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.     

Rhabdomyosarcoma in the Japanese medaka, Oryzia latipes (Temminck & Schlegel) and guppy, Poecilia reticulata Peters

Abstract. Three cases of skeletal muscle neoplasms occurring in two small fish species used in carcinogen tests are reported. The cases illustrated a wide range of histologic patterns, and consisted of a well-differentiated juvenile type, a well-differentiated pleomorphic type and a poorly differentiated pleomorphic type. A rhabdomyosarcoma with juvenile type features developed in the Japanese medaka, Oryzias latipes , exposed to 0·5 ml/1 of the solvent dimethylformamide. A pleomorphic type rhabdomyosarcoma occurred in one medaka and in one guppy, Poecilia reticulata , exposed to the carcinogen methylazoxymethanol acetate. These cases indicate the potential for skeletal muscle cells in medaka and guppy to become neoplastic but, because the tumours occurred at a low frequency, there may not necessarily be a chemical aetiology.     

Predicting lean growth while accounting for correlated traits.

Lean tissue growth rate is usually estimated from indirect measurements including growth rate. A procedure to determine prediction equations for lean tissue growth rate is proposed. The procedure restricts the regression of fat growth rate on predicted lean growth rate to be equal to the regression of fat growth rate on actual lean growth rate. The restriction can be phenotypic or genetic if suitable parameter estimates are available. When applied phenotypically, selection on predicted lean tissue growth rate will result in selection differentials for both fat and lean tissue growth rates that are proportional to those obtained by direct selection for lean tissue growth rate. This restriction is desirable because expected correlated changes in fat are used to justify selection for lean tissue growth. Conventional prediction procedures have ignored correlated changes and obscured the original intent of using lean tissue growth rate as a biological selection criterion. When using conventional procedures to predict a biological selection criterion from indirect measurements, changes in important correlated traits may depend more on the choice of indirect measurements than on the choice of selection criterion.     

The Content of β-endorphin-like Immunoreactivity in Porcine Corpus Luteum and the Potential Roles of Progesterone, Oxytocin and Prolactin in the Regulation of β-endorphin Release from Luteal Cells in vitro

The amount of β‐endorphin‐like immunoreactivity (β‐END‐LI) in porcine corpora lutea from several stages of the oestrous cycle and the effects of progesterone, oxytocin, and prolactin on β‐END‐LI secretion in vitro by luteal cells were studied. Porcine corpora lutea obtained on days 1–5, 6–10, 11–13, 14–18, and 19–21 of the cycle were used to prepare extracts for β‐END‐LI determination. Additionally, corpora lutea from days 11–13 and 14–18 were enzymatically dissociated and isolated luteal cells were used for further study of β‐endorphin secretion in vitro. Cells were cultured in serum‐free defined M 199 medium (10⁶ cells/ml) at 37°C under 5% CO₂ in air, for 12 h. The influences of the following factors on β‐END‐LI secretion by luteal cells were tested: progesterone (10^–9, 10^–7 and 10^–5M ), oxytocin (0.01, 0.1, 1 and 10 ng/ml), and prolactin (0.1, 1, 10 and 100 ng/ml). The β‐END‐LI contents in extracts and media were measured by radioimmunoassay. The tissue concentration of β‐END‐LI was lowest on days 1–5 of the cycle (0.35 ± 0.03 ng/g wet tissue). Subsequently, it constantly increased to the highest value on days 14–18 (16.58 ± 0.52 ng/g wet tissue) and on days 19–21 it declined (11.10 ± 0.52 ng/g wet tissue). Progesterone at a low dose (10^–9 M ) resulted in significant (p < 0.05) increases and decreases in β‐END‐LI secretion by luteal cells from days 11–13 and 14–18, respectively. Higher doses of progesterone (10^–7 and 10^–5 M ) had no effect on β‐END‐LI release, compared with the control group. All dose‐levels of oxytocin used decreased β‐END‐LI secretion by luteal cells on days 11–13 and 14–18 of the cycle. Prolactin at doses of 0.1 and 1 ng/ml on days 11–13, and all doses tested on days 14–18 resulted in decreases in β‐END‐LI release from luteal cells. These results document evident changes in β‐END‐LI content in the pig corpus luteum during its development and indicate the potential roles of progesterone, oxytocin, and prolactin in luteal cell secretion of β‐END‐LI.     

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.     

