Effect of subinhibitory concentrations of antibiotics on catalase activity of plague microbes]

It was shown that the presence of subinhibitory concentrations of ampicillin, cefotaxime or gentamicin in the cultivation medium had a marked inhibitory effect on the catalase activity of plague microbe. The effect depended on the characteristic features of plague microbe strains and the incubation temperature. When the cells of a virulent strain of the plague microbe Y. pestis 1300 were cultivated at a temperature of 37 degrees C on a medium containing the subinhibitory concentrations of ampicillin or cefotaxime, the pathogen virulence for albino mice significantly decreased.     

Chromosomal resistance of plague agent to quinolones]

The nature of increasing chromosomal resistance to quinolones was studied in a model of the plague microbe. Five virulent strains of the natural plague microbe (Y. pestis) were used in the experiment: 363 (1/1479), 231, 2385, 2442 and 2444. The one-stage procedure for isolation of the mutants was applied. It was shown that the frequency of the one-stage mutants resistant to oxalinic acid, pefloxacin and ciprofloxacin amounted to 10(-9)-10(-11) and was 2 to 3 orders of magnitude lower than that of the mutants resistant to nalidixic acid. Two types of the plague microbe mutants resistant to the quinolones were detected: those resistant to the quinolones to the generations (Nalr-phenotype) and those resistant to the representatives of the 3rd generation quinolones (Nals-phenotype). The quinolones were not efficient in the treatment of albino mice with experimental plague caused by the quinolone-resistant forms of the plague microbe.     

Study of trimethoprim sensitivity in strains of Yersinia pestis isolated from various natural foci of plague

The effect of exogenic addition of purines and pyrimidines on trimethoprim sensitivity was studied in 134 vaccinal and virulent strains of the plague causative agent isolated at various periods in 12 natural plague foci. 74 strains proved to be sensitive to low concentrations of trimethoprim in the cultivation media irrespective of the presence or absence of thymine or thymidine in them. In this respect the strains differed from many other bacterial species which in the presence of thymidine or thymine were resistant to high concentrations of trimethoprim. 60 natural arginine deficient strains of the plague causative agent from Transcaucasia and Mongolia showed high levels of resistance to trimethoprim on media with thymine or thymidine. The possible mechanism of the plague microbe sensitivity to trimethoprim in the presence of thymine or thymidine is discussed with an account of the literature and original data.     

Sensitivity of different strains of the causative agent of plague to disinfectants]

Various strains of the plague microbe was found to have statistically significant differences in their sensitivity to phenol, formaldehyde and chloramine. These differences did not correlate with the origin, virulence and nutritional requirements of the plague strains. The sensitivity of the plague microbe to formaldehyde and phenol was found to be related to the permeability of its cell wall.     

Spheroplasts of plague microbe strains from the Transcaucasian uplands and their capacity for pesticin synthesis]

Spherical formations of the plague microbe strains from the Transcaucasian Upland, I plague microbe strain of the sandwort variety and I strain of the marmot variety were obtained under the effect of lithium chloride. They had the remains of the cell wall, were viable, sensitive to osmotic shock, preserved sensitivity to the specific bacteriophage and pesticins. All this was evident of isolation of the spheroplasts of the plague microbe. The spheroplasts showed a capacity for pesticin production. The pesticin synthesis by the spheroplasts of the plague causative agent from the Transcaucasian Upland increased with an increase in the content of lithium chloride in the medium. The largest inhibiiton zones were observed, when 0.7-0.8 per cent of lithium chloride were present in the medium. In the spheroplasts of the plague causative agent from the Mountain Altai (the marmot variety) the pesticin synthesis was inhibited with an increase in the content of lithium chloride in the medium. The activity spectrum of the pesticins of the spheroplasts of the plague causative agent from the Transcaucasian Upland and the spheroplasts of the strains of the marmot and sandwort varieties was broader than that of the rod-like forms of these strains. The indicator properties were found in the strains of the plague microbe of the marmot and sandwort varieties with respect to the pesticins of the spheroplasts of the sel-like producing organisms and organisms from the Transcaucasian Upland.     

