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.     

The Effect of Lead and Zinc on Plant Growth and Chlorophyll Content of Holcus lanatus L.

Root length of Holcus lanatus L. declined rapidly with increasing lead and zinc concentrations in nutrient solution. In all used Pb and Zn concentrations root growth of genotypes coming from a Pb-Zn mine area, was greater than that of the control, suggesting that these genotypes evolved tolerance to Pb and Zn.
Negative correlation was also observed between chlorophyll content and increased heavy metal concentrations. The greater chlorophyll content found in tolerant genotypes, in different Pb and Zn concentrations, in comparison with the control, could be served to distinguish tolerant and non-tolerant genotypes.     

Physiological Analysis of Yield Variation in Mungbean (Vigna radiata [Roxb.] Wilczek)

Two mungbean varieties, Guj-1 and PIMS-1 differing in their productivity potential, were examined to investigate their photosynthetic system at various stages of growth in relation to yield with two different dates of sowing. Vertical leaves were found to be beneficial. On the other hand, excessive leaf area during the later stages of growth may be detrimental to yield. Lower LAD at seedling stage and overall high NAR values may be reflected in higher grain yield. NAR declined as season progressed and this, in turn, may be related with increase in LAI. The grain yield was considerably more at the first date of sowing as compared to second date of sowing, the effect was being more pronounced in var. PIMS-1 . Various environmental factors recorded at two different dates of sowing and at various stages of growth indicated that these factors may have influenced the yield potential at two different dates of sowing.     

Dynamics of the Cumulation of Macroelements – Phosphorus, Potassium, Calcium and Magnesium During Development to Maturity of Soybean Seed

Dynamics of the cumulation of phosphorus, potassium, calcium and magnesium in developing seed of four soybean genotypes under the defined climatic conditions in the course of three years was observed. Samples were being taken at weekly intervals from the point at which the grain reached a size suitable to be analyzed up to the full maturity of the seed. By analyzing the results it was found out that the intense cumulation of the minerals tested took place during the stages III and IV of the seed development, that is, in the course of intense cumulation of dry matter and the most expressive metabolic activity in the seed. Investigation of the impact of climatic conditions provides evidence that cumulation of the tested macroelements has been favourably influenced by increased precipitation along with moderate air temperatures within the seed developing period. The highest content of the minerals found with the cultivar Maksimirka independently of the year investigated reveals that mineral cumulation in soybean seed is affected by the cultivar and genetic particularity.     

Comparison of Three Solvent Systems for Extraction of Chlorophyll a from Fish Pond Phytoplankton Communities

Chloroform-methanol (2:1 v/v), absolute methanol, and 90% acetone were evaluated for their effectiveness as extractants of chlorophyll a from samples of phytoplankton communities collected from catfish ponds. Chloroform-methanol consistently extracted more chlorophyll a than either 90% acetone or methanol. Precision for the methanol extraction was also unacceptably low, with an average coefficient of variation of 17%. Average coefficients of variation for the chloroform-methanol and 90% acetone extraction procedures were 6 and 5%, respectively. Filtered samples should be steeped in chloroform-methanol for at least 4 h to obtain maximum chlorophyll extraction, and the addition of MgCO₃ to the extractant as a buffer is not necessary.     

Changes in several blood and urine parameters during combined hyperfluid therapy for the treatment of chronic obstructive bronchitis (COB) in the horse

During the combination of oral and intravenous application of saline solution for treatment of the COPD of horses the level of hydraemia basing on the total protein concentration in the serum, the urine production and the specific weight of urine was determined. Additionally the development of serum concentration and of renal excretion rates of potassium, calcium and magnesium were ascertained. The level of hydraemia resulting from the combined method is almost identical with the solely intravenous performed hyperinfusion therapy. Due to the excessive application of fluid an extremely high level of urine production is reached which causes a reduced specific weight as well as an increased renal excretion of potassium, calcium and magnesium. The result is a real loss of electrolytes which is - apart from hydraemia - the reason for the lower level of the corresponding serum concentrations. This seems to be important, especially for potassium and magnesium, because the organism is unable to compensate the loss of these electrolytes in the same way as the loss of calcium. In connection with the renal loss of electrolytes during the high level of urine production glucosuria is observed.     

