Localization of CD9 Molecule on Bull Spermatozoa: Its Involvement in the Sperm–Egg Interaction

Tetraspanin CD9 is one of the egg membrane proteins known to be essential in fertilization process. The presence and localization of CD9 molecule in spermatozoa and its possible function in reproduction are still unclear. In our study, we describe the localization of CD9 on bull spermatozoa. In the immunofluorescence assay, the positive signal has been observed in the high proportion of sperm cells as a fine grains either on the apical part or through the entire anterior region of sperm head. CD9 recognized by monoclonal antibody IVA‐50 was detected on freshly ejaculated (83.4 ± 3.7%) and frozen‐thawed (84.3 ± 2.3%) sperm. The same reaction pattern was observed on sperm capacitated for 1 h, 2 h, 3 h and 4 h (83.6 ± 2.0%; 84.0 ± 1.5%; 85.7 ± 0.8%; 77.5 ± 10.8%). The presence of CD9 exclusively on plasma membrane of the bovine sperm has been detected by Western blot analysis of the protein fractions after the discontinuous sucrose gradient fractionation of the bull sperm. Moreover, probable role of the sperm CD9 molecule in fertilization process of cattle has been suggested as sperm treatment with anti‐CD9 antibody significantly reduced (by 25%, p ≤ 0.001) the number of fertilized oocytes compared to control group in fertilization assay in vitro.     

Population pharmacokinetics of methadone hydrochloride after a single intramuscular administration in adult Japanese sika deer (Cervus nippon nippon)

ObjectiveTo assess the population pharmacokinetics of methadone in deer.Study designProspective non-randomized experimental trial.AnimalsTwelve healthy adult sika deer (nine males and three females).MethodsDeer received intramuscular administration of racemic methadone hydrochloride at 0.5 mg kg⁻¹ or 1 mg kg⁻¹. Plasma methadone and its metabolite 2-Ethylidene-1,5-Dimethyl-3,3-Diphenyl-Pyrolidine (EDDP) concentrations were determined by validated liquid chromatography coupled to tandem mass spectrometry methods, at times 0, 30 minutes, 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours. Population pharmacokinetics analysis was undertaken using a non-linear mixed effects modelling (NONMEM).ResultsA two-compartment linear disposition model best described observed time-concentration profiles of methadone and EDDP. Population parameter estimates of methadone were elimination clearance (17.3 L hour⁻¹), metabolic clearance (34.6 L hour⁻¹), volume of distribution of compartment 1 (216.0 L) and volume of distribution of compartment 2 (384.0 L). Population parameter estimates of EDDP were elimination clearance (121.0 L hour⁻¹), volume of distribution of compartment 3 (1.08 L) and volume of distribution of compartment 4 (499.5 L). The total clearance and total volume of distribution of methadone and EDDP were 51.9 L hour⁻¹, 121.0 L hour ⁻¹, 600.0 L and 500.6 L, respectively. The methadone terminal elimination half-life was 8.19 hours. No adverse effects were observed after methadone administration.Conclusions and Clinical relevanceFollowing intramuscular injection, methadone was characterized by a large total volume of distribution, high systemic clearance and intermediate terminal half-life in sika deer.     

The potential environmental impact of pesticides removed from sprayers during cleaning

BACKGROUND: There is concern that pesticide residues on the external surfaces of sprayers could have an adverse impact on the environment if they are washed off, yet there is a need to remove these residues for health reasons. The aim of this study was to quantify pesticide residues contained in washings from cleaning discrete parts of a sprayer and to assess their likely environmental impact. RESULTS: The boom/rear of the sprayer and the spray tank accounted for 80% of the total pesticide load in the washings. Predicted environmental pesticide concentrations from sprayer washings were lower than predictions from the FOCUS surface water model for pesticides used under normal agricultural conditions, although for tebuconazole this difference was smaller than for the other compounds investigated. The field area over which the residues may need to be uniformly deposited to avoid overdosing during infield cleaning was typically less than 0.5 m(2), with a maximum value of 4 m(2). CONCLUSIONS: It is unlikely that infield cleaning will lead to overdosing. External residues are not insignificant, so any adverse impact on the environment must be mitigated. Appropriate measures include cleaning in the field away from surface waters and other sensitive areas, and cleaning machines over bunded areas or similar.     

