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.     

Investigations of the pore-size distribution of wood in the dry and wet state by means of mercury intrusion porosimetry

This paper deals with the determination of the pore-size distribution of untreated and thermally modified twin samples of Norway spruce (Picea abies (L.) Karst.) and sycamore maple (Acer pseudoplatanus L.) by means of mercury intrusion porosimetry. The investigations considered oven-dried and at 22 °C and 95 % relative humidity-conditioned samples. A special sample holder and a novel method for evacuation were developed for the measurements. This was necessary both to ensure that samples were positioned at a defined distance and the conditioned samples did not dry in an uncontrolled manner. The results clearly show that the climatic state influences the results. Thermal modification greatly alters the pore-size distribution of spruce. This can be largely attributed to the formation of intercellular spaces in the middle lamella as a result of cell-wall compression due to thermal modification. The alteration of the pore structure of maple due to thermal modification is not as pronounced as that of spruce.     

Genetic structure of Orbicella faveolata population reveals high connectivity among a marine protected area and Varadero Reef in the Colombian Caribbean

1. The protection of ecosystems with high diversity, such as coral reefs, is not an approach that guarantees their conservation. Thus, maintaining connectivity among coral populations over the long term is a strategy that should be adopted in order to protect diversity and ecological processes. Although coral reefs in Colombia are highly diverse, the population genetics baseline data of keystone species such as Orbicella faveolata are limited.
2. To provide current information about the connectivity between populations of O. faveolata in the Colombian Caribbean, the genetic diversity and distribution of O. faveolata was evaluated with analysis of molecular variance (AMOVA), principal components, and migration estimations using 113 genets distributed in six populations. Also, a genetic structure analysis that included the available data for the Caribbean population was conducted, seeking to understand how the Colombian populations relate to the broader region.
3. According to the global fixation index (F_ST) for the Colombian Caribbean, there was no genetic structure (F_ST = 0.002). Discriminant analysis of principal components (DAPC) showed that Corales del Rosario Archipelago (CR), Este de Isla Fuerte (EIF), San Bernardo Archipelago (SB), and Varadero Reef (VR) were grouped with Oeste de Isla Fuerte (OIF), with Bushnell (BS) representing the most divergent cluster. Migration analysis showed relatively high migration from VR to CR and SB, highlighting the importance of VR as a genetic reservoir for the region. Structure analysis showed that the Colombian population presented a specific genetic identity (F_ST = 0.254), suggesting that the Colombian Caribbean population could be a peripheral population that contributes significantly to genetic variation and is connected through a complex connectivity process.
4. In conclusion, the estimated genetic connectivity reflects the influence of sea surface dynamics over the interpopulation exchange dynamics and the role of protected and nonprotected coral reef in the Colombian Caribbean. Thus, the Colombian Caribbean population of O. faveolata could be relevant to the conservation of genetic diversity on a larger scale.

     

Assessment of the performance of Pacific oyster (Crassostrea gigas) larvae fed with fresh and preserved Pavlova lutheri concentrates

The Prymnesiophycea Pavlova lutheriwas produced using a traditional technique (batch mode in carboys) and by an innovative technique (semi-continuous mode in alveolarphotobioreactor). Two experimental feedingtrials were carried out with Pacific oyster(Crassostrea gigas) larvae with freshand preserved (4 °C and 1 °C)P. lutheri concentrates used inmonospecific and bispecific diets, incombination with fresh Chaetoceroscalcitrans forma pumilum. Nodifferences in terms of gross composition andlarval performance were noticed between thefresh algae biomass produced by the twotechniques. The low temperatures allowedpreservation of the nutritionalcharacteristics of the P. lutheri freshculture for the time period considered (27days). The P. lutheri concentratespreserved at 4 °C or 1 °C didnot differ substantially in terms ofnutritional value. Larvae fed a monospecificdiet of P. lutheri, both fresh andpreserved, exhibited a modest growth rate (<2.1 µm day^–1), in some cases notsignificantly different from that observed inthe unfed larvae (the control), and a lowsurvival rate. In contrast, when Chaetoceros calcitrans forma pumilumwas used as a monospecific diet, good growthperformance (>4.5 µm day^–1) and ahigh survival rate (>86%) were observed inlarvae. A substitution of 50% (trial 1) or80% (trial 2) of fresh C. calcitransforma pumilum with the preserved P. lutheri concentrates did not adverselyaffect growth rate or survival of C.gigas larvae.     

Fate of C- and N-labelled rhizodeposition of Lolium perenne as function of the distance to the root surface

