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.     

Composition and biological activity of essential oils against Metopolophium dirhodum (Hemiptera: Aphididae) cereal crop pest

BACKGROUND: Natural pesticides based on plant essential oils may represent alternative crop protectants. This study analysed the chemical constituents and bioactivities of essential oils from Schinus areira L., Rosmarinus officinalis L. and Tagetes terniflora Kunth against winged and wingless adults of Metopolophium dirhodum (Walk.). RESULTS: The major component of S. areira was α‐phellandrene, that of R. officinalis was 1,8‐cineole and that of T. terniflora was cis‐ocimene. Rosemary essential oil was more effective than the others in immersion method tests. In contact toxicity, the rosemary oil was the most toxic to wingless adults and the S. areira leaf oil was the most toxic to winged adults. All the EOs produced some degrees of repellency in adults and effects on the reproduction and demographic parameters. CONCLUSION: These results showed that the essential oils from S. areira, R. officinalis and T. terniflora could be used as an alternative in the management of M. dirhodum. Copyright © 2012 Society of Chemical Industry     

Differential interactions of virulent and non-virulent H. parasuis strains with na?ve or swine influenza virus pre-infected dendritic cells

Pigs possess a microbiota in the upper respiratory tract that includes Haemophilus parasuis. Pigs are also considered the reservoir of influenza viruses and infection with this virus commonly results in increased impact of bacterial infections, including those by H. parasuis. However, the mechanisms involved in host innate responses towards H. parasuis and their implications in a co-infection with influenza virus are unknown. Therefore, the ability of a non-virulent H. parasuis serovar 3 (SW114) and a virulent serovar 5 (Nagasaki) strains to interact with porcine bone marrow dendritic cells (poBMDC) and their modulation in a co-infection with swine influenza virus (SwIV) H3N2 was examined. At 1 hour post infection (hpi), SW114 interaction with poBMDC was higher than that of Nagasaki, while at 8 hpi both strains showed similar levels of interaction. The co-infection with H3N2 SwIV and either SW114 or Nagasaki induced higher levels of IL-1β, TNF-α, IL-6, IL-12 and IL-10 compared to mock or H3N2 SwIV infection alone. Moreover, IL-12 and IFN-α secretion differentially increased in cells co-infected with H3N2 SwIV and Nagasaki. These results pave the way for understanding the differences in the interaction of non-virulent and virulent strains of H. parasuis with the swine immune system and their modulation in a viral co-infection.     

Post-fire salvage logging increases restoration costs in a Mediterranean mountain ecosystem

Post-fire salvage logging (i.e. felling and removing burnt trees, often eliminating the remaining woody debris) is a practice routinely performed by forest managers worldwide. In Mediterranean-type ecosystems, salvage logging is considered a measure to reduce future reforestation costs, but this assumption remains largely untested. We made a cost analysis of different management schemes, addressing the immediate post-fire burnt-wood management as well as the costs and success of subsequent reforestation efforts. Two experimental 25-ha plots were established in a burnt pine reforestation of SE Spain, in which three replicates of three post-fire treatments were applied: non-intervention (NI), partial cut plus lopping (PCL; felling and lopping off the branches from most of the trees, leaving all biomass in situ), and salvage logging (SL). After 4?years, a mechanised reforestation was undertaken, and seedling mortality was monitored for 2?years. The cost of all management operations was recorded in situ, and the cost of re-planting the dead seedlings was estimated according to the expenses of previous reforestation. Initial cost of wood management was greatest in SL and zero in NI. Reforestation cost was highest in NI and lowest in SL, and seedling-mortality rates proved lowest in PCL (43?% vs. 51?% and 52?% in SL and NI, respectively). Considering all the post-fire management operations, salvage logging did not provide particular economic advantages for forest restoration, and had an overall cost of 3,436?±?340?€/ha. By contrast, NI and PCL reduced total restoration costs by 50 and 35?%, respectively, and PCL indeed promoted restoration success. We suggest that the full cost of management operations needs to be considered when evaluating the economic implications of post-fire salvage logging.     

Distribution and persistence of atypical porcine pestivirus in experimentally inoculated pigs

Atypical porcine pestivirus (APPV) is a cause of congenital tremors (CTs) in piglets and has been found in swine populations around the globe. Although systemic distribution of the virus has been reported, there is limited information regarding viral localization at the cellular level and distribution at the tissue level. We collected multiple tissues from 2-d-old piglets (n = 36) born to sows inoculated at 45 or 62 d of gestation with APPV via 3 simultaneous routes: intravenous, intranasal, and directly in amniotic vesicles. In addition, 2 boars from APPV-inoculated sows with CT were raised and euthanized when 11 mo old. In situ hybridization performed on tissue samples from piglets demonstrated a broad and systemic distribution of viral RNA including endothelial cells, fibroblasts, and smooth muscle. Labeling in tissues was more pronounced in piglet tissues compared to boars, with the notable exception of diffuse labeling of the cerebellum in boars. Presence of APPV in boar tissues well after resolution of clinical signs suggests persistence of APPV similar to other pestiviruses.     

