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Phagocyte released reactive oxygen species are often discussed in connection with ischemic and reperfusion injuries to the myocardium. The kinetics of the accumulation and oxidative burst of human blood phagocytes was studied by chemiluminescence during open heart surgery in the myocardium of human patients. Direct evidence is presented for an accumulation of neutrophils along with their markedly increased metabolic activity (oxygen radical formation), especially following the reperfusion of the ischemic myocardium. Leukocyte numbers and activity remained significantly elevated even in the venous blood obtained 24 h after the operation.

SummaryThe influence was studied of changes of the bacterial host strain on the course of inactivation of Staphylococcus phages A/5 and ϕ131 after u.v.-irradiation, gamma-irradiation and photodynamic inactivation by AO. Effectiveness of phage plating was alike on all the host strains used. An influence of the various bacterial host strains on the survival of inactivated phages was found for u.v.-irradiated phages only. The bacterial host strains could be divided into strain groups having a favourable effect on the inactivation course of u.v.-irradiated phages, i.e. increasing the number of surviving phage particles, and into bacterial strains which may further reduce the survival of inactivated phage particles. The possible presence of repair mechanism in Staphylococcus bacterial cells is discussed.

SummaryPhage 55 of Staphylococcus aureus strain 55, which is extraordinarily sensitive to dark inactivation with acridine orange, loses this property after a multiple passage through its own host, until it is almost entirely resistant to the dark effect of acridine orange. Such stocks are also less sensitive to inactivation by other photodynamically-active dyes. However, the morphological properties of the phage 55, as well as its host range spectrum of activity, do not change. However, it is probable that some further properties of phage 55 changed in the course of passages, a matter of considerable importance in a phage of an international series.

The production of autoplaques in clinical strains of Pseudomonas aeruginosa is very frequent andcauses considerable difficulties in work with these cultures, especially during their evaluation. There are optimum media for the demonstration of this type of autolysis. We have succeeded in the isolation of stable AP+ and AP- cultures. Under the effect of passages and different media, the majority of cultures changed the irability of autolysis. Hospitals, waste-baskets, sinks, etc. represent reservoirs of AP+ strains of P. aeruginosa. In older cultures, cells surviving within the autoplaques may be isolated more easily than those from a culture of the same age but without autolysis. The protective effect of autolysis on some P. aeruginosa cells cannot be excluded.

SummaryThe action of the photodynamically active radiomimetic acridine orange on several phages of Staphylococcus of known morphology, serological type, phage type and the base composition in DNA has been studied. For these phages, the inactivation rate has been investigated, both at photodynamic inactivation with 10 μg/ml. AO and at inactivation in the dark with a tenfold concentration of the dye. Taxonomically related bacteriophages were inactivated with AO to approximately the same extent, whereas no correlation has been found between the rate of inactivation in the dark and the serological groups of Staphylococcus phages.

Summary: The present report gives the results of the effect of 10 bacteriophages of Serratia marcescens on 107 strains of the genus Serratia. The phage S103 was the most effective (80% strains lyzed), and the phage PK the least (12% strains lyzed). Only 13 strains were resistant to all the phages studied. The majority of the strains of the genus studied could be allocated to one of seven groups on the basis of their sensitivity to the phages.

Pseudomonas-aeruginosa-Phagen wurden durch UV-, X-Strahlen und Afridinorange inaktiviert und die erzielte Wirkung auf die Differenzierung der DNS oder RNS enthaltenden Bacteriophagen untersucht. Es ergaben sich eine auffallend schwache Inaktivierung durch Akridinorange im Dunkelversuch und gegen Strahlenwirkung ausgesprochen sensible Phagen mit einfaseriger DNS und RNS.

Fifty-six phages designated the U-series and 34 phages designated the Ph-series were carefully examined. The spectrum of activity was established for 183 strains of Staphylococcus epidermidis from different countries and for 258 strains of Staphylococcus aureus. All 90 phages were active against strains of both species, but some differences in sensitivity were noted. Sixteen of the 90 phages were selected for phage typing of S. epidermidis. With use of a routine test dilution of phage (the highest dilution that yields confluent lysis of the propagation strain), 71.6% of 183 coagulase-negative staphylococci tested were sensitive to the phages; 42 phage patterns were observed. The results indicate that phage typing of S. epidermidis can be recommended for epidemiological and ecological purposes.

