Specific Identification of Fraction I-Positive Pasteurella pestis Colonies on Antiserum-Agar Plates

A method is described for the use of antiplague serum in Blood Agar Base plating media to detect fraction I-positive Pasteurella pestis. The antiserum was produced conveniently and in large volume in rabbits by use of Cutter plague vaccine combined with Freund's complete adjuvant. P. pestis colonies were specifically identified within 48 hr after plating by the presence of a precipitin ring surrounding each colony. The basis of the test was shown to be a precipitin reaction between fraction I antigen released from P. pestis colonies after chloroform vapor treatment and fraction I antibody present in the antiserum-agar medium.     

Immunogenicity of B-antigen in experimental plague and pseudotuberculosis

The results of the evaluation of the immunogenic properties of B-antigen, earlier identified in the culture fluid of Yersinia pseudotuberculosis submerged culture, with respect to experimental plague and pseudotuberculosis are presented. B-antigen has been shown to produce protective effect in guinea pigs and, probably, hamadryas baboons, but not in white mice infected with the causative agent of plague. Immunizaton with B-antigen protects guinea pigs from primary pneumonic plague caused by both capsule-forming and noncapsular Y. pestis virulent strains. Passive immunization with antibodies to B-antigen induces limitedly pronounced protective effect in guinea pigs and is not effective for white mice with respect to experimental plague. No active or passive protection of white mice or guinea pigs, infected with Y. pseudotuberculosis cultures, has been achieved by the injection of B-antigen or antibodies to it.     

Protein expression of Yersinia pestis during changes due to long-term cultivation in vivo]

The electrophoretic study of Yersinia pestis proteins made possible to find the significant modification of Yersinia pestis polypeptide specters when the bacteria were cultivated in semi-penetrable cells implanted into the guinea pigs peritoneum. The proteinogramms of the isolates from the implanted cells lacked the stained bands characteristic of Yersinia pestis cells grown in vitro and contained the new polypeptides absent from the bacteria grown on the Hottinger agar plates. The difference was found at the late stage of bacteria incubation in implanted cells and had the predominantly reversible characteristics. The protein of Yersinia pestis being changed in vivo is proposed to be the species specific fraction I.     

Pesticinogeny: a Characteristic Useful for Presumptive Identification and Isolation of Pasteurella pestis

Current methods of identifying Pasteurella pestis rely heavily on tests specific for detecting fraction I, the envelope antigen. Pesticin I, a bacteriocin inhibitory for P. pseudotuberculosis, has been demonstrated in nearly all tested strains isolated from human infections. The results of using this characteristic as an identifying trait for P. pestis were compared with results reported for detecting fraction I by fluorescent-antibody and antiserum-agar techniques. Data indicate that, although certain atypical strains of P. pestis fail to react in one system or the other, a combination of these tests provides positive identification in all cases. Detection of P. pestis in contaminated materials is greatly facilitated, and the simplicity of this test makes it a valuable tool in the study of plague infections and an important adjunct to methods currently in use. The use of the pesticin I assay is not intended to replace other accepted techniques, but rather to supplement them and increase the effectiveness of plague investigation.     

Changes in the "latent" virulence of a vaccinal strain of Yersinia pestis multiplying within macrophages

The multiplication of Y. pestis vaccinal strain inside peritoneal macrophages of guinea pigs and white mice in vitro leads to an essential increase in its latent virulence. This effect is most pronounced when guinea pig macrophages are used. Changes in the latent virulence of Y. pestis vaccinal strains, occurring in the process of their passage inside macrophages in vitro, correlate with those observed in vivo, i.e. in animal experiments.     

The effect of a plague pathogen plasmid on lethality and immunogenicity

On the model of Yersinia transconjugants (Yersinia pestis, Yersinia pseudotuberculosis, Yersinia enterocolitica) carrying the conjugative cointegrates containing the 47 and 65 Md plasmids from Yersinia pestis the data were obtained on the different affects of the latter plasmids on the lethality and immunogenicity conferred by the host bacterial cells. The plasmid effects were drastically different during bacterial infection in mice or guinea pigs. The possibility of appearance of the recombinant Yersinia in natural epizootic foci of plague and suggestions on their regulating role are discussed.     