Essential oils for preventative treatment and control of Asian soybean rust

The objective of the work was to evaluate the effect of essential oils as a preventive treatment to control Asian soybean rust. Initially the fungitoxic effect of the essential oils of Hyptis marrubioides, Aloysia gratissima and Cordia verbenacea was tested on the urediniospores of Phakopsora pachyrhizi, through in vitro tests. The in vivo test was set up in a greenhouse, using cultivar MGBR-46. The treatments consisted of the three oils at different concentrations, a fungicide based on pyraclostrobin + epoxyconazole and a control. To verify the potential of the essential oils in preventive treatment, inoculation was conducted at 0, 6, 12 and 24?h intervals, after application of the treatments. All treatments inhibited 100% of Phakopsora pachyrhizi germination. In the in vivo test, it was observed that all of the oils were efficient in controlling soybean rust, by preventive treatment, mainly at the higher concentrations. Efficiency of the oils was reduced with the increase of the interval between the application of the treatments and the onset of the pathogen. The essential oils, at the tested dosages, were not as efficient as the pyraclostrobin + epoxyconazole based fungicide.     

Cross‐Sectional Study of the Prevalence of Radiographic Degenerative Joint Disease in Domesticated Cats

Objective: To determine the prevalence of radiographic signs of degenerative joint disease (DJD) in a randomly selected sample of domestic cats. Study Design: Prospective observational study. Animals: Client‐owned cats. Methods: Cats (n=100) from a single practice and equally distributed across 4 age groups (0–5; 5–10; 10–15, and 15–20 years old) were randomly selected (regardless of heath status) and sedated for orthogonal radiographic projections of all joints and the spine. Quasi‐Poisson regression analysis was used to investigate the relationship between patient demographics, blood biochemistry, hematologic and urine analysis variables, and DJD severity. Results: Most (92%) cats had radiographic evidence of DJD; 91% had at least 1 site of appendicular DJD and 55% had ≥1 site of axial column DJD. Affected joints in descending order of frequency were hip, stifle, tarsus, and elbow. The thoracic segment of the spine was more frequently affected than the lumbosacral segment. Although many variables were significantly associated with DJD, when variables were combined, only the association between age and DJD was significant (P<.0001). For each 1‐year increase in cat age, the expected total DJD score increases by an estimated 13.6% (95% confidence interval: 10.6%, 16.8%). Conclusion: Radiographically visible DJD is very common in domesticated cats, even in young animals and is strongly associated with age. Clinical Relevance: DJD is a common disease of domesticated cats that requires further investigation of its associated clinical signs.     

Use of quantitative two-dimensional color tissue Doppler imaging for assessment of left ventricular radial and longitudinal myocardial velocities in dogs

OBJECTIVE: To determine left ventricular free wall (LVFW) radial and longitudinal myocardial contraction velocities in healthy dogs via quantitative 2-dimensional color tissue Doppler imaging (TDI). ANIMALS: 100 dogs. PROCEDURE: TDI was used by a single trained observer to measure radial and longitudinal myocardial movement in the LVFW. Radial myocardial velocities were recorded in segments in the endocardial and epicardial layers of the LVFW, and longitudinal velocities were recorded in segments at 3 levels (basal, middle, apical) of the LVFW. RESULTS: LVFW velocities were higher in the endocardial layers than in the epicardial layers. Left ventricular free wall velocities were higher in the basal segments than in the middle and apical segments. Radial myocardial velocity gradients, defined as the difference between endocardial and epicardial velocities, were (mean +/- SD) 2.5 +/- 0.8 cm/s, 3.8 +/- 1.5 cm/s, and 2.3 +/- 0.9 cm/s in systole, early diastole, and late diastole, respectively. Longitudinal myocardial velocity gradients, defined as the difference between basal and apical velocities, were 5.9 +/- 2.2 cm/s, 6.9 +/- 2.5 cm/s, and 4.9 +/- 1.7 cm/s in systole, early diastole, and late diastole, respectively. A breed effect was detected for several systolic and diastolic TDI variables. In all segments, systolic velocities were independent of fractional shortening. CONCLUSIONS AND CLINICAL RELEVANCE: LVFW myocardial velocities decreased from the endocardium to the epicardium and from base to apex, thus revealing intramyocardial radial and longitudinal velocity gradients. These indices could enhance conventional echocardiographic analysis of left ventricular function in dogs. Breed-specific reference intervals should be defined.