Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Electron microscope observations of Ceratocystis ulmi-induced tylosis development in Ulmus1

Electron microscope observations of Ceratocystis ulmi-induced tylosis development in 3 species of elm were made during the first 2 weeks after artificial inoculation. Tylosis forming from both ray and vasicentric parenchyma occluded vessels adjacent to those containing the fungus conidia within one day after inoculation. The expanding protoplast forced the intact vessel-parenchyma “pit membrane” and protective layer through the pit into the vessel lumen. Cell organization during this expansion is discussed. The observations on tylosis development for the 3 elms used were identical.     

Follow-up of mercury levels in fish,human hair and urine in the Madeira and Tapajós basins,Amazon, Brazil

Informal economy of gold mining has contaminated some important river basins in Amazon. Follow-up studies on critical compartments showed some areas with high Hg levels in fish as well as in human hair samples. Average Hg in piscivorous fish in the Madeira river itself was 846 ppb (N=284) with a maximum of 3921 ppb. Mercury in fish from non polluted areas in this basin shows high variability, even for single species. A seasonal variation in Hg content was observed, with higher values at the end of the dry season. In the upper Tapajós basin comparable values were found for fish but with a definite decreasing trend downstream. Average value for piscivorous fish in the whole Tapajós basin is 482 ppb (N=122) with a maximum value of 3770 ppb. Hair Hg was higher in fishing villages in the Tapajós (average: 17 ppm; with N=432 and maximum value of 176 ppm) than in the Madeira (average: 9 ppm; N=169; maximum 71 ppm), and data from some areas of the Tapajós suggest a decrease with time. Mercury was much higher in urine of goldshop workers in Santarém (low Tapajós) than in Alta Floresta (high Tapajbs) and show a decreasing trend in both cases, probably related to the significant decline in gold mining activities during the study period (1986–1994).     

The effects of different maternal dietary iodine concentrations on Japanese quail II. Thyroid function in embryos and hatchlingsi

Embryos (day I4 of the 16.5 day incubation period) and I day old chicks of Japanese quail (Coturnix japonica) were used to study the effects of egg I on thyroid development. The maternal diets were supplemented with 0–1200 μg I/kg of purified diet. Thyroid function, of embryos and chicks (before feeding), adapted to a wide range of egg I availability.We measured body weights, thyroid weights, thyroidal ¹²⁵I uptakes, stable I concentrations in thyroid glands and triiodothyronine (T₃) and thyroxine (T₄) concentrations in the serum.With low egg I (maternal diets with <50 μg I/kg) embryonic and chick thyroid glands were hypertrophied, thyroidal I content was very low and thyroidal radioiodine uptakes were high. Serum hormone concentrations were not significantly different from those of embryos and chicks from eggs with higher I contents. Thyroid weights were lowest in young from eggs of hens on the 150 μg I/kg diet, and the other variables studied also indicated this maternal diet provided sufficient egg I for the thyroid function of embryos and hatchlings.With high egg I (maternal diets with 300–1200 μg I/kg) thyroidal radioiodine uptakes were low but thyroidal I incorporation was related to egg I content. Serum hormone concentrations were not significantly altered by high I availability except that serum T₄ concentrations differed in a comparison of the extremes (0 vs 1200 μg I/kg in the maternal diet). As expected, from the consistency of serum hormone concentrations, embryonic growth was not affected by the I content of the eggs.In agreement with earlier studies on quail hens, our results indicate that feed supplementation of 150 μg I/kg is sufficient to meet needs without requiring alterations in thyroid function to maintain serum hormone concentrations. Developing thyroid glands adapt effectively to egg I concentrations of 0.2 to 4.1 μg I/g yolk which result from maternal diets with 0–1200 μg I/kg.     

Detection of feline coronavirus infection in captive cheetahs (Acinonyx jubatus) by polymerase chain reaction.