A modification of the "one-plate method" for detection of staphylococcal hemolysins has been described. Bacterial strains of Staphylococcus aureus, Streptococcus agalactiae and Cornebacterium pseudotuberculosis were replaced by their prepurified hemolytically active exosubstances: beta-toxin, CAMP-factor, and, the exosubstance of Corynebacterium pseudotuberculosis called COREX. By means of the modification described, exact detection of hemolysins in possible with staphylococcal strains producing one type of hemolysin as well as with strains producing a combination of hemolysins.

ummary In blood agars containing erythrocytes from ruminants, hemolysis due to Streptococcus agalactiae, and streptococci of group B, is enhanced by the hemolytically active exosubstance of Corynebacterium pseudotuberculosis. The effect is displayed by both native and prepurified exosubstance. Unlike staphylococcal beta-toxin, corynebacterial exosubstance does not enhance hemolytic activity of Streptococcus uberis strains, not even of the CAMP-positive ones. The corynebacterial exosubstance, denoted by the authors as COREX, is of use for rapid identifcation of Streptococcus agalactiae and its differentiation from Streptococcus uberis. It is usable in tests with pure cultures as well as added to the substrate for identification of Streptococcus agalactiae in primary culture. Zusammenfassung Die Verwendbarkeit der hamolytisch aktiven “Exosubstanz” von Corynebacterium pseudotuberculosis bei der Isolierung und Bestimmung von Streptococcus agalactiae und ihr Vergleich mit dem beta-Toxin von Staphylococcus aureus Auf Blutagar-Nahrboden, die Erythrozyten von Wiederkauern enthalten, wird die von Streptococcus agalactiae, bzw. von Streptokokken der Gruppe B verursachte Hamolyse durch die hamolytisch aktive “Exosubstanz” von Corynebacterium pseudotuberculosis potenziert. Diese Fahigkeit besitzt die “Exosubstanz” in nativer und in vorgereinigter Form. Zum Unterschied vom Staphylokokken-Beta-Toxin verstarkt die “Exosubstanz” nicht die hamolytische Aktivitat der Streptococcus uberis-Stamme, auch nicht die der CAMP-positiven. Die “Exosubstanz”, von den Verfassern als COREX bezeichnet, ist fur die schnelle Diagnostik von Streptococcus agalactiae, vor allem im Hinblick auf seine Unterscheidung von Streptococcus uberis, geeignet. COREX ist auch als Zusatz zum Nahrboden, fur die Bestimmung von Streptococcus agalactiae in Primokulturen, verwendbar. Resume Valeur diagnostique de l'exosubstance hemolytique du Corynebacterium pseudotuberculosis pour l'isolation et l'identification du Streptococcus agalactiae et sa comparaison avec la beta-toxine du Staphylococccus aureus Au agar du sang, contenant des erythrocytes des ruminants, l'hemolyse produit par Streptococcus agalactiae et par streptococci de la groupe B est renforcee par l'activite hemolytique de l'exosubstance du Corynebacterium pseudotuberculosis. Cet effet est exerce egalement par l'exosubstance corynebacterielle native et prepurifie. Contrairement a la beta-toxine du Staphylococcus, elle ne renforce pas l'activite hemolytique des souches du Streptococcus uberis, meme de celles qui sont CAMP-positives. L'exosubstance corynebacterielle, designee par les auteurs comme COREX, est apte a la diagnose rapide du Streptococcus agalactiae et son differencement du Streptococcus uberis. COREX est applicable en essais avec cultures pures et aussi pour supplementer le agar a determiner Streptococcus agalactiae en culture primaire. Resumen Valor diagnostico de la exosubstancia hemoliticamente activa producida por Corynebacterium pseudotuberculosis y comparacion de esta con la beta-toxina de Staphylococcus aureus en el aislamiento e identificacion de Streptococcus agalactiae Una exosubstancia hemoliticamente activa producida por Corynebacterium pseudotuberculosis aumenta el efecto hemolitico de Streptococcus agalactiae y otros estreptococos del grupo B en agares-sangre con eritrocitos de rumiantes. El fenomeno se observa usando bien la forma natural, bien la forma prepurificada de dicha exosubstancia. A diferencia de la toxina beta estafilococica no aumenta hemolisis de las cepas de Streptococcus uberis que dan reaccion positiva a la prueba CAMP. La exosubstancia corinebacteriana, designada COREX por los autores de este articulo, se puede emplear para el diagnostico de Streptococcus agalactiae y su diferenciacion de Streptococcus uberis tanto en una prueba con cultivos puros, como agregandola al medio selectivo, para la identificacion de Streptococcus agalactiae en cultivos primarios de muestras investigadas.

