Cumulus Oophorus Expansion of Bovine Oocytes Cultured in vitro: A SEM and TEM Study

Bovine cumulus-oocyte complexes (COCs) were aspirated from antral follicles (3–6 mm in diameter). COCs with a compact multilayered cumulus investment were cultured for 9, 13 and 24 hours, respectively, and processed for scanning or transmission electron microscopy after different periods of culture including a 0 hour control group. In all 0 hour control COCs initial formation of cumulus cell projections, blebs and microvilli were observed. The start of cumulus-oocyte disconnection in cattle was observed after 9 hours of in vitro culture. These connections were almost completely discrupted after 13 hours, when a continuous production of extracellular matrix was observed. The full expansion of corona radiata cells was not observed even after 24 hours. Some cumulus oophorus cells were bound together with junctions of the zonula adherens type after 24 hours when extracellular matrix was found only in deeper layers. The full expansion of corona radiata cells was not the prerequisite for disconnection of cumulus-oocyte complex. Inhalt: Kumulus oophorus Expansion von in vitro kultivierten Rinderoozyten: Eine SEM und TEM Untersuchung Kumulus-Oozyten-Komplexe (COC) von Rindern wurden aus antralen Follikeln aspiriert (Durchmesser 3–6 mm). COC's mit kompakten mehrschichtigen Kumulus wurden für 9, 13 und 24 Stunden kultiviert und für die Scanner- oder Transmissionselektronenmikroskopie vorbereitet. Verschiedene Kulturzeiten incl. einer 0-Stundengruppe als Kontrolle wurden untersucht. In allen Kontroll COC's wurden beginnende Formationen yon Kumuluszellprojektionen, Bläschen und Mikrovilli beobachtet. Nach 9-stündiger in vitro Kultur begann die Lösung zwischen Kumulus und Oozyte. Die Verbindungen waren nach 13 Stunden vollständig unterbrochen, wobei eine kontinuierliche Produktion extrazellulärer Matrix beobachtet wurde. Eine vollständige Expansion der Corona radiata Zellen wurde auch nicht nach 24 Stunden beobachtet. Nach 24 Stunden waren einige Cumulus Oophorus Zellen über Verbindungen des Zonula adherens Typs verknüpft, wobei extrazelluläre Matrix nur in den tieferen Zellschichten gefunden wurde. Eine vollständige Kumulusexpansion ist keine Voraussetzung für die Auflösung des Kumulus-Oozyten-Komplexes.     

Fertilizing capacity of the ejaculate of nutria (Myocastor coypus) after the removal of the seminal vesicles as evaluated by the penetration test and natural mating

The fertility of male coypu sperm following seminal vesicle extirpation was investigated using the penetration test into the egg of Syrian golden hamster (Mesocricetus auratus). Ejaculates were obtained from five males by means of electro-ejaculation under halothane narcosis. The results of the zona-free hamster eggs (ZFHE) penetration test showed that the ejaculates of all the surgically treated coypu males were fertile and that ZFHE value fluctuated from 54 to 76.6%. The results obtained in experiments with natural mating revealed that the extirpation of male coypu seminal vesicles did not affect their fertility. In total 47 foetuses were found post mortem in ten coypu females covered by surgically treated males, which on average represented 4.7 foetuses per female.     

Effect of dietary Schizochytrium microalga oil on selected markers of low‐grade inflammation in rats

The objective of the present study was to evaluate a potential of Schizochytrium microalga oil to alleviate possible negative effects of high‐fat‐high‐energy diets. Forty adult male rats (Wistar Albino) were fed 7 weeks the diet containing beef tallow + evaporated sweetened milk (diet T) intended to cause mild obesity and low‐grade systemic inflammation. Consequently, the animals were divided into four groups by 10 animals each and fed either the T‐diet (control) or the diet containing 6% of safflower oil (S), 6% of fish oil (F) and 6% of Schizochytrium microalga oil (A), respectively, for another 7 weeks. The A‐diet decreased (p < 0.05) live weight to 86% and glycaemia to 85% of control, respectively; an effect of the S‐ and F‐diet on these markers was insignificant (p > 0.05). In comparison with control, higher (p < 0.05) deposition of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) in the epididymal adipose tissue (EAT) of the A‐rats correlated with increased (p < 0.05) plasma adiponectin concentration, but it was without the effect (p > 0.05) on cellular adiponectin content in the EAT. Higher (p < 0.05) EPA+DHA deposition in the liver of the A‐rats correlated with higher expression (149% of control; p < 0.05) of the gene coding for peroxisome proliferator‐activated receptor gamma, and with lower expression (82% and 66%; p < 0.05) of the genes coding for adiponectin receptors AdipoR1 and AdipoR2; no relationship to the expression of receptor GPR120 was found. The A‐diet did not affect amount of the nuclear fraction of the nuclear factor kappa B in the liver, but increased plasma level of anti‐inflammatory cytokine TGF‐β1 (p < 0.05). The presented data agree with results of other in vivo rodent and human studies, but not with literature data regarding in vitro experiments: it can be concluded that the effects of dietary oils on inflammatory markers need further investigation.     

