping system differences.     

Rapid discrimination of scented rice by solid-phase microextraction, mass spectrometry, and multivariate analysis used as a mass sensor

This study describes a new and suitable method for the rapid evaluation of rice (Oryza sativa, L.) aroma by analysis of the volatile fraction using solid-phase microextraction coupled with mass spectrometry (SPME/MS). The abundance list of unresolved mass fragments of the SPME extracted volatile fraction formed the "fingerprint" of a rice sample. Fingerprints of 61 rice samples were recorded in duplicate. Pollutants originating from the extraction system induce fingerprint background that could be lowered by careful cleaning of vials and caps and by exclusion of specific mass fragments. A good discrimination between scented and nonscented rice samples was obtained using the SIMCA procedure. Most of the discriminating mass fragments could be directly or indirectly assigned to potential aromatic molecules present in rice.