Selected 22 phages of coagulase-negative staphylococci were tested against group-specific antisera for Staphylococcus aureus phages using a neutralization test. Fourteen of the tested phages were classified into the serological group B and one into the group F. All other phages were resistant to the antisera against S. aureus phages but cross-reacted with the antisera produced against themselves. Therefore they could be joined together into a new serological group of the staphylococcal phages.

Summary Haemolytically latent active exosubstances from Streptococcus agalactiae (the so called CAMP-factor) as well as from Streptococcus uberis (called in our report UBERIS-factor) can be prepared in a prepurified form by precipitation with acetone. Both exosubstances produced complete haemolysis of ovine erythrocytes in the zone of staphylococcal beta-toxin. In the zone of the exotoxin from Corynebacterium pseudotuberculosis complete haemolysis was seen with CAMP-factor only. The phenomenon described in the zone of beta-toxin with both CAMP-factor and UBERIS-factor is inhibited by factor(s) present in foetal, normal and hyperimmune sera. CAMP-factor and UBERIS-factor display immunogenic properties and react with homologous antisera in the double-gel-immunodiffusion test. The two exosubstances show different antigenic properties, and both of them differ from staphylococcal delta-toxin. Consequently, they could offer a means of supplementary serotyping of Streptococcus agalactiae and CAMP-positive strains of Streptococcus uberis. Zusammenfassung Vergleich einiger Eigenschaften des CAMP-Faktors von Streptococcus agalactiae mit der latent hamolytisch aktiven Exosubstanz von Streptococcus uberis Durch Prazipitierung mit Azeton konnen latent hamolytische Exosubstanzen in vorgereinigter Form sowohl von Streptococcus agalactiae (den sog. CAMP-Faktor), als auch von Streptococcus uberis (von uns UBERIS-Faktor bezeichnet) gewonnen werden. In der Zone des Staphylokokken-Betatoxins bewirken beide Exosubstanzen eine vollstandige Hamolyse der Schaferythrozyten; in der Zone des Exotoxins aus Corynebacterium pseudotuberculosis wird eine derartige Hamolyse nur beim CAMP-Faktor beobachtet. Das fur CAMP- und UBERIS-Faktor in der Betatoxin-Zone beschriebene Phenomen wird gehemmt durch Faktor(en), die in fotalen, normalen, sowie hyperimmunen Blutseren anwesend sind. Der CAMP- und UBERIS-Faktor sind immunogen und im “double-gel-diffusion”-Test reagieren sie mit homologen Antisera spezifisch. Beide Exosubstanzen besitzen unterschiedliche antigene Eigenschaften und beide unterscheiden sich auch vom Staphylokokken-delta-Toxin. Dadurch bietet sich die Moglichkeit einer zusatzlichen Serotypisierung von Streptococcus agalactiae und CAMP-positiver Streptococcus uberis-Stamme. Resume Comparaison de quelques proprietes du facteur CAMP de Streptococcus agalactiae avec l'exosubstance active hemolytique latente de Streptococcus uberis Des exosubstances hemolytiques latentes peuvent etre obtenues sous forme prepurifiee par precipitation avec de l'acetone aussi bien a partir de Streptococcus agalactiae (facteur CAMP) qu'a partir de Streptococcus uberis (designe comme facteur UBERIS). Les deux exosubstances provoquent une hemolyse complete des erythrocytes de mouton dans la zone de la toxine beta des staphylocoques. Une telle hemolyse a ete observee seulement avec le facteur CAMP dans la zone de l'exotoxine de Corynebacterium pseudotuberculosis. Ce phenomene decrit pour les facteurs CAMP et UBERIS dans la zone de la toxine beta est inhibe par le ou les facteurs presents dans des serums sanguins foetaux ainsi que dans des serums sanguins hyperimmunises. Les facteurs CAMP et UBERIS sont immunogenes et reagissent specifiquement en double diffusion sur gel avec des antiserums homologues. Les deux exosubstances possedent des proprietes antigeniques differentes et se differencient egalement de la toxine delta des staphylocoques. Ainsi s'offre une possibilite de serotypisation supplementaire entre Streptococcus agalactiae et Streptococcus uteris CAMP-positif. Resumen Comparacion de algunas propiedades del CAMP-factor de Streptococcus agalactiae y de una exosubstancia de Streptococcus uberis con accion hemolitica latente Mediante la precipitacion por acetona se pueden conseguir, en la forma prepurificada, las exosubstancias con la propiedad hemolitica latente, tanto de Streptococcus agalactiae, que se suele nombrar CAMP-factor, como de Streptococcus uberis, que se designo UBERIS-factor por los autores del articulo presentado. En la zona de accion de beta-toxina estafilococica las dos exosubstancias mencionadas producen hemolisis completa de los eritrocitos de oveja, mientras en la zona de accion de la exotoxina de Corynebacterium pseudotuberculosis se observa la hemolisis completa solo en el area del CAMP-factor. Los sueros fetales, normales e hiperinmunes contienen un factor (o factores) capaz de inhibir el fenomeno descrito tanto para CAMP-factor, como para UBERIS-factor, en la zona de beta-toxina estafilococica. CAMP-factor e igual UBERIS-factor son inmunogenos y reaccionan especificamente en la precipitacion en agar con los antisueros homogeneos. Las dos exosubstancias estreptococicas difieren antigenicamente una de la otra y a la vez las dos difieren de la delta-toxina estafilococica, lo que ofrece una posibilidad para suerotipificacion complementaria de Streptococcus agalactiae y de las cepas CAMP-positivas de Streptococcus uberis.

