Assessment of PCR-based tools for the specific identification of some temperate Meloidogyne species including M. chitwoodi,M. fallax and M. minor

Several conventional PCR tests have been developed for the identification of the European quarantine root-knot nematodes Meloidogyne chitwoodi and M. fallax but data are lacking for the evaluation of their performance in terms of sensitivity, repeatability, reproducibility and specificity against a large range of populations. This study evaluated the performance criteria of three conventional PCR tests recommended by the consensus diagnostic protocol for Meloidogyne chitwoodi and Meloidogyne fallax published by the European and Mediterranean Plant Protection Organization (EPPO): a species-specific PCR (IGS target), a SCAR PCR, and a rDNA ITS PCR-RFLP. Evaluation was carried out with DNA extracts from juveniles, males and females according to EPPO recommendations for test validation. A minimum of 34 populations of target and non target nematode species were tested to check the specificity of these three PCR assays. The three PCR tests were ranked according to their specificity (with regard to cross reaction with other nematodes species or genus) and their sensitivity (detection of a single juvenile or mixed with other species). The species-specific PCR proved to be more sensitive but less specific than the SCAR PCR. The PCR-RFLP enables the identification of several Meloidogyne species but profile analysis can be difficult when several species are present in the mixture. Specific PCR products and RFLP profiles were also observed for M. arenaria and M. enterolobii, and described for M. minor and M. artiellia.     

Prospective echocardiographic and tissue Doppler screening of a large Sphynx cat population: Reference ranges,heart disease prevalence and genetic aspects

Objectives(1) To investigate heart morphology and function using echocardiography and tissue Doppler imaging (TDI), (2) to determine heart disease prevalence and characteristics, and (3) to assess potential genetic features in a population of Sphynx cats presented for cardiovascular screening.AnimalsA total of 147 echocardiographic examinations, including 33 follow-ups, were performed by trained observers on 114 Sphynx cats of different ages (2.62 ± 1.93 years [0.5–10.0]) from 2004 to 2011.MethodsSphynx cats underwent a physical examination, conventional echocardiography, and, if possible, two-dimensional color TDI.ResultsConventional echocardiographic findings included 75/114 normal (65.8%) and 39/114 (34.2%) abnormal examinations with a diagnosis of either congenital heart diseases (n = 16) or hypertrophic cardiomyopathy (HCM, n = 23). In adult healthy cats, a significant body weight effect was observed for several echocardiographic variables, including end-diastolic left ventricular (LV) free wall (P < 0.01), interventricular septum (P < 0.001), and LV diameter (P < 0.001). Mitral valve dysplasia (MVD) was observed as a single or associated defect in 15/16 cats with congenital heart diseases. A significant increase in HCM prevalence (P < 0.001) was observed according to age. The pedigree analysis of a large family (n = 81) suggested an autosomal dominant mode of inheritance with incomplete penetrance for HCM.ConclusionsBody weight should be taken into account when interpreting values of diastolic myocardial wall thicknesses in Sphynx cats. Additionally, HCM and MVD are two relatively common heart diseases in this feline breed. More pedigree data are required to confirm the inheritance pattern of HCM at the breed level.     

Farms, fires, and forestry: Disturbance legacies in the soils of the Northwest Wisconsin (USA) Sand Plain

We studied the long-term effects of disturbance within the Northwest Wisconsin (USA) Sand Plain (NWSP), an ecoregion that is characterized by very sandy soil and an active disturbance history that includes fire, agriculture and industrial forestry, largely clearcut logging of jack pine (Pinus banksiana) and aspen (Populus spp.). Open “barrens” communities on this landscape were formerly maintained by fire, and are a high conservation priority. Hill's Oak (Quercus ellipsoidalis) can also dominate forest canopies, while blueberry (Vaccinium angustifolium), and sweetfern (Comptonia peregrina) are common shrub species. We structured a field sampling design with a spatial-temporal database built from historic airphotos (1938 and 1997) and fire records to examine whether soil organic matter and nutrients vary with disturbance history in the nonforest habitats of the sand plain. We sampled soils along 83 transects, randomly stratified among five sampled classes: (1) nonforest-farming history; (2) nonforest-fire history; (3) nonforest-clearcut only history; (4) evergreen forest of jack pine and red pine (P. resinosa); and (5) deciduous forest of Hill's oak and aspen. Logging of the original forest took place in the late 1800s–early 1900s. The farms were abandoned between 1938 and 1960, and the most recent fire occurred in 1977. Thus, the duration of the agricultural legacy is approximately 45–65 years while observed fire effects have lasted for 26 years.We observed strong agricultural legacies, including high P and low OM, N and Ca. One possible explanation for the N legacy is that it is tied to soil OM accretion which may be driven by plant growth. We detected no difference in mean values for any of the soil properties between soils from nonforested areas within the Five-Mile fire and soils from nonforested areas with a clearcut-only history. We did observe a fire effect in high variance for soil P. This could have resulted from variations in fire severity and ash convection and deposition.Forest soils generally had lower pH than the nonforest soils, and the deciduous forest soils had the lowest pH and also very low Ca. We also observed high within-transect coefficient of variation for Ca in the forest soils.We conclude that agriculture is a qualitatively different disturbance-type than fire or clearcutting, that disturbance legacies tend to be most persistent with geologically stable elements, such as P, and that management and conservation planning within the NWSP would benefit from site-specific agricultural history, as well as attention to Ca.     