A greenhouse rhizobox experiment was carried out to quantify the incorporation of ¹³C- and ¹⁵N-labelled rhizodeposits into different soil pools, especially into the rhizosphere microbial biomass, with increasing distances to the root surface of Lolium perenne. Five layers were analysed over 0-4.2 mm distance to an artificial root surface. C and N derived from rhizodeposition were 4.2% of total C and 2.8% of total N in soil at 0-1.0 mm distance and decreased rapidly with increasing distance. Microbial biomass C and N increased significantly towards the roots. At 0-1.0 mm distance microbial biomass C and N accounted for 66% and 29% of C and N derived from rhizodeposition, respectively. These percentages declined with increasing distance to the roots, but were still traceable up to 4.2 mm distance. Only small amounts of root released C and N were found in the 0.05 M K₂SO₄-extractable fraction. Extractable C and N derived from rhizodeposition varied around means of 4% of total C and N derived from rhizodeposition and increased only marginally with increasing distance to the roots. C derived from rhizodeposition in the non-extractable soil organic matter increased from 65 to 89% of total C derived from rhizodeposition at 0-3.4 mm distance. Conversely, microbial biomass C derived from rhizodeposition decreased from 33 to 4%. N derived from rhizodeposition in the non-extractable soil organic matter increased from 61 to 79% of total N derived from rhizodeposition at 0-2.6 mm distance, followed by a decline to roughly 55% in the two outer layers. Microbial biomass N decreased from 37 to 16% at 0-2.6 mm distance, followed by an increase to roughly 41% in the two outer layers. The C/N ratio of total C and N derived from rhizodeposition as well as that of extractable C and N derived from rhizodeposition increased with increasing distance to the roots to values above 30. In contrast, the C/N ratio of incorporated rhizodeposition C and N into the microbial biomass decreased to values less than 5 at 2.6-4.2 mm distance. The data indicate differential microbial response to C and N derived from rhizodeposition at a high spatial resolution from the root surface. The turnover of C and N derived from rhizodeposition in the rhizosphere as a function of the distance to the root surface is discussed.     

Copper Biosorption by Biomass of Marine Alga: Study of Equilibrium and Kinetics in Batch System and Adsorption/Desorption Cycles in Fixed Bed Column

Copper biosorption onto chemically modified biomass of marine alga Sargassum filipendula was investigated in a batch reactor and a fixed bed column. Experiments were carried out in the batch reactor to obtain kinetic and equilibrium data and to assess the copper desorption efficiency of different eluent solutions. The pseudo-first-order, pseudo-second-order, and Langmuir kinetic models were used to correlate kinetic data. The experimental data fitted well to the pseudo first order and Langmuir kinetic models. Langmuir and Freundlich models were applied to describe the equilibrium data obtained at a fixed temperature of 30°C and at pH values of 3.0, 4.0, 5.0, and 6.0. The maximum capacities of copper biosorption onto the algal biomass were 1.43, 1.59, 2.40, and 2.36 mequiv./g at pH 3.0, 4.0, 5.0, and 6.0, respectively. The efficiencies of two eluent solutions (calcium chloride and hydrochloric acid) for copper removal from the biomass were evaluated at different concentrations (0.1, 0.2, 0.5, and 1.0 mol/L). The efficiencies of the calcium chloride solutions varied from 1% to 14%, while efficiencies varying from 95% to 99% were obtained when hydrochloric acid solutions were applied. Three adsorption/desorption cycles were carried out in a fixed bed column using 0.1 mol/L hydrochloric acid as eluent solution. The results showed that an increase in the number of cycles led to a reduction in the adsorption capacity of the alga. The desorbed copper fraction presented no significant variation, remaining around 63% in the three adsorption/desorption cycles.     

Effects of short-term light to heavy exercise on gastric ulcer development in horses and efficacy of omeprazole paste in preventing gastric ulceration

OBJECTIVE: To determine the effects of 8 days of light to heavy exercise on gastric ulcer development in horses and determine the efficacy of omeprazole paste in preventing gastric ulceration. DESIGN: Randomized, controlled, multicenter clinical trial. ANIMALS: 102 horses with normal-appearing gastric mucosa on endoscopic examination that were in light to heavy training. PROCEDURES: Horses at 4 trial locations were allocated into replicates and sham dosed orally (empty syringe) or treated with a paste formulation of omeprazole (1 mg/kg [0.45 mg/ lb], PO) once daily for 8 days. Training regimens varied among locations and included early training for western performance events; walking, trotting, and cantering in a mechanical exerciser; and race training (2 locations). Prevalences of gastric ulceration at the completion of the 8-day treatment period were compared between groups. RESULTS: At the end of the 8-day treatment period, the proportion of omeprazole-treated horses free from gastric ulceration (88%) was significantly higher than the proportion of sham-dosed horses free from gastric ulceration (27%). CONCLUSIONS AND CLINICAL RELEVANCE: Results showed that horses in light to heavy training for as short as 8 days were at risk of developing gastric ulcers and that administration of omeprazole paste decreased the incidence of gastric ulcers.     

Host/parasite relationship in the in vitro infection of rat gliocytes by Neospora caninum: evaluation of cell respiration

Neospora caninum is a protozoon that causes abortion in cattle and neuromuscular lesions in dogs, with the formation of cysts mainly in the central nervous system. Since N. caninum is an intracellular parasite with tropism for the cells of nervous system, this study evaluated the respiratory metabolism of glial cells infected by this* parasite. Glial cultures obtained from the cerebral cortex of newborn rats were kept in DMEM enriched with 10% fetal bovine serum, 1 mM pyruvic acid and 2 mM of L-glutamine. They were infected at a ratio of approximately 1:1 (cell/parasite). Oxygen consumption was evaluated by polarography in the non infected and N. caninum infected groups, 24 and 72 h following infection. Glial cell respiration after 24 and 72 h was 307.2 +/- 34.7 and 308.9 +/- 64.1 microL of oxygen per mug of total protein per minute, and 566.2 +/- 54.6 and 579 +/- 117.5 microL O2/microg of total protein/minute in the control and infected groups, respectively. These results show that N. caninum does not interfere with glial respiration in vitro.