Report on the analyses of mAb reactive with porcine CD8 for the second international swine CD workshop

Based on an analysis of their reactivity with porcine peripheral blood lymphocytes (PBL), only three of the 57 mAbs assigned to the T cell/activation marker group were grouped into cluster T9 along with the two wCD8 workshop standard mAbs 76-2-11 (CD8a) and 11/295/33 (CD8b). Their placement was verified through the use of two-color cytofluorometry which established that all three mAbs (STH101, #090; UCP1H12-2, #139; and PG164A, #051) bind exclusively to CD8⁺ cells. Moreover, like the CD8 standard mAbs, these three mAbs reacted with two proteins with a MW of 33 and 35 kDa from lymphocyte lysates and were, thus, given the wCD8 designation. Because the mAb STH101 inhibited the binding of mAb 76-2-11 but not of 11/295/33, it was given the wCD8a designation. The reactivity of the other two new mAbs in the T9 cluster with the various subsets of CD8⁺ lymphocytes were distinct from that of the other members in this cluster including the standards. Although the characteristic porcine CD8 staining pattern consisting of CD8^low and CD8^high cells was obtained with the mAb UCP1H12-2, a wider gap between the fluorescence intensity of the CD8^low and CD8^high lymphocytes was observed. In contrast, the mAb PG164A, not only exclusively reacted with CD4⁻/CD8^high lymphocytes, but it also failed to recognize CD4/CD8 double positive lymphocytes. It was concluded that this mAb is specific for a previously unrecognized CD8 epitope, and was, thus, given the wCD8c designation. A very similar reactivity pattern to that of PG164A was observed for two other mAbs (STH106, #094; and SwNL554.1, #009). Although these two mAbs were not originally positioned in the T cell subgroup because of their reactivity and their ability to inhibit the binding of PG164A, they were given the wCD8c designation. Overall, five new wCD8 mAbs were identified. Although the molecular basis for the differences in PBL recognition by these mAbs is not yet understood, they will be important in defining the role of CD8⁺ lymphocyte subsets in health and disease.     

Effect of soil moisture during stratification on dormancy release in seeds of five common weed species

Although the effects of cold stratification on the release of physiological dormancy in seeds have been studied extensively, knowledge of the role of soil moisture content on seed dormancy release during cold stratification is limited. Our study determined seed dormancy characteristics and the effect of soil moisture content on seed dormancy breakage during cold stratification in the five common weed species Amaranthus retroflexus, Chenopodium album, Chenopodium hybridum, Plantago lanceolata and Setaria glauca. Seeds of all five species were dormant at the time of harvest and their germination response to light and temperature varied. Soil moisture content had a significant effect on seed dormancy release of all species except P. lanceolata. Germination percentage of A. retroflexus, C. album, C. hybridum increased and then decreased as soil moisture content increased, regardless of germination test temperature. The optimal soil moisture content and seed moisture content for dormancy breakage of A. retroflexus, C. album, C. hybridum were 8%, 12%, 8% and 22.0%, 37.7%, 25.7% respectively. Dry storage (after‐ripening) significantly increased germination of S. glauca. Moreover, increasing soil moisture content first slowed and then increased dormancy breakage in S. glauca. These results suggest that data on soil moisture content should be incorporated into models that predict weed seed dormancy breakage and timing of seedling emergence as well as those for weed management.     

Escherichia coli O115 forms fewer attaching and effacing lesions in the ovine colon in the presence of E. coli O157:H7

Escherichia coli O115 has been isolated from healthy sheep and was shown to be associated with attaching-effacing (AE) lesions in the large intestine. Following previous observations of interactions between E. coli O157 and O26, the aim of the present study was to assess what influence an O115 AE E. coli (AEEC) would have on E. coli O157 colonisation in vitro and in vivo. We report that E. coli O115- and O157-associated AE lesions were observed on HEp-2 cells and on the mucosa of ligated ovine spiral colon. In single strain inoculum, E. coli O115 associated intimately with HEp-2 cells and the spiral colon in greater numbers than E. coli O157:H7. However, in mixed inoculum studies, the number of E. coli O115 AE lesions was significantly reduced suggesting negative interference by E. coli O157. Use of the ligated colon model in the present work has allowed in vitro observations to be extended and confirmed whilst using a minimum of experimental animals. The findings support a hypothesis that some AEEC can inhibit adhesion of other AEEC in vivo. The mechanisms involved may prove to be of utility in the control of AE pathovars.     

