Species-specific DNA markers for identification of two root-knot nematodes of coffee: Meloidogyne arabicida and M. izalcoensis

Seven root-knot nematodes (RKN), including Meloidogyne exigua, M. incognita, M. paranaensis, M. enterolobii, M. arabicida, M. izalcoensis and M. arenaria are major pathogens of coffee crop in the Americas. Species-specific primers for their identification have been developed for five of them and constitute a fast and reliable method of identification. Here we report a PCR-based assay for specific detection of M. arabicida and M. izalcoensis. Random Amplified Polymorphic DNA fragments specific for these two species were converted into sequence characterized amplified region (SCAR) markers. PCR amplification using the SCAR primers produced a specific fragment of 300 bp and 670 bp for M. arabicida and M. izalcoensis, respectively, which were absent in other coffee-associated Meloidogyne spp. tested. SCAR primers also allowed successful amplification of DNA from single second-stage juveniles (J2), males and females. In addition, these primers were able to unambiguously detect the target species in nematode suspensions extracted from soil and roots samples, in different isolates of the same species or when used in multiplex PCR reactions containing mixtures of species. These results demonstrated the effectiveness of these SCAR markers and their multiplex use with those previously developed for M. exigua, M. incognita, M. paranaensis, M. enterolobii and M. arenaria constitute an essential detection tool. This diagnostic kit will contribute for specific J2 identification of the major RKN infecting coffee from field samples in the Americas.     

Ruminal and morphometric parameters of the rumen and intestines of sheep fed with increasing levels of spineless cactus (Nopalea cochenillifera Salm-Dyck)

Tropical Animal Health and Production - The objective of this study was to evaluate the ruminal parameters (pH, N-NH3, and microbial protein) and morphometry of the rumen and intestine of sheep fed...     

Assessment and up-scaling of CO2 exchange by patches of the herbaceous vegetation mosaic in a Portuguese cork oak woodland

Long-term eddy covariance measurements over a montado oak woodland in southern Portugal have documented a vulnerability to predicted decreases in springtime rainfall, since water availability during spring limits annual CO₂ gain, the growth of fodder for animals, and the production of cork by Quercus suber. The current study examined CO₂ exchange of three different herbaceous vegetation components distributed over montado landscapes and within the footprint of long-term landscape eddy covariance monitoring studies. Simultaneous measurements with eddy covariance at two sites and with manually operated chambers at multiple locations revealed that slow drainage of shallow basins, the onset of drying at higher sites and a high release of CO₂ below tree canopies significantly influenced the overall course of montado ecosystem gas exchange during the spring.Hyperbolic light response models were employed to up-scale and compare herbaceous gas exchange with landscape net ecosystem CO₂ flux. The up-scaling demonstrates the importance of the herbaceous understory in determining annual carbon balance of the montado and suggests a relatively small additional CO₂ uptake by the tree canopies and boles, i.e., by the aboveground tree compartment, during springtime. Annual flux totals obtained during the extremely dry year 2005 and a normal precipitation year 2006 for the oak woodland and a nearby grassland were essentially the same, indicating that both ecosystems similarly exploit available resources. Based on comparisons with additional temperate grasslands, we can visualize the montado herbaceous cover as a typical European grassland canopy, but where temperature fluctuations in winter control uptake, and where total production depends on springtime rainfall as it controls phenological events and eventually dieback of the vegetation. On the other hand, tree canopies remain active longer during late spring and early summer, modifying the montado response from that of grassland. Uncertainties in flux estimates via both chamber and eddy covariance methodologies currently prevent a full understanding of vegetation/atmosphere coupling, of the recycling of CO₂ between the understory communities and trees, and of relationships between exchange rates of individual components of the vegetation mosaic and overall carbon and water balances in montado landscapes.     

Horse resistance to natural infestations of Anocentor nitens and Amblyomma cajennense (Acari: Ixodidae).

The objective of this study was to investigate some aspects of horse resistance to natural infestations of Anocentor nitens and Amblyomma cajennense over a 2-year period. Free-living stages of A. nitens were used to evaluate the influence of season of the year on horse resistance. Every 2 weeks, 10 selected engorged females, weighing 150-329 mg, were removed from naturally infested horses and were individually placed into glass tubes closed with a cotton stopper, and maintained in an incubator. The biological parameters evaluated varied according to the season of the year, i.e., reproductive efficiency, maximum larval survival (MLS) and egg-hatching rates were significantly lower in spring/summer than in fall/winter. At 14-day intervals, females of A. nitens and A. cajennense were counted on each side of the body of five mares. A comparison between the animal with the lowest infestation and the animal with the highest infestation showed greater differences in animals infested with A. nitens than those infested with A. cajennense. Ten animals were found to carry 64.9% of the total parasite burden of A. cajennense and 88.6% of the highest infestations of A. nitens females. A seasonal variation on intensity of horse resistance to A. nitens was observed during the study. However, this was not observed for A. cajennense. Cross-resistance was observed in animals with the highest total number of A. cajennense and the highest number of A. cajennense adults (r=0.817), larvae (r=0.756), and A. nitens females (r=0.799). These results led to the conclusion that changes in horse resistance to A. nitens occur seasonally. As far as the total population of A. cajennense and the female stage of A. nitens are concerned, horses develop cross-resistance.     