Functional investigation of a QTL affecting resistance to Haemonchus contortus in sheep

This study reports a functional characterization of a limited segment (QTL) of sheep chromosome 12 associated with resistance to the abomasal nematode Haemonchus contortus. The first objective was to validate the identified QTL through the comparison of genetically susceptible (N) and resistant (R) sheep produced from Martinik × Romane back-cross sheep. The R and N genotype groups were then experimentally infected with 10 000 H. contortus larvae and measured for FEC (every three days from 18 to 30 days post-challenge), haematocrit, worm burden and fertility. Significant differences in FEC and haematocrit drop were found between R and N sheep. In addition, the female worms recovered from R sheep were less fecund. The second step of the characterization was to investigate functional mechanisms associated with the QTL, thanks to a gene expression analysis performed on the abomasal mucosa and the abomasal lymph node. The gene expression level of a candidate gene lying within the QTL region (PAPP-A2) was measured. In addition, putative interactions between the chromosome segment under study and the top ten differentially expressed genes between resistant MBB and susceptible RMN sheep highlighted in a previous microarray experiment were investigated. We found an induction of Th-2 related cytokine genes expression in the abomasal mucosa of R sheep. Down-regulation of the PAPP-A2 gene expression was observed between naïve and challenged sheep although no differential expression was recorded between challenged R and N sheep. The genotyping of this limited region should contribute to the ability to predict the intrinsic resistance level of sheep.     

Low soil temperatures increase carbon reserves in Picea mariana and Pinus contorta

? Context

Soil temperature can limit tree growth and function, but it is often unaddressed in understanding the successional status of trees.

? Aims

We tested how soil temperature affected carbon allocation strategies of two dominant co-occurring boreal conifer species, Pinus contorta and Picea mariana.

? Methods

We measured nonstructural carbon (NSC) concentrations, biomass, and photosynthesis of dormant and actively growing 2-year-old seedlings in response to three soil temperatures (5, 10, and 20 °C) under a common ambient air temperature.

? Results

For both species, variation in carbon reserves with soil temperature was more pronounced following seedling growth than during dormancy. For both species and all organ types (roots, needles, and stems), NSC concentrations were highest when seedlings were grown at 5 than 20 °C. Mass adjusted for NSC content was negatively correlated with NSC concentration for all organ types of both species. Soil temperature had a marginally significant effect on photosynthesis of pine; seedlings grown at 10 or 20 °C acquired more carbon than seedlings grown at 5 °C. Spruce seedlings photosynthesized more when grown at 20 °C than at 5 or 10 °C.

? Conclusion

Interspecific differences in allocation of carbon may underlie the responses of P. mariana and P. contorta to cold soils and consequently their successional status.     

Reporting and advising on the effects of fishing

Scientists hold different views about environmental management. These views may drive their interest in the subject and help them to address a wide range of research issues, but they can also affect the ways in which research results are interpreted and reported. Studies that mix science and perspective can compromise public and scientific understanding of fishing effects, as perceived differences in evidence may actually reflect differences in interpretation. To improve the rigour of ‘fishing effects’ science, it would help if the benchmarks used to assess whether fishing effects ‘matter’ were always made explicit. These benchmarks might be the objectives set by the management authorities and/or a series of alternate objectives proposed and stated by the scientist. To demonstrate how the reported significance of fishing effects can depend on objectives, I use a simple model to predict the response of fish populations and communities to fishing. Fishing effects that would be reported as negative in relation to preservation or biodiversity objectives, such as declines in size, abundance and trophic level, occur at lower fishing intensities than those associated with meeting sustainability objectives for target species. When fishing pressure is so high that both conservation and fisheries objectives are not being met, the initial management actions to meet a range of objectives are likely to be compatible (e.g. reduce capacity, support alternate livelihoods).