Role of histamine receptors of mouse and guinea pig peritoneal leukocytes in the pathogenetic effect of Yersinia pestis adenylate cyclase]

The effect of purified preparation of adenylate cyclase isolated from Y. pestis on peritoneal leukocytes of white mice and guinea pigs was investigated. Y. pestis adenylate cyclase was shown to accomplish its pathogenic action via histamine-specific receptors on the surface of eukaryotic cells. The involvement of H1 and H2 histamine receptors on target cells in the adenylate cyclase action leading to development of plague infection is discussed.     

The significance of pesticin 1 for the virulence and immunogenicity of Yersinia pestis]

The work deals with the study of the virulent and immunogenic properties of Y. pestis strains which lost their capacity for producing pesticine 1 as the result of the insertion of a Tn-like element into the 6-MD plasmid responsible for this property. The "switching-off" of gene pst induced a decrease in the virulence of Y. pestis injected subcutaneously into white mice and guinea pigs and had no influence on its level of immunogenicity for white mice. A suggestion was made that pesticine 1 played no essential role in the expression of the virulence and immunogenicity of Y. pestis penetrating into the body by subcutaneous route.     

The phagocytic activity of peritoneal macrophages in relation to Yersinia pestis with defective and complete Fra genes

In vitro study of phagocytosis has shown that in guinea pigs fraction 1 is conducive to the ingestion of Y. pestis by macrophages, to survival and proliferation of Y. pestis cells in these macrophages, as well as to their specific transformation leading to their increased ingestive and bactericidal activity with respect to Y. pestis. In mice the role of fraction 1 in phagocytosis has proved to be less significant.     

Reaction of macrophage disappearance as a method for the evaluation of cellular immunity in experimental plague.

The reaction of macrophage disappearance (RMD) was found to develop one day after the immunization of experimental animals with the antigen FIA of Y. pestis and to persist for 21 days, correlating with a pronounced protective effect of the above antigen over this period. There were differences between white mice and guinea pigs in the intensity of the RMD, particularly when immunization was carried out using an FIA preparation without prolonged action. These differences are thought to reflect varying degrees of immunity elicited by FIA immunization. It is recommended that the RMD be used to quantify cellular immunity to Y. pestis.     

The effect of the adhesion pili of Yersinia pestis on the physiological activity of the leukocytes and macrophages in experimental animals

The data on the influence of the preparation of Y. pestis adhesion pili on peritoneal macrophages in white mice and guinea pigs are presented. Y. pestis adhesion pili have been found to induce the dose-dependent increase of cell chemiluminescence. They have also been found to induce a number of biochemical changes in target cells: the secretion of myeloperoxidase, an increase in the activity of cAMP-dependent protein kinases.     

Immunogenicity of Yersinia pestis grown on nutrient media at 28 and 37 degrees C

The immunogenicity of Y. pestis strain EV, grown in yeast-casein medium, yeast medium with Hottinger digest and yeast medium with sunflower-seed protein at 28 degrees C and 37 degrees C, for guinea pigs and white mice has been studied. As revealed in this study, these media ensure the formation of highly immunogenic populations of Y. pestis strain EV and, therefore, can be used for growing Y. pestis vaccine strains. Considerable fluctuations in the content of such highly protective antigen as fraction 1 do not affect the immunogenicity of live cultures of Y. pestis strain EV. This is due to the leveling of differences in the content of this antigen in the process of the multiplication of these bacteria in laboratory animals.     

Duration and intensity of postvaccinal immunity to plague in experimental animals]

Experiments were conducted on guinea pigs and Papio hamadryas; it was shown that a reduction of the intensity of postvaccinal immunity occurred at various periods after a single vaccination. In inhalation method of immunization in guinea pigs it decreased in 6 months 135 times, in monkeys in one year--133 times. However, at the mentioned periods vaccination provided protection of 50% of the animals from infection with Past. pestis in a dose constitutin 20 to 25 aerosol LD50 for nonimmunized animals. Despite the more pronounced (57--640 times) reduction of the intensity of immunity than in the animals vaccinated by inhalation, in the subcutaneously vaccinated guinea pigs in subcutaneously infected with Past. pestis protection level remained high (resistance index in 3 and 6 months constituted 37.10(6) and 3-3-10(6), respectively).     