Feline coronavirus genetic elements were detected by polymerase chain reaction from blood, fecal samples, and effusive fluid collected from 33 cheetahs in the U.S.A. Feline coronavirus-specific serum antibodies were also measured by indirect immunofluorescence. Ten cheetahs were positive for viral shedding by polymerase chain reaction, whereas 13 were seropositive by immunofluorescence. Results of serology did not consistently correlate with shedding of virus, and the capture antigen used for detection of feline coronavirus-specific antibodies had a significant impact on results. Testing of samples from one population over a 1-yr period indicated chronic infection in some animals. These relatively healthy carrier animals were a source of virus for contact animals. Screening programs in cheetah populations for feline coronavirus infection may be most reliable if a combination of serologic analysis and viral detection by polymerase chain reaction is used.     

Valine partitioning and kinetics between the gastrointestinal tract and hind limbs in lambs with an adult Trichostrongylus colubriformis burden

Intestinal parasitic infection increases the demand for AA because of increased protein synthesis in the intestine and increased luminal losses of AA, and these increased demands may be supported by increased mobilization of AA from the skeletal muscles. Two experiments were conducted to determine the effects of parasitic infection on valine kinetics within the gastrointestinal tract and hind limbs of lambs fed fresh forages. On d 1, lambs were given 6,000 stage-3 Trichostrongylus colubriformis larvae per day for 6 d (n = 6) or kept as parasite-free controls (n = 6) and fed fresh lucerne (Medicago sativa; Exp. 1) or fresh sulla (Hedysarum coronarium; Exp. 2). On d 48, valine kinetics within the mesenteric- (MDV) and portal-drained viscera (PDV) and hind limbs were obtained by carrying out concurrent infusions of para-amminohippuric acid into the mesenteric vein and indocyanin green into the abdominal aorta (for blood flow), and [3,4-(3)H]valine into the jugular vein and [1-(13)C]valine into the abomasum for 8 h (for kinetics). During the infusions, blood was collected from the mesenteric and portal veins and from the mesenteric artery and vena cava, and plasma was harvested. After the 8-h infusion, lambs were euthanized, ileal digesta were collected, and tissues were sampled from the intestine and muscle (biceps femoris). Tissues, digesta, and plasma were analyzed for valine concentration, specific radioactivity, and isotopic enrichment. In both experiments, intestinal worm burdens on d 48 were greater in parasitized lambs (P = 0.0001 and 0.003). In Exp. 1, parasitic infection increased (P = 0.03) the total valine irreversible loss rate (ILR) in the MDV and PDV. In Exp. 2, luminal ILR of valine in the MDV was reduced (P = 0.01); however, ILR of valine in the PDV was unaffected. Despite these changes within the MDV and PDV, parasitic infection did not affect the ILR of valine within the hind limbs, and valine transport rates were largely unchanged. We suggest that the increased mobilization of AA from the hind limbs that might have occurred in the early phase of inflammation was no longer required when the parasitic infection was established. The MDV and PDV data may indicate that the non-MDV parts of the PDV play an important role in this adaptation, which warrants further study.     

Effect of condensed tannins on egg hatching and larval development of Trichostrongylus colubriformis in vitro

The effects of condensed tannins extracted from seven forages on the viability of the eggs and first stage (L1) larvae of the sheep nematode Trichostrongylus colubriformis were evaluated in in vitro assays. The extracts of condensed tannins were obtained from Lotus pedunculatus (LP), Lotus corniculatus (LC), sulla (Hedysarum coronarium), sainfoin (Onobrychus viciifolia), Dorycnium pentaphylum (DP), Dorycnium rectum (DR) and dock (Rumex obtusifolius). Extracts containing 200 to 500 microg/ml reduced the proportion of eggs that hatched. The larval development assay was used to evaluate the effect of the extracts on the development of either eggs or L1 larvae to L3 infective larvae. Development was allowed to proceed for seven days by which time the larvae in control incubations had reached the infective L3 stage. Extracts containing 200 microg/ml from LP, DP, DR or dock prevented egg development, and only 11, 8 and 2 per cent of the eggs developed to L3 larvae with extracts from LC, sulla and sainfoin, respectively. When the concentration was 400 microg/ml no eggs developed to L3 larvae. The addition of the extracts after hatching also inhibited the development of L1 to L3 larvae; 200 microg/ml extracted from LP, LC, sulla, sainfoin, DP, DR and dock resulted in only 14, 18, 17, 15, 14, 16 and 4 per cent of L1 larvae developing to the L3 stage compared with 85 per cent for controls, and 400 microg/ml further reduced the development of L1 larvae. Statistical analyses showed that when the extracts were added before hatching they were significantly (P<0.001) more effective at inhibiting the larval development than when they were added after hatching. The condensed tannins from dock had the greatest inhibitory effect on egg development followed by the tannins from DR, sainfoin, DP, LP, sulla and LC.     

