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.     

Global and Phylogenetic Distribution of Quorum Sensing Signals,Acyl Homoserine Lactones,in the Family of Vibrionaceae

Bacterial quorum sensing (QS) and the corresponding signals, acyl homoserine lactones (AHLs), were first described for a luminescent Vibrio species. Since then, detailed knowledge has been gained on the functional level of QS; however, the abundance of AHLs in the family of Vibrionaceae in the environment has remained unclear. Three hundred and one Vibrionaceae strains were collected on a global research cruise and the prevalence and profile of AHL signals in this global collection were determined. AHLs were detected in 32 of the 301 strains using Agrobacterium tumefaciens and Chromobacterium violaceum reporter strains. Ethyl acetate extracts of the cultures were analysed by ultra-high performance liquid chromatography-high resolution mass spectrometry (MS) with automated tandem MS confirmation for AHLs. N-(3-hydroxy-hexanoyl) (OH-C6) and N-(3-hydroxy-decanoyl) (OH-C10) homoserine lactones were the most common AHLs found in 17 and 12 strains, respectively. Several strains produced a diversity of different AHLs, including N-heptanoyl (C7) HL. AHL-producing Vibrionaceae were found in polar, temperate and tropical waters. The AHL profiles correlated with strain phylogeny based on gene sequence homology, however not with geographical location. In conclusion, a wide range of AHL signals are produced by a number of clades in the Vibrionaceae family and these results will allow future investigations of inter- and intra-species interactions within this cosmopolitan family of marine bacteria.     

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.     

The effect of spikelet position on the success frequencies of wheat haploid production using the maize cross system

Barbela is an old Portuguese landrace of wheat that is highly genetically heterogeneous. Different Barbela populations when subjected to aluminium stress show variable levels of tolerance. In order to study the inheritance of this character, doubled haploid (DH) lines were developed. These DH were obtained by intergeneric crosses of 14 different lines of Barbela with maize. During this process the efficiency of the technique was evaluated and suggestions for its improvement were obtained. Several parameters were studied in the crosses: % of crossability, % of embryos per florets pollinated and % of embryos per seed set. The different genotypes of Barbela showed significant variation for the parameters analysed. When the reciprocal crosses were analysed, no differences were found, indicating that cytoplasm differences do not influence the parameters of DH production. However, different spikelet positions (lower, middle and upper) gave highly significant differences in all parameters analysed. Highest success frequencies were obtained for pollinated spikelets in the middle of the spike. This can indicate that concentrating on the middle part of the spike can increase the frequency of DHs obtained using inter generic crosses of wheat with maize. This revised version was published online in August 2006 with corrections to the Cover Date.     

Implications of the number of years assessment on recommendation of common bean cultivars

In the common bean crop in Brazil, the requirement of the value for cultivation and use trials is that these experiments must be conducted over two years in three locations per region. Information in regard to the necessary number of years to ensure precision in cultivar recommendation and the influence of evaluated years in the estimation of the GE interaction are still scarce. Using grain yield of five check varieties assessed over 11 years in three sowing seasons per year, the aims of this study are as follows: to measure the relative contribution of the GE interaction, evaluating the implication of the number of years in the estimates of the GE interaction, and infer how many years are needed to ensure precision in cultivar recommendation. For instances, analysis of variance was carried out involving all environments and also combinations of years. The results showed that the GE interaction was greater than all other cross‐effects involving lines. The use of at least two years allows good coincidence in cultivar recommendation compared to the whole period. Increasing the evaluation time is a good strategy, especially when it is difficult to grow three different sowing seasons.     

The effect of aqueous extract of gross and commercial yerba mate (Ilex paraguariensis) on intra-abdominal and epididymal fat and glucose levels in male Wistar rats

This study analyzed the plasma lipid profile, glucose levels and fat deposits in male rats treated with aqueous extract of gross yerba mate, commercial yerba mate or water. Yerba mate treatment did not change body weight gain and lipid profile. The consumption of gross yerba mate significantly increased blood glucose (6.6 mmol/L) as compared to the water (4.8 mmol/L) and commercial group (5.2 mmol/L) and decreased epididymal and intra-abdominal deposits (10.1 mg/g and 23.7 mg/g of weight) as compared to the water (15.4 mg/g and 36.9 mg/g of weight) and commercial group (12.5 mg/g and 28 mg/g of weight). The results suggest that gross yerba mate reduces fat more efficiently but produces a greater increase in blood glucose when compared to commercial yerba mate and water groups.