鼠疫动物模型:经典与问题

一直以来,鼠疫菌的研究多围绕致病机制、疫苗效果评价及鼠疫治疗方案选择展开,而合理有效的鼠疫动物模型正是这些实验研究的基础与前提。研究者构建了包括小鼠、豚鼠及非人灵长类模型在内的多种动物模型,而这些模型也为人类认识、防御及治疗鼠疫菌提供了帮助,特别是一些经典动物模型沿用至今。但随着研究的深入,有些动物模型由于存在某种问题而逐渐被淘汰。本文就鼠疫菌动物模型构建以来,不同模型的优势与不足及其应用展开综述,以期为研究者提供参考。     

Evaluation of the modulating action of Yersinia pestis adenylate cyclase on guinea pig peritoneal leukocytes using chemiluminescence

The biological action of Y. pestis adenylate cyclase on peritoneal leukocytes of guinea pigs has been studied by means of chemiluminescence. Y. pestis adenylate cyclase is supposed to contribute to the "oxidation" explosion of phagocytes in plague.     

The phenomenon of pilus formation in the interaction of Yersinia pestis with macrophages in experimental animals

According to the data of the enzyme immunoassay, adhesion pili were expressed by Y. pestis cells EB76 after their cultivation in media with pH similar to that of phagolysosomes. The expression of adhesion pili was found to occur 18 hours after the contact of apiliate cells EB76 with a monolayer of native, but not inactivated (at 56 degrees C for 30 minutes) macrophages of the peritoneal exudate of white mice and guinea pigs. Purified adhesion pili possessed cytotoxic action and inhibited the digestive activity of macrophages with respect to Y. pestis. The formation of pili in interaction with macrophages and the pronounced effect of the preparations of purified pili on the function of phagocytes make it possible to regard the formation of pili as an important determinant of virulence.     

The isolation and characteristics of variant plague microbes not producing the capsular antigen

Fifteen stable variants of Yersinia pestis strains exhibiting different degrees of virulence for white mice and guinea pigs were obtained. Multiple passages of the organisms at 37 degrees C in a fluid nutrient medium containing antiplague agglutinating serum were found to be the most efficient method for obtaining noncapsular forms of the plague agent. Acquisition of the Fra(-) phenotype both by wild and laboratory strains was not associated with a loss of the high-molecular plasmid by cells but was probably a result of mutational alterations in the plasmid genes.     

Virulence of Venezuelan Equine Encephalomyelitis Virus Subtypes for Various Laboratory Hosts

Mice, guinea pigs, and duck embryo cell cultures were inoculated with known subtypes of Venezuelan equine encephalomyelitis (VEE) virus and the attenuated (TC-83) strain of VEE. With the exception of TC-83, all strains were highly pathogenic for suckling mice by either intracranial or intraperitoneal routes of inoculation used. Virulence for older mice and guinea pigs provided a means to distinguish strains. In addition, virulence or lack of virulence for adult mice or guinea pigs provides a rapid method for separating epizootic subtype IB from TC-83 VEE virus isolates.     

Evaluation of hypoxic state in experimental animals induced by Yersinia pestis antigens

Some characteristics of purine metabolism in experimental animals (white mice, clawed jirds and guinea pigs), injected intraperitoneally with Y. pestis "murine" toxin and capsular antigen (Fraction 1), were studied. Under the influence of sublethal doses of these antigens increased levels of guanine and xanthine in blood were noted. Changes in the content of xanthine oxidase in cells were insignificant. In white mice and clawed jirds the activity of succinate dehydrogenase decreased under the action of "murine" toxin and increased after the injection of Fraction 1.     

Yersinia pestis L-forms in rodents and ectoparasites

Y. pestis L-forms and bacterial forms persist in the body of great gerbils for 40 days. L-forms are poorly phagocytized and can persist in phagocytes for a long time. In guinea pigs immunized with vaccine EV, Y. pestis antigen could be detected till day 160. An unstable L-form was isolated from Ornithodoros mites 3 years after their experimental infection with Y. pestis. Bacterial forms persist in mites for 1-3 years. For 5 years Y. pestis antigen is regularly detected in a high percentage of mites.