Effect of endocytic and metabolic inhibitors on the internalization and intracellular growth of Brucella abortus in Vero cells.

Uptake, transfer to rough endoplasmic reticulum, and intracellular growth of Brucella abortus were studied in Vero cells treated with endocytic and metabolic inhibitors. Infection of Vero cells was suppressed when inhibitors of energy metabolism (iodoacetate, dinitrophenol), receptor-mediated endocytosis (monodansylcadaverine, amantadine, methylamine), or endosomal acidification (chloroquine, ammonium chloride, monensin) were added to the inoculum. Inhibition was not observed when these drugs were added after the inoculation period. Infection of Vero cells by B abortus was inhibited by dibutyryl-cyclic adenosine monophosphate and Vibrio cholerae enterotoxin, but was stimulated by dibutyryl-cyclic guanosine monophosphate and escherichia coli heat-stable enterotoxin a. Uptake of B abortus by Vero cells was not prevented by colchicine, but was abolished by cytochalasin B. Uptake of heat-killed B abortus and noninvasive E coli was similar to that of viable brucellae. Intracellular growth of B abortus was not affected by cycloheximide. Results indicate that: B abortus may be internalized by a receptor-mediated phagocytic process; transfer of B abortus from phagosomes to rough endoplasmic reticulum may require endosomal acidification; and replication of B abortus within the rough endoplasmic reticulum may not depend on protein synthesis by the host cell.     

The cardiorespiratory effects of morphine and butorphanol in horses anaesthetised under clinical conditions

Twenty-five horses admitted for minor orthopaedic or soft tissue surgery were anaesthetised with detomidine, ketamine and halothane. Heart rate, arterial blood pressure, respiratory rate, tidal volume, minute volume, blood gases and occlusion pressures were measured before and for 30 mins after intravenous (iv) injection of saline, butorphanol 0.05 mg/kg bodyweight (bwt) or morphine 0.02 or 0.05 mg/kg bwt. Drug or saline treatment induced no significant changes from pre-treatment values within a group for arterial blood pressure, heart rate, respiratory rate, arterial carbon dioxide tension, arterial oxygen tension and occlusion pressure. In conclusion, both morphine and butorphanol at the stated doses cause no adverse effects on the cardiovascular and respiratory systems of anaesthetised horses.     

Confirmation of rinderpest in experimentally and naturally infected cattle using microtitre techniques

Summary Virulent rinderpest virus was detected by immunoperoxidase staining of microtitre bovine kidney cell cultures within 24 to 48 hours of inoculation with prescapular lymph node and spleen homogenates from experimentally infected steers. Rinderpest virus specific cytopathic effects were evident from 48 hours in microtitre plates and from 72 hours in rolled tube cultures. Nasal and ocular secretions collected from cattle naturally infected with rinderpest and inoculated into bovine kidney cell cultures did not readily yield cytopathic virus in both tubes and microtitre plates, but immunoperoxidase staining of microtitre cultures on the fourth day of inoculation detected replication of virus in cultures inoculated with ocular and nasal secretions from seven of 17 cattle tested.

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Resumen Se detectó el virus virulente de mediante la tinción con inmunoperoxidasa de cultivos de células de ri?ón bovino en bandejas de microtitulación, después de la inoculación de estos con suspensiones homogenizadas de ganglios linfáticos preescapulares y de bazo provenientes de novillos infectados experimentalmente. El efecto citopático del virus de la peste bovina fue evidente desde las 48 horas en bandejas de microtitulación y desde las 72 horas en tubos de cultivo giratorios. Secreciones oculares y nasales colectadas de ganado infectado en forma natural con la peste bovina e inoculadas en cultivos de células de ri?ón bovino, no mostraron efecto citopático fácilmente en tubos giratorios o bandejas de microtitulación, pero la tinción de las bandejas con inmunoperoxidasa reveló replicación del virus a partir del cuarto día de inoculación con secreciones oculares y nasales en siete de los 17 animales examinados.

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Résumé Un virus bovipestique virulent a été décelé par le test de coloration à l'immunoperoxydase de cellules rénales bovines en culture dans des plaques de microtitrage et infectées 48 heures plus t?t avec des homogénats de ganglions lymphatiques et de rate provenant de bouvillons infectés expérimentalement. Les effets cytopathogènes du virus étaient évidents au bout de 48 h dans les plaques de microtitrage et 72 h dans les tubes en rollers. Les sécrétions nasales et oculaires prélevées sur du bétail infecté naturellement par la peste bovine et inoculées sur des cellules rénales bovines n'ont pas toujours montré d'effet cythopathogène aussi bien dans les tubes que dans les plaques de microtitrage. Cependant, la coloration à la peroxydase au jour 4 après l'inoculation a permis de déceler la présence de virus dans 7 cas sur 17.

     

Anaerobes in periodontal disease in the dog: a review

As anaerobic sampling and culture techniques improved, the documented prevalence of anaerobic bacteria in periodontal disease has increased. The anaerobic bacteria have become more well-known in humans and consequently in dogs, since this species is a major model in periodontal studies. A review of the literature related to anaerobic flora is described.     

Salmonella contamination of poultry flocks in The Netherlands

The contamination of poultry in the Netherlands with Salmonella enteritidis was tested. For this, different methods (detection of S. enteritidis in faecal samples of 25 g; detection of S. enteritidis in cloacal swabs; detection of S. enteritidis by serological testing of antibodies in serum) were compared for their efficiency to detect S. enteritidis in flocks of poultry. Testing of faecal samples clearly yielded the best results. This method was used in a transmission study, in which 14 flocks descending from a contaminated primary mother flock were screened for the presence of S. enteritidis. The method was also used for screening 49 flocks of laying hens and 52 flocks of broiler chickens throughout the Netherlands. From the transmission study it became clear that S. enteritidis, phage type 2 (Dutch phage set) was isolated both from the mother flock and from five of the descendent flocks. Screening of poultry flocks for the presence of salmonella revealed that salmonella was present in 47% of the layer flocks and in 94% of the broiler flocks. S. enteritidis was isolated from 15% of the flocks screened.     

Frustrating case presentations in canine theriogenology.

The practice of small animal theriogenology is rewarding, but frustrations exist concerning technologic advances as compared with other species. Reproductive clinicians striving to practice good quality medicine readily identify topics of common concern: causes that are not identified or therapeutics that are not available or applicable. Improved collaboration among theriogenologists specializing in small animal practice is evidenced by growing attendance at national and international scientific meetings, increased scientific publications, and internet communications.     

Isolation and purification of a diagnostic antigen for bovine cysticercosis by hydrophobic chromatography

An ammonium sulfate fraction of Taenia hydatigena cyst fluid (ThFAS) was further fractionated by hydrophobic interaction chromatography, using alkylagarose and omega-amino alkylagarose columns, in an effort to isolate and purify a specific diagnostic antigen in the ThFAS preparation. The less than 12 kDa antigen was found to have an affinity for immobilized alkanes with chain length of six carbons or greater. The antigen was recovered in an ethylene glycol eluate from a hexylagarose column then analyzed by Western blot; it reacted with bovine and human cysticercosis infection sera and with specific monoclonal antibodies but not with control sera or fascioliasis infection sera. When the eluate was used as coating antigen in a plate ELISA assay no false positive reactions were seen in sera from cattle infected with Fasciola hepatica; false positive reactions were observed for the unfractionated ThFAS antigen preparation.