Paraneoplastic thrombocytosis-induced systemic thromboembolism in a cat

A six-year-old cat presented with clinical signs consistent with distal aortic thromboembolism while clinical signs of cardiovascular disease were absent. Diagnostics, including thoracic radiographs, electrocardiography, and echocardiography revealed no cardiovascular anomalies. Thoracic radiographs revealed multifocal pulmonary lesions consistent with neoplasia. Complete blood cell count demonstrated a marked thrombocytosis, leukopenia, and neutropenia. Histopathology of the pulmonary lesions confirmed multiple bronchoalveolar carcinomas. Myelodysplasia with megakaryocytic hyperplasia and ineffective myelopoiesis was noted on bone-marrow histopathology from multiple sites. The absence of other causes suggested a paraneoplastic thrombocytosis. The diagnosis of paraneoplastic thrombocytosis-induced thromboembolism was made due to the lack of underlying cardiac disease and the presence of a marked thrombocytosis. The presence of thrombocytosis and thromboembolism associated with neoplasia is discussed.     

SNOMED representation of explanatory knowledge in veterinary clinical pathology

BACKGROUND: The Systematized Nomenclature of Medicine (SNOMED) is an established standard nomenclature for the expression of human and veterinary medical concepts. Nomenclature standards ease sharing of medical information, create common points of understanding, and improve data aggregation and analysis. OBJECTIVES: The objective of this study was to determine whether SNOMED adequately represented concepts relevant to veterinary clinical pathology. METHODS: Concepts were isolated from 3 different types of clinical pathology documents: 1) a textbook (Textbook), 2) the Results sections of industry pathology reports (Findings), and Discussion sections from industry pathology reports (Discussion). Concepts were matched (mapped) by 2 reviewers to semantically-equivalent SNOMED concepts. A quality score of 3 (good match), 2 (problem match), or 1 (no match) was recorded along with the SNOMED hierarchical location of each mapped concept. Results were analyzed using Cohen's Kappa statistic to assess reviewer agreement and chi-square tests to evaluate association between document type and quality score. RESULTS: The percentage of good matches was 48.3% for the Textbook, 45.4% for Findings, and 47.5% for Discussion documents, with no significant difference among documents. Of remaining concepts, 40% were partially expressed by SNOMED and 14% did not match. Mean reviewer agreement on quality score assignments was 76.8%. CONCLUSIONS: Although SNOMED representation of veterinary clinical pathology content was limited, missing and problem concepts were confined to a relatively small area of terminology. This limitation should be addressed in revisions of SNOMED to optimize SNOMED for veterinary clinical pathology applications.     

Electroencephalographic observation on sheep and cattle with experimental cerebrocortical necrosis

Nine Suffolk sheep and 4 Holstein cattle were employed to observe electroencephalographic changes in experimental cerebrocortical necrosis. Amprolium (600 mg/kg/day) alone was given intraruminally 6 sheep and 4 cattle, all of which showed neurological signs and abnormal electroencephalograms. Both amprolium and thiamine (200 mg/day) and thiamine (50 mg/day) alone were given 2 and 1 sheep, respectively, as controls, which did not show any abnormal signs. Abnormal electroencephalograms included continuous slow waves and long-lasting spindles, both of which appeared diffusely and were seen subclinically or with neurological signs. The spindles consisted of slow waves or sharp- (or spike-) and-slow-wave complexes with or without convulsive seizures even during the convulsion stage. Conversely, the seizures occurred with the spindles or slow waves.     

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.