Effect of thiamethoxam on cockroach locomotor activity is associated with its metabolite clothianidin

BACKROUND: In the present study, the effect of thiamethoxam and clothianidin on the locomotor activity of American cockroach, Periplaneta americana (L.), was evaluated. Because it has been proposed that thiamethoxam is metabolised to clothianidin, high‐performance liquid chromatography coupled with mass spectrometry was used to evaluate the amount of clothianidin on thiamethoxam‐treated cockroaches. RESULTS: One hour after neonicotinoid treatment, the time spent in the open‐field‐like apparatus significantly increased, suggesting a decrease in locomotor activity. The percentage of cockroaches displaying locomotor activity was significantly reduced 1 h after haemolymph application of 1 nmol g^?1 neonicotinoid, while no significant effect was found after topical and oral administration. However, at 24 and 48 h, all neonicotinoids were able to reduce locomotor activity, depending on their concentrations and the way they were applied. Interestingly, it was found that thiamethoxam was converted to clothianidin 1 h after application, but the amount of clothianidin did not rise proportionately to thiamethoxam, especially after oral administration. CONCLUSION: The data suggest that the effect of thiamethoxam on cockroach locomotor activity is due in part to clothianidin action because (1) thiamethoxam levels remained persistent 48 h after application and (2) the amount of clothianidin in cockroach tissues was consistent with the toxicity of thiamethoxam. Copyright © 2010 Society of Chemical Industry     

Genome-wide association mapping for root cone angle in rice

Background

Plant root systems play a major role in anchoring and in water and nutrient uptake from the soil. The root cone angle is an important parameter of the root system architecture because, combined with root depth, it helps to determine the volume of soil explored by the plant. Two genes, DRO1 and SOR1, and several QTLs for root cone angle have been discovered in the last 5 years.

Results

To find other QTLs linked to root cone angle, a genome-wide association mapping study was conducted on two panels of 162 indica and 169 japonica rice accessions genotyped with two sets of SNP markers (genotyping-by-sequencing set with approximately 16,000 markers and high-density-rice-array set with approximately 300,000 markers). The root cone angle of all accessions was measured using a screen protractor on images taken after 1 month of plant growth in the Rhizoscope phenotyping system. The distribution of the root cone angle in the indica panel was Gaussian, but several accessions of the japonica panel (all the bulus from Indonesia and three temperate japonicas from Nepal or India) appeared as outliers with a very wide root cone angle. The data were submitted to association mapping using a mixed model with control of structure and kinship. A total of 15 QTLs for the indica panel and 40 QTLs for the japonica panel were detected. Genes underlying these QTLs (+/?50 kb from the significant markers) were analyzed. We focused our analysis on auxin-related genes, kinases, and genes involved in root developmental processes and identified 8 particularly interesting genes.

Conclusions

The present study identifies new sources of wide root cone angle in rice, proposes ways to bypass some drawbacks of association mapping to further understand the genetics of the trait and identifies candidate genes deserving further investigation.

     

Encapsulation of Salmon Peptides in Marine Liposomes: Physico-Chemical Properties,Antiradical Activities and Biocompatibility Assays

Salmon byproducts (Salmo salar) generated by the food chain represent a source of long-chain polyunsaturated fatty acids (eicosapentaenoic acid (EPA): 20:5n-3; docosahexaenoic acid (DHA): 22:6n-3) and peptides that can be used as supplements in food for nutraceutical or health applications, such as in the prevention of certain pathologies (e.g., Alzheimer’s and cardiovascular diseases). The extraction of polar lipids naturally rich in PUFAs by enzymatic processes without organic solvent (controlled by pH-Stat method), coupled with the production of 1 kDa salmon peptides by membrane filtration, allowed the formulation of nanocarriers. The physicochemical properties of the nanoliposomes (size ranging from 120 to 140 nm, PDI of 0.27, zeta potential between −32 and −46 mV and encapsulation efficiency) were measured, and the bioactivity of salmon hydrolysate peptides was assessed (antioxidant and antiradical activity: ABTS, ORAC, DPPH; iron metal chelation). Salmon peptides exhibited good angiotensin-conversion-enzyme (ACE) inhibition activity, with an IC₅₀ value of 413.43 ± 13.12 µg/mL. Cytotoxicity, metabolic activity and proliferation experiments demonstrated the harmlessness of the nanostructures in these experimental conditions.