Sperm chromatin condensation as an in vivo fertility biomarker in bulls:a flow cytometry approach

Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the years.Thus,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive efficiency.In humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male infertility.The objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in bulls.The study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein males.Reproductive outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow cytometry.Sperm quality parameters(morphology,viability,total and progressive motility)were also assessed.Pearson correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo fertility.Sperm morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile individuals.Alternatively,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in bulls.Specifically,the percentage of viable sperm with poor chromatin condensation showed better accuracy and precision to predict in vivo fertility,with an AUC of 0.78(P=0.02).Conclusions:Chromatin condensation evaluated through CMA3/Yo-Pro-1 and flow cytometry is defined here as a more powerful tool than conventional sperm parameters to predict bull in vivo fertility,with a potential ability to maximising the efficiency of dairy breeding industry.     

Experimental realization of a magnetic cloak

Invisibility to electromagnetic fields has become an exciting theoretical possibility. However, the experimental realization of electromagnetic cloaks has only been achieved starting from simplified approaches (for instance, based on ray approximation, canceling only some terms of the scattering fields, or hiding a bulge in a plane instead of an object in free space). Here, we demonstrate, directly from Maxwell equations, that a specially designed cylindrical superconductor-ferromagnetic bilayer can exactly cloak uniform static magnetic fields, and we experimentally confirmed this effect in an actual setup.     

Conservation of species interaction networks

Recent work has shown that antagonist (e.g. predator-prey food web) and mutualist (e.g. pollinator-plant) network structure can be altered by global environmental change drivers, and that these alterations may have important ecosystem-level consequences. This has prompted calls for the conservation of network structure, but precisely which attributes of webs should be conserved remains unclear. Further, the extent to which network metrics characterise the spatiotemporally-variable dynamic structure of interacting communities is unknown. Here, we summarise the attributes of web structure that are predicted to confer stability or increased function to a system, as these may be of greatest interest to conservation biologists. However, empirical evaluation of these effects is lacking in most cases, and we discuss whether stability is even desirable in all contexts. The incorporation of web attributes into conservation monitoring requires that changes in these attributes can be recorded (sampled) with relative ease. We contrast the sensitivity of metrics to sampling effort, and highlight those (such as nestedness and connectance) that could easily be incorporated into conservation monitoring. Despite our growing understanding of the characteristics of food webs that confer stability and function, numerous practical challenges need to be overcome before the goal of conserving species interaction networks can be achieved.     

Comparison between meloxicam and transdermally administered fentanyl for treatment of postoperative pain in dogs undergoing osteotomy of the tibia and fibula and placement of a uniplanar external distraction device

OBJECTIVE: To compare the efficacy of meloxicam administered perioperatively with transdermal administration of fentanyl via a patch placed preoperatively in dogs undergoing orthopedic surgery. DESIGN: Prospective study. ANIMALS: 16 dogs. PROCEDURE: Unilateral or bilateral osteotomy of the tibia and fibula was surgically performed, and a uniplanar external distraction device was placed in each limb. Postoperative pain and lameness were assessed 24, 48, and 72 hours after administration of the first of 3 doses of meloxicam (0.2 mg/kg [0.09 mg/lb], IV, given preoperatively, followed by 0.1 mg/kg [0.045 mg/lb], IV, after 24 hours, and 0.1 mg/kg, PO, after 48 hours) or preoperative placement of a transdermal fentanyl patch (50 microg/h) left in place for 72 hours. RESULTS: No significant differences in total pain scores were detected between groups. Mean +/- SD lameness scores assessed at 24 and 72 hours were lower in dogs in the meloxicam group than dogs in the fentanyl group. Lameness scores decreased with time in a similar manner in both treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: Perioperative administration of meloxicam or preoperative placement of a transdermal fentanyl patch provided effective and similar postoperative analgesia in dogs undergoing orthopedic surgery. However, because of its anti-inflammatory effects, treatment with meloxicam reduced the degree of lameness and resulted in rapid functional recovery of the limb.     

Development of a genetic manipulation system for Haemophilus parasuis

Haemophilus parasuis is a member of the family Pasteurellaceae and an important respiratory-tract pathogen of swine, which is the etiological agent of Glasser's disease. Because no genetic manipulation system is available for H. parasuis so far, in vivo studies about the role of its genes involved in virulence are unfeasible. Here we demonstrate that H. parasuis has a cyclic AMP (cAMP)-dependent natural transformation system that enables the uptake of DNA in which the ACCGAACTC sequence signal must be present. After improving DNA transformation parameters, such as cAMP and DNA concentration and exposition time of the exogenous DNA, a knockout mutant of H. parasuis defective in the thy gene, encoding the thymidylate synthase enzyme, has been constructed. Data presented in this work open the possibility for the functional analysis of genes involved in the infectious process of this animal pathogen.     

Are protected areas preserving ecosystem services and biodiversity? Insights from Mediterranean forests and shrublands

Landscape Ecology - Protected areas (PAs) are essential for biodiversity conservation and the provision of ecosystem services (ES), representing 15% of the earth’s surface and targeted to...     

Analysis of Aggressiveness of Erwinia amylovora Using Disease-Dose and Time Relationships

ABSTRACT The aggressiveness of an extensive collection of strains of Erwinia amylovora was analyzed using immature fruit and detached pear flower assays under controlled environmental conditions. The analysis was performed by means of a quantitative approach based on fitting data to mathematical models that relate infection incidence to pathogen dose and time. Probit and hyperbolic saturation models were used for disease-dose relationships and provided information on the median effective dose (ED(50)). Values of ED(50) ranged from 10(3) to 10(6) CFU/ml (10 to 10(4) CFU per site of inoculation). A modified Gompertz model was used for disease-time relationships and provided information on the rate of infection incidence progression (r(g)) and time delayed to start of the incidence progress curve (t(0)). Values of r(g) ranged from near 0 to 1.90, and t(0) varied from 1.3 to more than 10 days. The more aggressive strains showed high r(g), low ED(50) values, and short t(0), whereas the less aggressive strains showed low r(g), high ED(50), and long t (0). The aggressiveness was dependent on plant material type and pear cultivars and was significantly different between strains of E. amylovora. Infectivity titration and kinetic analysis of progression of incidence of infections using the immature pear test and a standardized scale are proposed for assessment of strain aggressiveness. The implications of r(g), ED(50), and t(0) for the epidemiology and management of fire blight are discussed, particularly the wide range of aggressiveness among strains, the degree of host specificity observed in pear isolates, the very high infective potential of this pathogen, the independent action of pathogen cells during infection, and the possible advantage of including aggressiveness parameters into fire blight risk forecasting systems.     

Evidence for age- and size-mediated controls of tree growth from grafting studies

Vegetative propagation techniques such as grafting can be used, in conjunction with field studies, to decouple the relative effects of age and size on tree metabolism and growth. Despite interest in this approach, little attention has been paid to the best metrics for assessing the growth performance of grafted plants over time. Based on an analysis of the grafting literature and our own data, we show that the choice of metrics to assess tree growth can entirely change the conclusions reached about the relative importance of age versus size. We recommend that absolute as well as relative rates of growth are calculated and that scion size be standardized as much as possible at the start of the experiment. Once proper metrics are chosen, all of the available published evidence is largely concordant with two concepts: (1) age-mediated controls of tree growth are likely to be important during the first few years of a tree's life (before phase change); and (2) after the first few years of a tree's life, size-mediated factors largely prevail over age-mediated factors in determining tree growth rates. We found no support for theories invoking age-mediated sink limitations in old trees.