[The phenotypic properties of freshly isolated strains of Bordetella]. / Izuchenie fenotipicheskikh svoistv svezheizolirovannykh shtammov bordetell.

As the result of our investigations, newly isolated B. pertussis and B. bronchiseptica strains were studied. The results of these investigations showed that B. pertussis strains isolated under the conditions of immunoprophylaxis were characterized by sufficient stability of the main phenotypical properties which determined their pathogenicity: B. pertussis toxin, fimbrial agglutinogens and filamentous hemagglutinin. At the same time B. bronchiseptica strains isolated from animals proved to be phenotypically variable both in vivo and in the process of in vitro passage.

Bacteriophages of Bordetella sp.: features of lysogeny and conversion.

It has been the purpose of this paper to study molecular-biological features of the Bordetella bacteriophage interaction with the host cell during lysogeny and conversion as well as to determine the degree of homology between genomes of homologous and heterologous bacteriophages. Genomes of bacteriophages from B. pertussis 134, 41405 and B. bronchiseptica 214 were studied. Heteroduplex and restriction analyses revealed a heterogeneity of bacteriophage populations, and their DNAs were found to differ in size and position of inserts. As shown by blot hybridization, the bacteriophage genome is not inserted into the chromosome of the lysogenic cell but apparently exists as an autonomous plasmid replicon. It has been established that during conversion only a part of the phage genome is inserted into the chromosome of the recipient cell.

[Isolation and characteristics of the bacteriophage from the vaccine strain Tohama phase I]. / Vydelenie i kharakteristika bakteriofaga iz vaktsinnogo shtamma Tohama I fazy.

Some properties of bacteriophage phi T isolated from the vaccine strain Bordetella pertussis Tohama phase I and propagated in Bordetella parapertussis 504 cells are presented. Phage phi T belongs to the IV group in accordance with Tikhonenko classification. The diameter of head and length of noncontractile tail sheath are 49.5 +/- 0.5 and 145 +/- 7 nm, respectively. Diameter of the tail sheath is 3.2 +/- 0.6 nm. Molecular mass of the phage DNA is 37 +/- 3 kb. Population of phi T phage is polymorphous and consists of particles the genomes or which vary from each other by the "insert" located 6.8 +/- 0.6 kb from the end of molecule. The blot hybridization has demonstrated that the bacteriophage genome is not inserted into the chromosome of the lysogenic strain. Autonomous location of the phage genome in the host cell is suggested. The temperature and hydrogen ions concentration effects on bacteriophage phi T stability were studied. The conditions for phage suspension storage are described.

[Limitation of phage multiplication by microbes of the genus Bordetella]. / Ogranichenie razmnozheniia fagov mikrobami roda Bordetella.

Possible causes limiting the multiplication of Bordetella phages or inducing their restriction, such as the influence of lysogenic immunity and the restriction-modification (R-M) system or the incompatibility of the receptor apparatus, have been studied. The limitation of the multiplication of phages by some B. bronchiseptica and B. pertussis strains has been shown to be due to the presence of the R-M system and lysogenic immunity. In five B. bronchiseptica strains and two B. pertussis strains site-specific endonucleases (restrictases) with Hind III specificity have been detected. One B. bronchiseptica strain without the R-M system has been detected. B. bronchiseptica strains producing site-specific endonucleases are practically nonpathogenic for humans, grow in common culture media and selectively produce only one restrictase, type Hind III, which guarantees from the admixture of other specific endonucleases. The B. parapertussis strains under study (altogether 100 strains) have not been found to limit the multiplication of Bordetella test phages. The absence of site-specific endonucleases has also been confirmed biochemically. These strains are recommended as indicator strains for the multiplication of Bordetella phages.

Changes within the genus Bordetella influenced by phages.

Bordetella phages give rise to changes in the important phenotypic properties of all three Bordetella species. The changes depend on the phage system used. The treatment of Bordetella parapertussis with phages from B. pertussis led to toxic clones with manifold combinations of agglutinogens specific for all three Bordetella species. Under the influence of B. pertussis phages clones are formed which differ phenotypically from the recipient strain. Some of these clones are sensitive to some phages. In addition, the converted strains were able to propagate in the brain tissues of mice and gave rise to paralytic symptoms or death. Adhesive and hemagglutinating activities were detected in the converted strains and biochemical properties of the converted strains changed in comparison to the recipient strain. The sensitivity of converted strains to antibiotics was investigated and the tolerance of converted strains to erythromycin was used as a marker of conversion. As to their most striking features the converted strains resembled wild strains of B. bronchiseptica isolated from rabbits. While it was not possible to show the ability of pertussis phages to propagate in strains of homologous species by the agar layer method, pertussis phages did adsorb maximally on strains of B. pertussis.