Persistence and mobility of chlorsulfuron and metsulfuron under different soil and climatic conditions

The persistence and mobility of chlorsulfuron and metsulfuron were studied in Italy at four field locations. Chlorsulfuron was applied pre-emer-gence to wheat at 15 and 30 g a.i. ha^?1 in the autumn of 1986 and metsulfuron early post-emergence at 8 g a.i. ha^?1 in the spring of 1987. Soil samples were taken periodically at each location for 1 year. Trials were conducted until 1989 at three locations. After the wheat harvest in 1989, chlorsulfuron and metsulfuron residues in the soil were measured by bioassay in plots that had been treated annually for 3 years and also in plots that had received single treatments. Chlorsulfuron at both application rates dissipated to levels below 0.3 g a.i. ha^?1 over 12 months at three locations while, at the fourth, which received only 250 mm rain during the study, significant levels of residues were still present after 15 months. The degradation of chlorsulfuron applied at 30 g a.i. ha^?1 followed first-order kinetics, with half-lives of 149, 70, 59 and 51 days at the four locations. Chlorsulfuron mobility was high at three locations while, in the silty clay soil of the fourth, the herbicide was confined to the upper 30 cm, despite the high rainfall. Metsulfuron applied in spring was detected only in the first soil sample collection. It was present in all sampled layers at one location but, in the others, was confined to the surface layer. There was no evidence of herbicide accumulation in the soil with repeated applications. Persistance et mobilité du chlorsulfuron et du met-sulfuron dans différents sols et sous différentes conditions climatiques La persistance et la mobilité du chorsulfuron et du metsulfuron au champ ont étéétudiées en Italie sur quatre sites. Le chlorsulfuron était appliqué en pré-levée sur le blé aux doses de matière active (m.a.) 15 et 30 g ha^?1à l'automne 1986 et le metsulfuron en post-levée précoce à la dose 8 g m.a. ha^?1 au printemps 1987. Des échantillons de sol étaient prélevés périodiquement sur chaque site pendant 1 an. Les essais ont été conduits jusqu'en 1989 sur trois des sites. En 1989, après la récolte du blé. les résidus de chlorsulfuron et de metsulfuron dans le sol ont été mesurés par des essais biologiques dans des parcelles qui avaient été traitées chaque année pendant 3 ans, ainsi que dans des parcelles qui n'avaient reçu qu'un seul traitement. Le chlorsulfuron aux deux doses disparaissait jusqu'à des niveaux inférieurs à 0.3 g m.a. ha^?1 en 12 mois sur trois des sites, alors que sur le quatrième, qui n'a reçu que 250 mm de précipitations pendant l'étude, des résidus étaient présents en quantité significative après 15 mois. La dégradation du chlorsulfuron appliquéà la dose 30 g m.a. ha^?1 suivait une cinétique du premier ordre, avec des demi-vies 149, 70, 59 et 51 j sur les quatre sites. La mobilité du chlorsulfuron était élevée sur trois des sites alors que sur le quatrième oú le sol était une argile limoneuse, l'herbicide était confiné aux 30 cm superficiels en dépit de précipitations élevées. Le metsulfuron appliqué au printemps n'était détecté que dans le premier prélèvement. Sur un des sites, il était présent dans toutes les couches, mais était confinéà la couche superficielle sur les autres sites. Après des applications répétées, aucune accumulation d'herbicide n'était observée dans le sol. Persistenz und Mobilität von Chlorsulfuron und Metsulfuron in verschiedenen Böden und bei unterschiedlicher Witterung Die Persistenz und Mobilität von Chlorsulfuron und Metsulfuron wurden in Italien an 4 Standorten unter Freilandbedingungen untersucht. Chlorsulfuron wurde im Herbst 1986 bei Weizen im Vorauflauf mit 15 und 30 g AS ha^?1 und Metsulfuron im Frühjahr 1987 im frühen Nachauflauf mit 8 g AS ha^?1 angewandt. Von jeder Versuchsfläche wurden über einen Zeitraum von 1 Jahr periodisch Bodenproben entnommen. Die Versuche wurden an 3 Standorten bis 1989 durchgeführt. Nach der Weizenernte 1989 wurden mit Biotests die Rückstände von Chlorsulfuron und Metsulfuron in den Böden mit jährlicher (über 3 Jahre) sowie mit einmaliger Behandlung bestimmt. Die Chlorsulfuron-Rückstände nahmen an 3 Standorten bei beiden Aufwandmengen nach 12 Monaten auf 0,3 g AS ha^?1 ab. Am 4. Standort, wo während der Untersuchung nur 250 mm Niederschlag gefallen waren, wurden nach 15 Monaten noch signifikante Rückstände gefunden. Der Abbau von Chlorsulfuron folgte an allen Standorten bei einer Aufwandmenge von 30 g AS ha^?1 einer Kinetik erster Ordnung mit Halbwertszeiten von 149, 70, 59 und 51 Tagen. Die Mobilität von Chlorsulfuron war an 3 Standorten hoch, während es in dem schluffigen Tonboden des 4. Standortes trotz hohen Niederschlags in den oberen 30 cm verblieb. Das im Frühjahr ausgebrachte Metsulfuron wurde nur in der 1. Bodenprobenserie gefunden. An einem Standort lagen in allen beprobten Bodenschichten Rückstände vor, doch sonst waren sie auf die oberste Bodenschicht beschränkt. Bei wiederholter Anwendung konnte keine Herbizidakkumulation im Boden festgestellt werden.