Nitrogen dynamics during ecosystem development in tropical forest restoration

We considered whether ecological restoration using high diversity of native tree species serves to restore nitrogen dynamics in the Brazilian Atlantic Forest. We measured δ¹⁵N and N content in green foliage and soil; vegetation N:P ratio; and soil N mineralization in a preserved natural forest and restored forests of ages 21 and 52 years. Green foliage δ¹⁵N values, N content, N:P ratio, inorganic N and net mineralization and nitrification rates were all higher, the older the forest. Our findings indicate that the recuperation of N cycling has not been achieved yet in the restored forests even after 52 years, but show that they are following a trajectory of development that is characterized by their N cycling intensity becoming similar to a natural mature forest of the same original forest formation. This study demonstrated that some young restored forests are more limited by N compared to mature natural forests. We document that the recuperation of N cycling in tropical forests can be achieved through ecological restoration actions.     

Can Non-fumigant Nematicides Be an Alternative to Fumigation on Carrot Fields?

ABSTRACT

Root-knot nematodes (RKN; Meloidogyne spp.) cause enormous yield reductions in vegetable crops. While nematicides are the most effective method for controlling RKN in carrot (Dacatus carota), the efficacy of non-fumigant nematicides, compared to fumigants, is still not fully understood. The objective of this study was to evaluate the effect of 1,3-dichloropropene, a fumigant nematicide, and fluensulfone, a non-fumigant nematicide, on RKN control and yield of fresh market and processing carrot. Field experiments were conducted in a commercial farm in Lenox, GA. A randomized complete block design (r = 4) was used in a factorial arrangement of 3 nematicide treatments and 2 carrot cultivars. Nematicide treatments were the application of 1,3-dichloropropene, fluensulfone, and untreated control. Carrot cultivars were Baltimore (fresh marketing) and Belgrado (processing). In general, RKN population was low early season, averaging 0.8 RKN 100 cm^?3 of soil. RKN populations increased with crop development and there were significant differences among nematicide treatments. At harvest, RKN populations were lower for fluensulfone (1.1 RKN 100 cm^?3 of soil) and 1,3-dichloropropene (3.2 RKN 100 cm^?3 of soil) compared to Control (26.4 RKN 100 cm^?3 of soil); however, fluensulfone (91,278 kg ha^?1) had higher yield than 1,3-dichloropropene (64,154 kg ha^?1) and Control (61,908 kg ha^?1). Carrot cultivars were similar for RKN population and yield. Overall, the non-fumigant nematicide was more beneficial for carrot production than the fumigant nematicide.     

Copper and Chromium Electrodialytic Migration in CCA-Treated Timber Waste

Chromated copper arsenate (CCA)-treated timber waste is hazardous for the environment due to the leakage of copper, chromium as well as arsenic to the soil and the groundwater. The study concentrates on electrodialytic removal of copper and chromium and their migration behavior in the wood chips and sawdust matrix in an applied electric field. The method is based on the movement of the charged particles in the electric field, enhanced by incubation with oxalic acid (2.5% w/w) and various placement of the contaminated waste in the electrodialytic cell area to analyze the distribution of chemicals in the proposed matrix. The method was tested in laboratory scale experiments where the CCA contaminated wood chips and clean sawdust were placed in various places of the cell, i.e., adjacent to the cathode and anode compartments and in the mid-section of the cell. The duration of each experiment was 14 days and the current was kept constant at 40 mA. During the electrodialytic treatment negative copper and chromium compounds were mostly found in the anolyte or accumulated in the anion-exchange membrane. The amount of positive compounds found in the catholyte is negligible. The placement of contaminated chips close to the anode and cathode showed that copper and chromium moved towards the positively charged anode. If placed in the mid section of the cell, significant amount of contaminants remained bound to the wood matrix or slightly migrated towards the anode. To predict the formation of copper and chromium species at certain pH and Eh, the computer-modeling program MTDATA to generate Pourbaix diagrams was used.     

Heavy Metal Contamination in Sediments of Lake Nahuel Huapi,Nahuel Huapi National Park,Northern Patagonia,Argentina

Trace metals (Sb, As, Br, Cs, Co, Cr, Ag and Hg), rare earth elements (Ce, Eu, La, Lu, Sm, Tb and Yb), and Sc. were analysed in suspended load and in two sediment cores from Lake Nahuel Huapi, Nahuel Huapi National Park, Patagonia, Argentina, by using Instrumental Neutron Activation Analysis. The core activity profiles of ²¹⁰Pb and ¹³⁷Cs were measured to estimate the age of the sediments. Silver was enriched in the upper layers of both cores, and correlated with the strong growthof the population of Bariloche City in the last 50 yr. Concentrations are higher in the core sampled near Bariloche, andindicate some transport in the water body. Silver concentrationsmeasured in the suspended load were 4.58±0.36 and 3.46±0.40 μg g^-1 in the southern margin of the lake, near San Carlos de Bariloche City, whereas in the northern margin the concentrations were 1.68±0.18 and 1.88±0.23 μg g^-1. Concentration ratio for silver in suspended load and upper core layers were equal in both sampling points. These facts suggest that Ag contents correspond to inputs associated with human activities. Bromine concentrations show a strong increase abovebase line values, similar to silver. In contrast, caesium, chromium, and cobalt concentrations do not differ from base linevalues. The metalloids antimony and arsenic show little variationwith regard to base line concentration. Noticeable, the mercurycontents found in the suspended load.     

A novel vasodilatory peptide from the salivary glands of the sand fly Lutzomyia longipalpis

Salivary gland lysates of the sand fly Lutzomyia longipalpis contain a potent vasodilator that aids the fly to feed on the blood of its vertebrate hosts. Chromatographic analysis, antibody reactivity, and data obtained from bioassays of the salivary erythema-inducing factor indicate striking similarity with human calcitonin gene-related peptide. The erythema-inducing factor is, however, at least one order of magnitude more potent than calcitonin gene-related peptide.