[Detection of Bordetella bronchiseptica strains containing the main agglutinogens of all species of the genus Bordetella]. / Obnaruzhenie shtammov Bordetella bronchiseptica, soderzhashchikh glavnye aggliutinogeny vsekh vidov roda Bordetella.

The study of 26 B. bronchiseptica strains with typical morphological and biochemical properties resulted in the detection of 8 strains having the main specific agglutinogens of 3 Bordetella species (serovars) in different combinations. The presence of the agglutinogens was confirmed in the agglutination test and the agglutinin adsorption test with the use of monospecific antisera to the main agglutinogens. The comparison of natural B. bronchiseptica serovars and artificial convertants (resulting from the conversion of B. parapertussis by B. pertussis phages) revealed their identical biochemical activity, their capacity for causing necrosis when injected intradermally into rabbits and for the formation of two types of colonies, differing in size and serological activity. In contrast to B. parapertussis convertants, B. bronchiseptica serovars had no lysogenic properties and were sensitive to B. pertussis and B. bronchiseptica phages.

[Modern bacterial vaccines]. / Moderne bakterielle Impfstoffe.

A well founded vaccination strategy is of crucial importance for controlling communicable diseases. The WHO Enlarged Vaccination Programme of 1976 provides to protect by vaccination all children in the world against six infection diseases - diphtheria, pertussis, tetanus, measles, poliomyelitis, and tuberculosis, a sure immunization prophylaxis being possible against them. The perspectives of further development of some classical vaccines as against cholera, typhoid fever, and pertussis are reviewed. Some recent bacterial vaccines are discussed being of special significance for controlling nosocomial infections and for protecting patients in intensive therapy and/or persons with lowered immunological defence. New ways for production of vaccine matters are shown such as capsule polysaccharides, membrane proteins, subcellular fractions, and vaccines with synthetic carrier molecules. Fundamental problems concerning the initiation of vaccination and the demands for an enlarged proof of innocence of vaccines are discussed.

[Conversion of Bordetella parapertussis serovar through lysogeny produced by pertussis phages]. / Konversion des Serovars von Bordetella parapertussis durch Lysogenie, erzeugt mit Pertussisphagen.

Bacteriophages from Bordetella pertussis were titrated on the indicator strain B. parapertussis 17903 by using standard soft agar techniques. Secondary growth, occasionally observed in some phage plaques, was isolated and transferred onto selective media. Judging from growth on these special media and microscopic examination the isolated clones consisted entirely of Bordetellae. Determination of the agglutinogen pattern of 160 of these clones revealed that 88% contained agglutinogen 1; 87.5% agglutinogen 14, and 80.1% agglutinogen 12 (Table 3). However, none of the strains expressed the agglutinogen pattern of either the phage donor or the recipient strain. The isolated clones were lysogenic as demonstrated by phage production and immunity against superinfection (95% of the clones).--Absorption of the monospecific antisera with whole cells from lysogenic strains resulted in a drastic decrease or even complete loss of specific antibodies towards those antigens identified by slide agglutination reactions (Table 4). Cross absorption experiments with antisera against B. pertussis, B. parapertussis and strain 73 1/7 and various strains of the genus Bordetella and with a number of lysogenic strains showing various agglutination patterns allowed the conclusion that the latter ones were serologically related to B. pertussis. The lysogenic strains completely absorbed antibodies against B. bronchiseptica, and those strains that carried the agglutinogen 14 also absorbed antibodies against, B. parapertussis (Table 5). In conclusion, these results support the necessity to revise the subdivision of the genus Bordetella into three species. A change of B. parapertussis to B. pertussis within the epidemiological processes is considered.

[Toxigenicity conversion by pertussis phages in Bordetella parapertussis]. / Konversiia toksigennosti kokliushnymi fagami u bordetella parapertussis.