Studies were made on the morphological variety in plaques produced by phage lysis and autoplaques in Pseudomonas aeruginosa strains. The great variability of phage plaque morphology may lead to a confusion with the autoplaques produced spontaneously by P. aeruginosa strains. The production of autoplaques is characteristic of a large number of clinical strains of P. aeruginosa. The appearance of autoplaques may complicate analysis of significant clinical strains of P. aeruginosa.

Nine phages of coagulase-negative staphylococci were studied carefully with regard to their electron microscopic morphology. Two different groups of phages were detected: the larger phages showed heads of 121 nm diameter on average and tails with 229 nm length and 23 nm width. The smaller phages were characterized with heads of about 76 nm diameter and tails of about 177 nm length and 13 nm width (negatively stained with KPT).

Concerning the chemical induction of lysogeny of Staphylococcus aureus only little is known. Therefore, we performed induction experiments on 43 Staph, aureus strains using nine mutagenic substances and 15 chemotherapeutics. Only three of the nine mutagenic substances (beta-propiolacton, nitrosoguanidin and mitomycin C) and six of the 15 chemotherapeutics (penicillin G, ampicillin, cephalothin, nebacetin, novobiocin and nalidixie acid) showed inducing effects. Our results demonstrated furtheron that the lysogeny inducing ability is dependent on strains and substances used.

