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.     

Use of autogenous cancellous bone graft for treatment of osteolytic defects in the phalanges of three cattle.

Osteolytic defects were detected radiographically in the distal sesamoid bone of a 16-month-old Bralers heifer, in the middle phalanx of a 14-month-old American Gray Brahman bull, and in the distal phalanx of a 3-year-old American Gray Brahman bull. The articular cartilage was damaged in each animal because of osteolysis or pathologic fracture. After each animal was anesthetized and positioned in lateral recumbency, the lesions were curetted and packed with cancellous bone harvested from the same animal's tuber coxae. Basic postoperative management involved stall rest and immobilization of the graft site with a fiberglass cast (42 to 79 days), after which a support bandage was used for approximately 2 weeks. Recurrence of lameness has not been observed in these animals for 60 months, 58 months, and 21 months, respectively. These cases exemplify the benefit of using an autogenous cancellous bone graft for treatment of severe osteolysis of a digit in cattle.     

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.     

Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to bovine virus diarrhoea virus in milk

The present study shows that milk is an appropriate source for detection of seroreactors to bovine virus diarrhoea virus (BVDV). There was close agreement between antibody titres in serum and in skim milk, as determined by an indirect enzyme-linked immunosorbent assay. The antibody titres were usually lower in skim milk than in serum, but all seropositive cows (n = 84) were also skim milk-positive and all but one seronegative cow (n = 55) proved negative in skim milk. During lactation, the level of antibodies to BVDV in milk showed an inverse relationship to the amount of milk produced. However, there was a sufficient level of antibodies in milk throughout lactation to permit an adequate determination of BVDV antibody status in dairy cows. There was a mutual good agreement between milk antibody titre in the four mammary quarters, irrespective of milk cell count. Milk can be used to detect seroreactors to BVDV. Milk is preferable to blood in large-scale epidemiological studies, since the sampling procedure is much simpler.     

Cytological, Immunocytochemical and Ultrastructural Study of the Adenohypophyseal Pars Distalis of the Kid (Capra hircus): The TSH cell

The structure and ultrastructure of the adenohypophyseal pars distalis in kids of the Murciano-granadiana breed (18 males and 12 females), aged 2-3 months and with an average carcass weight of 8 kg has been studied. Techniques of staining (Tetrachrome Herlant's, and Cleveland-Wolfe) and histochemistry (PAS, PAS-OG and BA-PAS-OG) contrasted with immunolabelling (PAP) have been used. In addition an ultrastructural study has been made and nucleus and cytoplasm, secretory granules, mitochondria and lysosomes have been measured with a semiautomatic image analyzer (IBAS-1). TSH cells are found in sagittal section in the anterior area and in an antero-caudodorsal band, and transversally in the ventral and medial region; they are 6% and their average size is 14.15 microns. Ultrastructurally they are a single cellular type with spherical granules whose size is 195 nm in males and 149 in females; these granules are characterized by a clear halo and a variable electronic density. The rough endoplasmic reticulum is found as slightly enlarged vesicles with a homogeneous and moderately electro-dense content or in concentric stratifications.     

The relationships of birth weight, preweaning gain and postweaning gain with the bovine major histocompatibility system

A total of 739 cattle from nine breeds maintained at the Roman L. Hruska U.S. Meat Animal Research Center, Clay Center, Nebraska were tested for 42 class I antigens of the bovine major histocompatibility system (BoLA). Each antigen appears to be the product of a distinct co-dominant allele of the BoLA-A locus. The number of antigens present in each breed ranged from a minimum of 10 in Hereford to a maximum of 21 in Charolais cattle. There were large differences among breeds in the frequencies of antigens. The effect of each antigen on birth weight, preweaning weight gain and postweaning weight gain was estimated in a gene substitution model. Each breed was analyzed separately. There were significant effects of some BoLA antigens on birth weight, preweaning weight gain and postweaning weight gain, which is consistent with previous reports showing associations between the major histocompatibility system and growth parameters in mice, rats and pigs. However, further research is necessary to confirm these findings and to determine the biological mechanisms underlying these associations.     

Evaluation of three strip tests for measuring electrolytes in mares' pre-partum mammary secretions and for predicting parturition

Three strip test kits which measure calcium and/or magnesium in mares' mammary secretions were compared and assessed for their accuracy in:- 1) measuring these electrolytes compared to standard laboratory measurements, and 2) predicting the time of parturition. Daily samples of mammary secretions were obtained pre-partum from 35 Thoroughbred mares and allocated a score. Measurement of mammary secretion calcium and/or magnesium by the strip tests and laboratory methods correlated significantly (P less than 0.001). The results obtained for prediction of parturition were analysed by two methods. Method i) indicated that the majority of mares had a maximum score on the day of birth. However, the second method of analysis included all prepartum scores and this demonstrated that maximum scores occurred on more occasions prior to the day of birth than on the day of birth itself. The number of nights spent attending mares, and the percentage of occasions when foaling was missed, were also calculated for given strip test scores. The results indicate that the test kits are not particularly accurate in predicting time of parturition although they are a reliable means of measuring mammary secretion calcium and/or magnesium. It is suggested that the test kits may be more helpful by indicating when it is not necessary to attend mares at night.     

Ultrastructure of sarcocysts from water buffalo in India

The ultrastructure of sarcocysts of macro- and microscopic species of Sarcocystis was compared from naturally infected water buffalo from India. Grossly visible sarcocysts had walls consisting of cauliflower-like villar protrusions, typical of S. fusiformis. The sarcocyst wall of the microscopic species of Sarcocystis was 6.4 microns thick and consisted of tightly packed conical villar protrusions that were 9.6 microns long and 3.7 microns wide at the base. At approximately 3 microns above the base, the distal two-thirds of the villar protrusion became conical shaped and was bent laterally at an angle of 45 degrees to the sarcocyst surface. The granular layer beneath the villar protrusions was 0.9 microns thick. In S. levinei the granular layer was 1.9 microns thick, the villar protrusions were narrow and it had a highly undulating primary cyst wall. Whether the microscopic S. levinei-like sarcocysts of Indian and Malaysian water buffalo are distinct species of Sarcocystis will require further investigation.     

Calcium fluxes across the wall of the ovine reticulorumen in vivo

Calcium fluxes were measured, in vivo, in both directions across the ovine reticulorumen wall of four sheep when the luminal potassium concentration was either 30 mmol litre-1 or 90 mmol litre-1. Neither fluxes were affected by an increased potassium concentration although net magnesium absorption was decreased (PO less than 0.05) and the transmural potential difference was increased (P less than 0.01) under these conditions. The results obtained suggest that, unlike magnesium, calcium is transported bidirectionally across ovine ruminal tissue independently of the transmural potential difference across the rumen wall.