For the first time toxigenicity conversion in B. parapertussis induced by B. pertussis phages was discovered. The clones of B. parapertussis recipient strain No. 17903 used in this study were subjected to lysogenization with 4 B. pertussis phages; as a result, 95% of these clones became immune to the repeated phage infection, developed spontaneous phage production and showed toxic properties (lethal toxicity due to the action of thermolabile and thermostable toxins) characteristic of the donor strains from which B. pertussis phages had been obtained. Differences in the degree of toxicity shown by the converted strains were determined by means of the spleen index. The convertants thus obtained did not possess protective potency.

[Characterisation of major biological properties of Bordetella bacteriophages]. / Kharakterstika osnovnykh biologicheskikh svoistv bakteriofagov Bordetella.

The main biological properties (morphology of negative colonies, parameters of adsorption and single development cycle) of B. pertussis and B. bronchiseptica phages, isolated spontaneously and by induction with mitomycin C, were studied. To compare these characteristics, one B. parapertussis indicator strain was used, and the experiments were carried out under identical conditions. Highly active sera were obtained with the use of complete Freund's adjuvant. B. pertussis phages isolated from the strains of different serovars were serologically related, but not identical, and differed in their constant characterizing their rate of neutralization with homologous antisera. The adsorption of the phages on homologous strains was more intensive than on the cells of B. parapertussis indicator strain. However, the authors failed to observe the further development of the phages in the host cells.

[Transparent solid nutrient medium for studying the lytic spectrum of bacteriophages of microbes of the genus Bordetella]. / Prozrachnaia plotnaia pitatel'naia sreda dlia izucheniia liticheskogo spektra bakteriofagov mikrobov roda Bordetella.

A solid, transparent culture medium for the study of the lytic spectrum of the phages, active against B. pertussis and B. bronchiseptica, in respect to homologous and heterologous bacteria of the genus Bordetella has been developed. The Cohen-Wheeler liquid medium with nicotinic acid and nicotinamide added, solidified with agar, is nicotinamide added, solidified with agar, is used as the base of the new medium. This base ensures the growth of B. parapertussis and B. bronchiseptica. To stimulate the growth of B. pertussis, the tissue stimulant of B. pertussis growth (a transparent substrate obtained from the tissue of the large intestine of a rabbit) has been used. With 10% of this stimulant added, B. pertussis cells have been found to preserve their typical morphological and immunobiological properties.

[Effect of certain factors on the variability of strains of pertussis microbes]. / Vliianie nekotorykh faktorov na izmenchivost' shtammov kokliushnogo mikroba.

The population of degraded cells having stable changes in some phenotypical properties were isolated after subculturing some laboratory Bordetella pertussis strains in Bordet-Gengou culture medium and casein-charcoal agar with blood, treated with mitomycin C and allowed to proliferate in the spleen of mice injected intravenously with microbial suspensions. The characteristics indicative of cell degradation were the growth of large yellowish-white colonies appearing in 24 hours, the destruction of the agglutinogenic complex and toxic substances causing the atrophy of the spleen in mice, the increased capacity for active proliferation in the spleen. Electron-microscopic study revealed that the variants obtained by subculturing in culture media had the damaged membrane with the formation of cell-wall invaginations having rounded membrane-like formations on their surface, disappearing after treatment with mitomycin C; the treatment of the initial strains with mitomycin C resulted in cytoplasmic damage with the coagulation of the nucleoid.

[Bordetella pertussis bacteriophage]. / Bakteriofag Bordetella pertussis.

For the first time Bordetella pertussis bacteriophage was isolated, and its presence was confirmed by electron microscopy and by agar layer titration. The lysogenic strains were activated by their treatment with mitomycin C in a dose of 4.5 mg/ml. The phage system of the Bordetella genus, heretofore unknown, has been revealed: Bordetella pertussis phage lyzed all the tested strains of Bordetella parapertussis (25 strains) and could be passaged in these strains. The phage formed turbid and transparent negative colonies 0.1 mm and 0.15 mm in size. The phage titer (e. g., in strain No. 3865) was 1 X 10(10). The lysogenic variants of Bordetella pertussis, capable of spontaneous release of the phage, were obtained. These variants were characterized by changes in some of their phenotypical properties, e.g., the increased content of certain toxic substances and increased virulence.