The protein patterns of sodium dodecyl sulfate-solubilized whole cells of porcine strains of Haemophilus were studied by using polyacrylamide gel electrophoresis. The pattern of Haemophilus pleuropneumoniae was very homogenous and was independent of the serological type. At least two different patterns could be distinguished in Haemophilus parasuis, suggesting some heterogeneity in this species. The results were highly reproducible and were not affected by growth conditions. Comparable patterns were obtained after solubilization with sodium taurocholate and sodium carbonate and after ultrasonic treatment, but not after phenol-acetic acid extraction. In addition, we compared porcine strains with human strains (Haemophilus influenzae, Haemophilus parahaemolyticus) and also with some bacteria of the genera Pasteurella, Actinobacillus, Brucella, Moraxella, and Bordetella. The species-specific picture is given mostly by the pattern of a group of proteins with molecular weights just above 68,OOO (Haemophilus) and in the region of molecular weights between 23,000 and 40,000 and between 15,000 and 17,000 (Haemophilus, Pasteurella, Actinobacillus) . These observations suggest the possible use of this method as an aid in studying the taxonomy of these bacteria.The electrophoretic patterns obtained by polyacrylamide gel electrophoresis (PAGE) of cell proteins from bacteria has proved to be useful in taxonomic studies. Since these cell proteins are genetically directed, their patterns tend to express genetic relationships between microorganisms (29).In recent years, reports have been published on the use of PAGE with a great variety of microorganisms, ranging from mycoplasmas (5, 6, 27-29,40) to bacteria and even fungi (Aspergillus fumigatus) (38); the bacteria have included brucellae (20), staphylococci (3), enteric bacteria (26, 30, 32, 33, 44), streptomycetes (7), yersiniae (lo), mycobacteria (8), streptococci (17,18), campylobacters (vibrios) (21), neisseriae (24, 31, 45), Corynebacterium diphtheriae (16), Erysipelothrix rhusiopathiae (43), and Pasteurella haemolytica (42). For the genus Haemophilus, only a few studies of this type have been reported. Neumann and Him (22) found that the protein patterns of strains of Haemophilus paragallinarum, Haemophilus avium, Haemophilus parainfluenzae, and Haemophilus parasuis are quite different, even among strains within the same species, so that the method has been judged to be unreliable for taxonomic purposes.It has been clearly demonstrated that the cytoplasmic membranes and whole-cell proteins of mycoplasmas and also of L-forms of bacteria show highly reproducible and species-specific electrophoretic patterns with PAGE (28,39,40).It is likely in such cases that PAGE can be used as a taxonomic tool. In the case of other bacteria, the majority of authors have observed similarities in the electrophoretic patterns within a genus (20, 32, 41) or within a species (7, 8, 10, 18, 20, 24, 31, 42) but not between genera (20, 32, 41).Despite the rather complex structure of bacterial ce...

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Several Mycoplasma and Acholeplasma species chosen at random and solubilized with sodium dodecyl sulphate showed a common periodic acid-Schiff positive band with an apparent molecular weight of about 64 000, when examined by polyacrylamide gel electrophoresis. Another more cathodic minor band was detected in M. hyopneumoniae and M. flocculare. The common periodic acid-Schiff positive band appeared when a precipitate of serum constituents of the uninoculated growth medium after incubation was examined. The minor band was identified as a serum glycoprotein contaminating mycoplasmas grown in the presence of swine serum. We draw attention to the compounds as a possible source of error in serological tests or in the lymphocyte stimulation response. After lithium diiodosalicylate solubilization and aqueous phenol extraction, polyacrylamide gel electrophoresis showed a periodic acid-Schiff positive band in membranes from M. hyopneumoniae (molecular weight 75 000) and M. hyorhinis (molecular weight 80 000), suggesting the presence of a membrane glycoprotein. Such a glycoprotein was absent from A. granularum. Since the common periodic acid-Schiff positive band was not extracted by aqueous phenol, this growth medium constituent did not contaminate the preparations of membrane glycoproteins. However, the minor band was present in glycoprotein preparations of M. hyopneumoniae grown in the presence of swine serum.

Zusammenfassung Mykoplasmen und Acholeplasmen von Rind, Schaf, Ziege und Schwein wurden elektrophoretisch in SDS Polyacrylamid‐Gel mit Hilfe der „Slab”‐Technik untersucht. Auf Grund der eindeutig verschiedenen Elektropherogramme all dieser Stämme wird diese Methode als routinediagnostischer Test empfohlen. Zu diesem Zweck wird der Test vereinfacht durch Vernachlässigung der Proteinmessung. Diese Methode wurde für die Diagnostik einer erkrankten Schafherde angewendet, wobei 47 M. ovipneumoniae und 3 M. arginini nachgewiesen worden sind. Diese Ergebnisse stimmten mit anderen Identifizierungsmethoden überein. Summary Use of polyacrylamide — gel — electrophoresis in diagnosis of farm animal mycoplasmas Mycoplasmas and Acholeplasmas from cattle, sheep, goats and pigs were studied by electrophoresis in SDS‐polyacrylamide‐gel with the help of the “slab” technique. On the basis of the clearly different electrophoretograms of all the species this method is recommended for routine diagnosis. To this end the technique is simplified by omitting the protein determination. The method was used for diagnosis in an affected sheep flock and 47 isolations of M. ovipneumoniae and 3 of M. arginini were made. The results agree with those of other methods of identification. Résumé Utilisation de l'électrophorèse sur gel de polyacrylamide dans le diagnostic des mycoplasmes chez l'animal de rente Des mycoplasmes et acholeplasmes du bovin, du mouton, de la chèvre et du porc ont été examinés électrophorétiquement dans du gel de polyacrylamide SDS à l'aide de la technique «Slab». Sur la base des électrophérogrammes nettement différents de toutes ces souches, on recommande cette méthode comme test de diagnostic pour la routine. Le test est simplifié dans ce but par l'abandon de la mesure des protéines. Cette méthode fut utilisée pour le diagnostic d'un troupeau de moutons malades dans lequel 47 M. ovipneumoniae et 3 M. arginini ont été mis en évidence. Ces résultats correspondaient avec d'autres méthodes d'identification. Resumen Uso de la electroforesis sobre gelosa de poliacrilamida en el diagnóstico de mycoplasmas en los animales domésticos Se examinaron electroforéticamente mycoplasmas y acoleplasmas de vacunos, oveja, cabra y cerdo sobre gelosa de poliacrilamida SDS con ayuda de la técnica „Slab”. En vista de los diversos electroferogramas inequívocos de todas estas estirpes, se recomienda el método presente como prueba diagnóstica rutinaria. Para este fin se simplifica la prubea omitiendo la medición proteica. Se recurrió a este método para establecer el diagnóstico en un rebaño de ovejas enfermas, poniendo en evidencia 47 M. ovipneumoniae y 3 M. arginini. Estos resultados coinciden con los obtenidos con otros métodos de identificación.