Immunological skin tests were carried out in 213 weanling piglets at the weight of 5 to 30 kg. Immediate hypersensitivity was evaluated by help of diagnostic allergens (mould, yeast and bacterial ones), delayed hypersensitivity by help of staphylococcus lysate, cellular immunity (tests de novo) by help of phytohemagglutinin and as common recall antigens were used tuberculin, toxoplasmin, candidic and tetanic antigen. The above substances were applied intradermally to the back of the piglets. For an evaluation of non-specific inflammatory response, sodium lauryl sulphate applied epicutaneously was used. The average reaction to diagnostic allergens (50 PNU in 0.05 ml) evaluated after 20 minutes was characterized as light dermal reactions (the papule size of 3-5 mm); in 22% of piglets moderate dermal reactions (the papule larger than 6 mm) to the diagnostic bacterial allergen (Staphylococcus aureus, Staphylococcus epidermidis) were determined. After an intradermal implantation of histamine (50 micrograms in 0.05 ml) the average size of the papule was 13 mm in 20 minutes. After the application of 0.1 ml of staphylococcus lysate (STAVA), the induration exceeding 10 mm was observed in 13% of piglets in 24 hours. After the application of phytohemagglutinin (100 micrograms in 0.1 ml), the induration exceeding 5 mm in 24 hours was determined in 93% of piglets and in 48 hours in 59% of piglets. Common recall antigens were applied at the volume of 0.1 ml. Forty-eight hours after the application no palpable induration was determined: in 91% of piglets after the application of tuberculin (2 TU PPD in 0.1 ml); in 75% of piglets after toxoplasmin (according to PNY 30-33-74); in 98% of piglets after candidic antigen (100 PNU in 0.1 ml) and in 86% of piglets after tetanic antigen implantations (0.03 Lf in 0.1 ml). The epicutaneously applied sodium lauryl sulphate (at 2.5% and 5% concentrations) caused no inflammatory dermal reactions (erythema or induration) after 24 hours. In piglets it is best to apply phytohemagglutinin for evaluating cellular immunity, staphylococcus lysate for evaluating delayed hypersensitivity to staphylococci and histamine for obtaining the information on a disposition of piglets to allergic diseases.

The authors have modified the one-plate method for the detection of staphylococcal hemolysins. They recommend to use in this method a prepurified form of staphylococcal beta-toxin and of streptococcal CAMP-factor instead of the exclusively beta-tonin-producing strain of Staphylococcus aureus and instead of the intensively CAMP-test positive Streptococcus agalactiae strain, respectively. The authors determined concurrently staphylococcal hemolysins, using a three-plate method in which alpha-antitoxin was employed, to ensure a better evidence of alpha-toxin. A total of 494 staphylococcal strains were examined by both methods. Of this number, 446 Staphylococcus aureus strains were of diverse host origin and 48 were coagulase-negative staphylococcal strains. On the basis of the various hemolytically active staphylococcal toxins, the authors recommend the suggested modification of the one-plate method for their routine detection.

Acute pseudomonas infections require treatment with antibiotics producing a bactericidal effect. The most useful are gentamicin, tobramycin, sisomicin and polymyxin B. In resistant strains, amikacin is indicated in addition. Carbenicillin, ticarcillin, carfenicillin or azocillin should never be given alone but in combination with some of the above preparations. Other drugs, such as chloramphenicol, tetracycline or streptomycin, though effective in vitro, should be avoided. Chemotherapy may be complemented by passive immunization either with hyperimmune specific gama globulin or hyperimmune plasma. A programmatic item of combined treatment is active immunization, especially with toxoid vaccine. Chronic processes are not, perhaps with the exception of urinary infections, suitable for antibiotic therapy. For this reason effective polyvalent vaccines should be developed from appropriate strains. It is now certain that in infections caused by mucous strains (most frequently encountered in cystic fibrosis) the vaccine should be prepared from these strains, since they have distinct functional and antigenic characteristics.

