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.     

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.     

Isotypic antibody responses in cattle infected with Haemophilus somnus

Bovine antibody responses to Haemophilus somnus were compared on the basis of clinical and bacteriological findings. Serum IgG1 and IgM antibody titres were significantly increased in clinically normal cattle that were bacteriologically positive for H somnus from the nasal or vaginal mucosae compared with clinically normal, negative cows. IgG2 titres did not differ significantly between these two groups. However, IgG2 antibody was significantly higher in animals with H somnus disease (pneumonia or abortion) than in clinically normal cattle (whether bacteriologically positive or negative), while IgG1 and IgM titres did not differ between diseased and bacteriologically positive, clinically normal cattle. These antibody trends were duplicated in experimental H somnus abortion or pneumonia, with the greatest response occurring within the IgG2 subclass. Cattle vaccinated systemically with killed whole H somnus produced a predominant IgG2 response with minimal IgG1 and IgM responses. These results demonstrate that IgG2 antibody is consistently elevated in H somnus disease, and suggest that this response may be useful in discriminating diseased from asymptomatic cattle.     

Electromyography and spinal evoked potentials in cauda equina syndrome of dogs

In order to investigate usefulness of electrodiagnostic methods in Cauda Equina-Compression, 26 dogs with this disease were subjected to electromyography and spinal evoked potentials. 16 normal dogs served as controls. It was found that--after stimulation of the Nn.tibialis, peronaeus, pudendus and coccygeales and recording at the lumbosacral junction--latencies and nerve-conduction velocities were largely normal. However the amplitudes were decreased and--depending on the severity of the compression--the morphology of the potentials was altered although considerable standard deviations were obtained. Electromyographically, increased spontaneous activity with fibrillations and positive sharp waves were found in the myotomes of the lumbosacral nerves. Their number increased according to the severity of the cauda equina compression and they had a centrifugal distribution pattern. Based on these results it appears indicated to perform an electromyographic exam before applying myelography. The technique of spinal evoked potentials will need more refinement and standardisation before being useful in a clinical situation.     

Biomicroscopy of the tear film: the tear film of the pekingese dog

Polarised light biomicroscopy was used to examine the normal pre-corneal tear film in 21 eyes of 12 pekingese dogs. The purpose of the study was to examine the influence of excessive exophthalmos on the pre-corneal tear film in the dog. The majority of the animals were found to have high levels of ocular surface contamination by particulate material and plaques of viscous mucus. Other abnormalities included surface lipid with an abnormal granular (three dogs) or 'curdled' (two dogs) appearance; excessive thinning of the lipid layer of the tear film; and the presence of dark globular structures in two dogs, which were presumed to be abnormal meibomian lipid. Break up of the tear film was observed in one dog. Grossly, a thread of viscous mucus was frequently observed along the margin of the lower eyelid. It is postulated that this thread forms because of the excessively exophthalmic conformation of the breed, which prevents the normal access of effete mucus and entrapped debris to the lower conjunctival fornix. The combination of the above factors in the pekingese is suggested as the mechanism whereby the tear film has a reduced stability, thus enhancing the risk from factors more usually considered to initiate corneal ulceration in the breed. The possible adverse effects of lid splitting for the mass removal of distichiae in exophthalmic dogs is discussed.     

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.