Microcirculation in the follicles and corpora lutea of the bovine ovary--a corrosion anatomical study

With the help of a new synthetic material follicles and corpora lutea from the ovaries of 80 cows in different stages of the sexual cycle were examined for modifications of their blood vessel architecture. In the theca externa the main artery which runs towards the follicle, separates into arterioles of which capillaries derive in the theca interna, building a basket-like network around the zona granulosa. After ovulation the follicle wall forms characteristic fields. Capillaries grow into the zona granulosa and thus the folds form the lobular structure of the corpus luteum. Capillaries which rise from the ruptured part of the follicle, spread centripetally and together with lutein cells they build up the not lobulated cupola of the corpus luteum. The regression of the corpus luteum starts around the 16th day of the cycle, beginning at the capillary network of the cupola. A contraction of the vessels and a quick decrease of the volume follow.     

The effect of hypervitaminosis D3 on the bones of the chicken. 2. Electron microscopic studies

In an experiment of 36 days duration 46 one-day-old chicks were divided into 5 groups and fed with different concentrations of vitamin D3. The animals of the group which lacked vitamin D3, showed the typical rachitic lesions. After a 15 days lack of vitamin D3 the chicks of another group were treated with standard food (2000 I.U. vitamin D3/kg food) with the consequence of an approximation of the analyzed parameters to those of the control group within 3 weeks. When fed with 60,000 I.U. of vitamin D3 after a 15 days lack of this vitamin, the animals showed an over-hasty healing process, ending up with signs of intoxication which were even more conspicuous when fed with 120,000 I.U. of vitamin D3. Besides an increasing calcification of osteoblasts and endothelial cell membranes as well as a degeneration of osteoblasts, a clear increase of eosinophilic granulocytes could be noticed. In all groups free erythrocytes within the ground substance were found. There was no evidence of necroses of osteocytes or of bone.     

In situ radiometric mapping of soil erosion and field-moist bulk density on cultivated fields

Abstract. Recent developments in in situγ ray spectrometry offer a new approach to measuring the activity of radionuclides such as ¹³⁷Cs and ⁴⁰K in soils, and thus estimating erosion or deposition rates and field moist bulk density (ρ_m). Such estimates would be rapid and involve minimal site disturbance, especially important where archaeological remains are present. This paper presents the results of a pilot investigation of an eroded field in Scotland in which a portable hyper pure germanium (HPGe) detector was used to measure γ ray spectra in situ. The gamma (γ) photon flux observed at the soil surface is a function of the ¹³⁷Cs inventory, its depth distribution characteristics and ρ_m. A coefficient, Q_Cs, derived from the forward scattering of ¹³⁷Cs γ ray photons within the soil profile relative to the ¹³⁷Cs full energy peak (662 keV), was used to correct the in situ calibration for changes in the ¹³⁷Cs vertical distribution in the ploughed field, a function of tillage, soil accumulation and ρ_m. Based on only 8 measurements, the agreement between in situγ ray spectrometry and soil sample measurements of ¹³⁷Cs inventories improved from a non significant r²=0.05 to a significant r²=0.62 (P<0.05). Erosion and deposition rates calculated from the corrected in situ¹³⁷Cs measurements had a similarly good agreement with those calculated from soil cores. Mean soil bulk density was also calculated using a separate coefficient, Q_K, derived from the forward scattering γ photons from ⁴⁰K within the soil relative to the ⁴⁰K full energy peak (1460 keV). Again there was good agreement with soil core measurements (r²=0.64; P<0.05). The precision of the in situ¹³⁷Cs measurement was limited by the precision with which Q_Cs can be estimated, a function of the low ¹³⁷Cs deposition levels associated with the weapons testing fallout and relatively low detector efficiency (35%). In contrast, the precision of the in situ ρ_m determination was only limited by the spatial variability associated with soil sampling.     

Comparison of partial ribosomal DNA sequences of Babesia gibsoni occurring in Miyazaki prefecture, Japan.

Nucleotide sequences of ribosomal DNA (rDNA) of Babesia (B.) gibsoni occurring in Miyazaki, western Japan, were examined using blood samples obtained from seven dogs suffering from natural canine babesiosis. DNA isolated from these blood samples was subjected to the polymerase chain reaction (PCR). The nucleotide sequences of the PCR products were determined and compared with other rDNA sequences of B. gibsoni isolated from Asia, Europe and U.S.A. Although homology values between our isolates and those isolated from Europe and U.S.A. were both 84.0%, respectively, our isolates were identical to the Asian types. In conclusion, B. gibsoni occurring in Miyazaki was revealed to have the genotype Asia 1 or Asia 2 from a comparison of the partial rDNA sequences.