The use of Corynebacterium pseudotuberculosis or of its hemolytically active exosubstance for the detection of staphylococcal hemolysins represents a suitable alterative to the use of the beta-toxin producing strain of Staphylococcus aureus, or of isolated beta-toxin. An increased capture of beta-toxin production was recorded when the CAMP-factor of Streptococcus agalactiae was used. A total of 446 Staphylococcus aureus strains and 48 coagulase-negative staphylococal strains were tested against the prepurified form of hemolytically active exosubstance of Corynebacterium pseudotuberculosis and against the CAMP-factor. The results obtained are in agreement with the testing by means of staphylococcal beta-toxin and of the CAMP-factor. The two modifications of the method for the detection of staphylococcal hemolysins using one plate as proposed by the authors give good results. Their ability of assessment, however, is higher by one degree when the exocubstance of Corynebacterium pseudotuberculosis is employed.

Our phage-set, published in 1975 (6) was modified in that other staphylococcal hoststrains were found more suitable for the propagation. This new typing set of 15 phages should replace our old phage-sets. In comparing the phage-sets of Dr. Verhoef, Dr. Parisi and Dr. Blouse with our phages, the advantages of our new phage-set could be demonstrated. Lysogeny induction experiment with mitomycin C and UV-rays showed all staphylococcal host strains to be lysogenic. Conclusions of the studies performed were derived from and discussed.

Ps. aeruginosa strains--a frequement resuet of an irresponsible antibiotic therapy--represent a common agent of nosocomial infektions. At the same time, gravity of Pseudomonas diseases is also increasing. Lysogeny, bacteriocinogeny and frequent occurrence of autoplaques are the lytic manifestations of Pseudomonas aeruginosa strains which play a great role in the complexity of solving diagnostic, epidemiological end therapeutical problems connected with infections induced by these microbes. A survey is presented of the importance and utilization of the lytic properties of bacterial strains of Pseudomonas aeruginosa during the differentiation, epidemiological typing and further expansion of therapeutical possibilities in infections caused by Pseudomonas aeruginosa bacteria.

Versuche uber Inaktivierung von Staphylokokkenphagen durch Hydroxylamin. Das Resultat wird auch durch die Wahl des Wirtsorganismus bestimmt.

Coagulase-activity is today accepted as the best in vitro-test for proof of staphylococcal pathogenicity. Coagulase-positive staphylococci are considered to be pathogens, coagulase-negative staphylococci as non-pathogens.

The purpose of our study was to check the possibilities of differentiating coagulase-negative staphylococci with the help of biochemical and morphological characteristics. Altogether 59 reactions were examined on a material of 147 strains, which were isolated from human pathogenic processes in USA, Canada, Denmark, CSSR and Germany.

SummaryThe inactivating action of hydroxylamine was studied with representatives of several serological groups of Staphylococcus aureus. It has been found that sensitivity of these phages depends to some extent on the serological group of the phage. For serological group A, however, distinct differences in sensitivity to hydroxylamine have been found. It has been verified that these differences within a given serological group depend on the phage group, i.e. on host bacteria; investigation of polyvalent phages was not made. No direct dependence of phage sensitivity on the base content (G—C) and sensitivity to hydroxylamine was found. For these phages no anomalous base was found. All of the above phages contain the bases G—C and A—T in their DNA molecules.

Lysogeny after Mitomycin C- and UV-induction was studied on 127 pathogenic strains of the speciesStaphylococcus albus. A lysogeny indicator set of 24Staphylococcus albus strains was used. All 127 examinedStaphylococcus albus strains could be shown to be lysogenic, sometimes even multilysogenic.

SummaryWe studied the influence of the growth of polyvalent Staphylococcus phage A/5 in eight bacterial hosts on the course of inactivation of the resultant phage lysates by u.v.- and gamma-radiation and by acridine orange. The starting titrations of all the phage stocks used were identical. The propagation of phage A/5 through various hosts affected the course of inactivation of the phage and, especially, its resistance to u.v.-radiation was increased. Propagation of phage A/5 through host strain B1 produces its resistance both to u.v.- and to gamma-radiation as well as to photodynamic inactivation, whereas the host strains producing the greatest sensitivity of phage A/5 to these three agents, differ in all cases.