Multiple rotation yields for intensively managed plantations in the Amazon basin

Jari Celulose S. A. is a Brazilian-owned industrial enterprise located along the Jari River (the first major south-flowing tributary of the Amazon). The company produces bleached kraft pulp for international and domestic sales. The raw material of this pulp production comes from the 75,000 ha of intensively managed plantations of Pinus caribaea (Morelet) and Eucalyptus spp. harvested on 10- and 6-year rotations, respectively. All currently active Jari plantations were converted from primary forest between 1968 and 1982. Silvicultural practices have been modified over successive rotations, including plantation species changes, increased intensity of site preparation, fertilization, and chemical weed control. A review of data from the early 1970s to present indicates an increase in productivity over successive rotations and several plantation locations and soil types. The reasons for this improvement are primarily related to the higher levels of silvicultural inputs and dramatic genetic gains. The long-term sustainability of this productivity is discussed in terms of current and future inputs. This revised version was published online in June 2006 with corrections to the Cover Date.     

Organogenesis and foetal haemodynamics during the normal gestation of healthy black‐rumped agoutis (Dasyprocta prymnolopha,Wagler, 1831) bred in captivity

The objective of this study was to define the patterns of organogenesis and foetal haemodynamics during the normal gestation of healthy agoutis (Dasyprocta prymnolopha) kept in captivity. Thirty pregnant agoutis that ranged in size from small to medium and weighed between 2.5 and 3 kg underwent B‐mode and Doppler ultrasonography for the biometric evaluation of the foetal organs. The foetal aortic blood flow proved to be predominantly systolic, and the measured flow velocity was 78.89 ± 2.95 cm/s, with a maximum pressure gradient of 2.12 ± 0.27 mmHg. The liver was characterized by its large volume, occupying the entire cranial aspect of the abdominal cavity, and it was associated cranially with the diaphragm and caudally with the stomach. The flow velocity in the portal vein was estimated to equal 12.17 ± 2.37 cm/s, with a resistivity index of 0.82 ± 0.05. The gallbladder was centrally located and protruded cranially towards the diaphragm. The spleen was visualized as an elongated structure with tapered cranial and caudal extremities, and the foetal kidneys were visualized bilaterally in the retroperitoneal region, with the right kidney positioned slightly more cranially than the left. The morphological characterization and hemodynamic analysis of the foetal organs of black‐rumped agoutis via B‐mode and Doppler ultrasonography allow determination of the vascular network and of reference values for the blood flow required for perfusing the anatomical elements essential for maintaining the viability of foetuses at different gestational ages.     

Major compositional units of the moon: lunar prospector thermal and fast neutrons

Global maps of thermal and fast neutron fluxes from the moon suggest three end-member compositional units. A high thermal and low fast neutron flux unit correlates with the lunar highlands and is consistent with feldspathic rocks. The South Pole-Aitken basin and a strip that surrounds the nearside maria have intermediate thermal and fast neutron flux levels, consistent with more mafic rocks. There appears to be a smooth transition between the most mafic and feldspathic compositions, which correspond to low and high surface altitudes, respectively. The maria show low thermal and high fast neutron fluxes, consistent with basaltic rocks.