Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.     

Ultrastructural Morphology and Nuclear Maturation Rates of Immature Equine Oocytes Vitrified with Different Solutions and Exposure Times

Ultrastructural morphological injuries and maturation rates were investigated in equine oocytes exposed to vitrification solutions (VS) containing synthetic ice blockers (SIBs) during different exposure times. In experiment 1, compact cumulus-oocyte complexes (COCs; n = 30) were randomly allocated to treatments: (1) fresh fixed (control); (2) VS-1 (1.4 M dimethyl sulfoxide [DMSO] + 1.8 M ethylene glycol [EG] + 1% SIB) for 3 minutes of equilibrium time and VS-2 (2.8 M DMSO + 3.6 M EG + 0.6 M sucrose + 1% SIB) for 1 minute (Eq-long); and (3) VS-1 for 1.5 minutes and VS-2 for 30 seconds (Eq-short). In experiment 2, compact (n = 248) and expanded (n = 264) COCs were evenly distributed to the following treatments: (1) immediate maturation in vitro (control); (2) vitrification using the Eq-short protocol as in experiment 1; and (3) vitrification using a stock solution containing 2.8 M formamide, 2.8 M DMSO, 2.7 M EG, 7% polyvinylpyrrolidone, and 1% SIB (Eq-short-mod). More (P < .02) oocytes with normal ultrastructural morphology were seen in fresh control and Eq-short groups than in Eq-long group. Metaphase-II (MII) rates were higher (P < .05) for oocytes with expanded cumulus than compact cumulus in the control group, and higher (P < .05) for oocytes with expanded cumulus than compact cumulus in Eq-short and Eq-short-mod groups. No difference in MII rates was detected among groups within each type of COC. In conclusion, reduction of exposure time to VS better preserved oocyte ultrastructural features, and MII rates were higher for vitrified oocytes with expanded cumulus. This study advances our knowledge on potential alternatives for vitrification of immature equine oocytes.     

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...     

Land Cover Type and Fire in Portugal: Do Fires Burn Land Cover Selectively?

The purpose of this study is to investigate if, or under what conditions, fires select given land cover types for burning. If fires burn unselectively then the land cover composition (the proportional area of various land cover types) of individual fires should approximate the land cover composition available in their neighborhood. In this study we test this hypothesis by performing statistical analyses of a data set consisting of paired vectors with the proportions of land cover types present in burned areas and in their respective surroundings. The statistical methods employed (a permutation technique and the C_max statistic) are commonly used in resource selection studies where data is subject to a unit-sum constraint. The results of the analysis of 506 fires that burned in Portugal in 1991 indicate that fires are selective, with small fires exhibiting stronger land cover preferences than large fires. According to the results of a multiple comparison analysis performed for small fires, there is a marked preference for shrubland followed by other forest cover types, while agriculture is clearly avoided. A similar analysis is performed to test if fire selectivity is related to the ecological region where it occurs. The results obtained in this study contribute to the discussion on the relative importance of fuels as a drivers of fire spread.     

Effects of deficit irrigation strategies on cluster microclimate for improving fruit composition of Moscatel field-grown grapevines

The grapevine plays a very important role in the economic, social and cultural sectors of many regions; however vineyards are often grown in regions under stressful conditions and thus they are vulnerable to climate change. The objective of this research was to investigate the effect of partial root-zone drying (PRD) irrigation on vine water relations, vegetative growth, plant microclimate, berry composition and yield components, compared to conventional deficit irrigation (DI, 50% ETc), full irrigation (FI, 100% of ETc) and non-irrigated vines (NI). The study was undertaken in mature ‘Moscatel’ grapevines (Vitis vinifera L.) grown in Pegões, South of Portugal. Compared to the other irrigated treatments, PRD vines showed a better microclimate at the cluster zone with higher incident photosynthetic photon flux density (PPFD). Within the more open canopies of NI and PRD treatments, berry temperatures were higher than those of denser ones (DI and FI). Compared to the conventional irrigation technique the better microclimate observed in PRD vines was a consequence of a reduction in vine growth, where lower values of leaf layer number, leaf area, canopy wideness, water shoots and shoot weight were observed. In PRD vines we observed a tendency to a development of a deeper root system, while DI and FI showed a more homogeneous root distribution throughout the different soil layers. PRD showed an improvement in berry quality with higher values of flavour precursors, and total phenols concentration without any significant yield reduction compared to DI and FI.     

Dynamics and efficacy of sulfentrazone,flumioxazin, and isoxaflutole herbicides applied on eucalyptus harvest residues

New Forests - Eucalyptus harvests generate residues that remain on the soil and affect the efficacy of preemergent herbicides in the following planting cycle. The objective of this work was to...     

Stem modeling and simulation of conversion of cork oak stems for quality wood products

Recent research is underway to study cork oak (Quercus suber L.) wood potential for production of wood components. A total of 35 stems of young and mature cork oaks were sampled, live sawn into flitches, scanned using VTT’s WoodCIM^®, and the measured data computed by VTT’s PuuPilot software, allowing stem 3D reconstruction. Sawing simulations were run for 0.5 m and 1 m logs and the whole stem. Sawn products were (1) planks, (2) parquet, (3) lamparquet, and (4) external component of multilayer planks. Cork oak stems showed a moderate to small taper (mean 24 mm/m). Curviness varied between straight to significantly crooked stems (mean value 40 mm) Batch yields for the tested products ranged 25–43% and 37–50% for 0.5 m logs of young and mature trees, respectively; for 1 m logs, batch yields ranged 19–41% and 25–54%. When using the whole stem, batch yields were lower, ranging 11–38% and 15–50%. Higher yields were obtained for all log lengths and samples for production of lamparquet, parquet, and multilayer component.     

A case study of <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> fingerprinting for breeding

• Introduction  

Tree genetic improvement programs usually lack, in general, pedigree information. Since molecular markers can be used to estimate the level of genetic similarity between individuals, we genotyped a sample of a Portuguese Eucalyptus globulus breeding population—a reference population of 125 individuals—with 16 microsatellites (SSR).