Antimicrobial susceptibility of Escherichia coli from swine, horses, dogs and cats as determined in the BfT-GermVet monitoring program 2004-2006

A total of 417 isolates of Escherichia coli collected from five animal species/organ system combinations from swine [urinary/genital tract (UGT) incl. mastitis metritis agalactia syndrome], horses [genital tract (GT)] and dogs/cats [respiratory tract (RT), UGT and gastrointestinal tract (GIT)] were analysed quantitatively for their susceptibility against different antimicrobial agents by determination of minimum inhibitory concentrations. Regardless of which animal species the strains originated from, resistance appeared most frequently against sulfamethoxazole (18-59%), tetracycline (14-54 %), and ampicillin (14-39%). High percentages of intermediate isolates were observed for cephalothin (39-46 %). In general, low prevalences of resistance were detected for amoxicillin/clavulanic acid (1-4%), gentamicin (1-9%), and cefazolin (0-11%). Generally speaking, the antimicrobial resistance situation among E. coli isolates from horses and small animals is relatively good.     

Modification of essential fatty acid composition in broodstock of cultured European eel Anguilla anguilla L

Farmed eels had lower levels of arachidonic acid (20:4 n‐6) (ARA) and higher ratios of eicosapentaenoic acid (20:5 n‐3) (EPA):ARA compared to wild European eels collected from the Baltic Sea and southern Norwegian coast. Eels fed a formulated feed (JD) with a distribution of essential fatty acids (EFA) resembling wild European eel were sampled after 0, 5, 10, 14 and 44 weeks of feeding to examine changes in fatty acid composition (FAC) in ovaries, visceral fat and muscle. The results showed a slow but steady incorporation of EFA. Lipids are incorporated in the oocytes early in oogenesis, and the leading cohort of oocytes is rich in lipid droplets before the onset of vitellogenesis. This indicates that feeding with optimized broodstock feeds should start early to allow the incorporation of EFA in the first cohort of oocytes. At least 14 weeks of feeding is required to change lipid EFA in broodstock eel to resemble EFA in the diet or in wild fish. After 44 weeks of feeding, ARA was significantly higher in the neutral lipids of ovaries (1.9%) compared to visceral fat (1.2%) or muscle (1.0%). EPA:ARA ratios decreased two‐ to threefold in all tissues examined during that time. ARA and docosahexaenoic acid (22:6 n‐3) (DHA) had accumulated in ovarian polar lipids.     

Fertilizer ammonium : nitrate ratios determine phosphorus uptake by young maize plants

We investigated the interacting effects of inorganic nitrogen and the main inorganic phosphorus form in dairy manure (dicalcium phosphate, CaHPO₄) on growth, nutrient uptake, and rhizosphere pH of young maize plants. In a pot experiment, three levels of CaHPO₄ (0, 167, and 500 mg P pot^?1) were combined with nitrogen (637 mg N pot^?1) applied at five NH₄‐N : NO₃‐N ratios (0 : 100, 25 : 75, 50 : 50, 75 : 25, and 100 : 0) and a nitrification inhibitor in a concentrated layer of a typical acid sandy soil from Denmark. ¹⁵N‐labeled NH₄‐N was applied to differentiate the role of nitrification and to partition nitrogen uptake derived from NH₄‐N. Among treatments including nitrogen, shoot biomass, rooting and phosphorus uptake were significantly higher at the five‐leaf stage when CaHPO₄ was applied with NH₄‐N : NO₃‐N ratios of 50 : 50 and 75 : 25. In these treatments, rhizosphere pH dropped significantly in direct proportion with NH₄‐N uptake. The fertilizers in the concentrated layer had a root‐inhibiting effect in treatments without phosphorus supply and in treatments with pure NO₃‐N or NH₄‐N supply. Increased nitrogen uptake as NH₄‐N instead of NO₃‐N reduced rhizosphere pH and enhanced acquisition of applied CaHPO₄ by young maize plants, which may have positive implications for the enhanced utilization of manure phosphorus.     

Recovery of pigments from Origanum majorana L. by extraction with supercritical carbon dioxide

Extraction of pigments (chlorophylls and carotenoids) from marjoram (Origanum majorana L.) with supercritical carbon dioxide was investigated. The aim of this study was to map the effects of extraction pressure and temperature on the yield of coloring materials by applying a 3(2) full factorial design with three repeated tests in the center of the design. For comparison, laboratory and pilot plant Soxhlet extractions were carried out using ethanol and n-hexane solvents. The compositions of pigments in marjoram extracts were determined by HPLC. Similar amounts of carotenoids, in addition to 40% of chlorophylls and their derivatives, were recovered from the supercritical fluid extraction, in comparison to the ethanol Soxhlet extraction.