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.     

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.     

Nephrolithiasis resulting in intermittent ureteral obstruction in a cow

Bilateral nephrolithiasis with intermittent ureterolithiasis was diagnosed in a 7-year old Holstein cow. Two episodes of ureterolithiasis resulted in severe azotemia which resolved after spontaneous movement of the stone. A third episode of obstruction one year after the initial episode resulted in rupture of one kidney, necessitating euthanasia. The histopathological examination of the kidney was diagnostic for chronic pyelonephritis. Corynebacterium sp. was cultured from a nephrolith. In this case it is believed that the chronic pyelonephritis predisposed to the calculi formation.     

Effect of a new pelleting process on the level of contamination of poultry mash by Escherichia coli and Salmonella

The efficacy of a new pelleting process in eliminating naturally occurring Escherichia coli and salmonella from poultry mash was assessed by comparing the microbial load in raw and processed mash. Instead of using steam produced in a boiler, the new process conditioned mash in an Original Vertical Conditioner with steam and other hot gases generated by direct combustion in a Vaporator. E. coli was isolated from 72-100% of samples of raw mash in all trials, and the mean number of colony-forming units of E. coli/g of samples was 6.8 +/- 4.0 X 10(4). Salmonellae (S. senftenberg, S. bredeney, and S. mbandaka) were isolated from 5-10% of samples of raw mash utilized in three of the seven trials. Within limitations of the sampling and analytical tests utilized, the new pelleting process eliminated salmonella from all mash in which the organism was known to be present but eliminated E. coli in only three trials. The process appeared to be 100% effective against both organisms when mash entering the pellet mill from the conditioner had a temperature of 85.7 C and a moisture content of 14.5% and had been retained and treated with steam and hot gases for 4.1 minutes.     

Differentiation of hog cholera and bovine virus diarrhoea viruses in pigs using monoclonal antibodies

Monoclonal antibodies against hog cholera and bovine viral diarrhoea viruses were assayed on organ tissue sections of experimentally infected animals. The animals had been infected simultaneously with both viruses. The antibodies were tested using an indirect immunofluorescence test and an indirect enzyme immunoassay with a biotin/streptavidin/peroxidase detection system. A polyclonal hyperimmune serum was used as a control in direct immunofluorescence tests. Both techniques based on monoclonal antibodies were more sensitive and more specific than the conventional test, the enzyme immunoassay being more sensitive than the immunofluorescence test. Small amounts of BVD viral antigen were demonstrable with monoclonal antibodies in most organ tissues.     

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.     

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.