Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle

The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus–oocyte complexes) cell‐conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus–oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real‐time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell‐specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K‐Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian‐related diseases and/or the identification of biomarkers for oocyte and embryo quality.     

Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.     

The serological responses of chickens to mass vaccination with a live V4 Newcastle disease virus vaccine in the field and in the laboratory 2. Layer pullets

Layer chickens on a commercial started pullet farm were vaccinated once at 31 to 52 days of age by drinking water or aerosol with live V4 Newcastle disease virus (NDV) vaccine. Flockmates which had been rehoused in laboratory isolation pens shortly beforehand were similarly vaccinated. Samples of birds were bled at intervals and the serums tested for haemagglutination inhibiting antibody to NDV. Log2 mean titres of up to 4.88 and assumed protection levels (based on the percentage of birds with log2 titres of 4 or greater) of up to 81%, were obtained in the field trials within 4 weeks of vaccination. A subsequent laboratory trial further compared the response of different breeds of chicken to different routes of vaccination. Differences were observed between breeds, routes of vaccination, and parallel field and laboratory trials. The results show that this V4 vaccine can produce an adequate serological response following mass vaccination of Australian layer pullets housed under commercial conditions, and that care should be exercised in extrapolating results obtained under laboratory conditions.     

Seroprevalence of psittacine beak and feather disease in wild psittacine birds in New South Wales

SUMMARY A haemagglutination inhibition assay was used to detect antibody to psittacine beak and feather disease virus in sera from wild sulphur crested cockatoos (Cacatua galerita), galahs (Eolophus roseicapillus), short-billed corellas (Cacatua sanguinea), eastern long-billed corellas (Cacatua tenuirostris) and other psittacine birds in New South Wales. The seroprevalence of psittacine beak and feather disease ranged from 41% to 94% in different flocks, indicating infection with the virus is widespread in wild populations.     

Plasma indicators of muscle damage in a model of nutritional myopathy in weaner sheep

SUMMARY Subclinical nutritional myopathy was induced in 5-month-old sheep by feeding them a diet low in vitamin E and selenium. Subsequently clinical myopathy was induced by dosing with protected polyunsaturated fatty acids. Plasma activities of creatine kinase (CK), pyruvate kinase, aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase and aldolase, enzymes of muscle origin, all remained above their reference ranges in clinically affected sheep, but fluctuated widely. Similar fluctuations occurred in subclinically affected animals, resulting in some activities being within the reference ranges and some above, at different times. Plasma malondialdehyde, an indicator of lipid peroxidation, proved of no diagnostic value. Terminal plasma CK activities were significantly correlated with microscopic damage in the vastus lateralis (VL), but not the vastus intermedius (VI) or the tensor fascia lata (TFL) muscles. AST was the most highly correlated with damage in VI and VL. In two clinically affected sheep successfully treated with an oral dose of α-tocopherol actetate all enzymes decreased steadily to within their reference ranges, at rates probably related to their plasma half-lives. These results suggest that measurement of plasma CK activity would be useful in monitoring recovery of treated sheep.     

Continental drainage in north america during the phanerozoic from Nd isotopes

Neodymium (Nd) isotopic data show consistent patterns in the sources of sedimentary rocks in North America at a continental scale. Between 600 and 450 million years ago (Ma), ancient continental shield sources dominated. Around 450 Ma, detritus from the Caledonian-Appalachian mountains overwhelmed sediment from all older sources, and is documented over large areas of the southern, western, and northern margins of North America. This material continued to dominate the sediment supply until about 150 Ma, probably due to cannibalistic recycling of sedimentary rocks formed earlier. Around 150 Ma, the rising western Cordillera delivered new and different detritus to the sedimentary system.