Paradoxopsyllus scorodumovi (Aphaniptera) flea, an effective vector of plague in a Gorno-Altai natural focus

A study was conducted on the fleas of P. scorodumovi and five local strains of the plague microbe, one of which is typical of the strains of the Altai subspecies and four are non-typical of this nidus. The fleas of this species are capable to transmit not only the plague agent of the strains typical of this nidus but also non-typical ones which differ in some biological properties and are avirulent for most carriers but Pallas's pika. Biological peculiarities of fleas of P. scorodumovi in addition to their high efficiency as vectors of the plague microbe enable us to associate the more active autumn epizooty with fleas of this species.     

接种不同毒力的马立克氏病病毒后鸡病毒血症的动态比较

1日龄非免疫鸡分别接 马立克氏病病毒（MDV）Ⅰ强毒GA株、Ⅰ型MDV疫苗毒CVI988株和Ⅲ型火鸡疱疹病毒（HVT）疫苗株后第4日起,定期采血并和抗MDV囊膜糖蛋白B(gB)单克隆抗体介导的间接免疫荧光试验检测MDV在外周因液单核细胞（PBMCs）中的感染状况。结果发现,自接种Ⅰ型强毒GA株后第4日至鸡发病死亡前,都能检出GA株引起的病毒血症,并于2周左右达到高峰;自接种CVI988株后第4日至第20日止,能检出病毒血症,并于第8天左右达到高峰;自接种HVT后第4日至第16日止,能检出病毒血症,并于第6天左右达到高峰。与此同时,将GA株病毒血症的IFA检测结果与细胞培养上病毒空班计数试验结果比较,发现IFA试验比空斑计数试验更为敏感。本试验既可用于判断对鸡作MDV疫苗免疫的接种效果,又可用于检测MDV野毒感染状态。     

Role of the Yersinia pestis yersiniabactin iron acquisition system in the incidence of flea-borne plague

Plague is a flea-borne zoonosis caused by the bacterium Yersinia pestis. Y. pestis mutants lacking the yersiniabactin (Ybt) siderophore-based iron transport system are avirulent when inoculated intradermally but fully virulent when inoculated intravenously in mice. Presumably, Ybt is required to provide sufficient iron at the peripheral injection site, suggesting that Ybt would be an essential virulence factor for flea-borne plague. Here, using a flea-to-mouse transmission model, we show that a Y. pestis strain lacking the Ybt system causes fatal plague at low incidence when transmitted by fleas. Bacteriology and histology analyses revealed that a Ybt-negative strain caused only primary septicemic plague and atypical bubonic plague instead of the typical bubonic form of disease. The results provide new evidence that primary septicemic plague is a distinct clinical entity and suggest that unusual forms of plague may be caused by atypical Y. pestis strains.     

Thymidine phosphorylase activity and its relationship to trimethoprim in initial strains and thymidine-, thymine-dependent and trimethoprim-resistant mutants of plague microbe]

The comparative study revealed thymidine phosphorylase activity in the initial strains of a plague microbe of the field variety and in thymidine-, thymine-dependent and trimethoprim-resistant mutants of the plague microbe of other varieties. The data fully conformed to the results of the microbiological investigation of the strains' ability to grow on the nutrient media with trimethoprim in the presence of thymine and thymidine. On the basis of these results it appeared possible to divide the initial and mutant strains of the plague microbe into four arbitrary groups: initial strains of the plague microbe of all the varieties except the field ones sensitive to trimethoprim under any temperature conditions of incubation on any medium with any supplements; initial strains of the plague microbe of the field variety resistant to trimethoprim at 28 degrees C in the presence of thymine or thymidine alone; Tmpr mutants whose resistance to trimethoprim at 28 degrees C did not depend on the presence of thymine or thymidine, purine and vitamins, but depended on the presence of these substances at a temperature of 37 degrees C.     

The effect of different Yersinia pestis antigens on the cellular link in immunity

Immunization with live plague vaccine has been shown to give no protection to thymectomized mice from subcutaneous challenge with Y. pestis virulent strain. Under the action of the vaccine or individual Y. pestis antigens (fraction I) the functional and morphological activation of thymocytes and macrophages is observed, more pronounced in C57BL/6 mice and less pronounced in CBA mice. Y. pestis antigenic preparations (fractions I and II, pesticin) act as T-cell mitogens and are thus capable of inducing the in vitro proliferation of thymocytes. At the same time the in vivo action of fraction II induces a decrease in the level of lymphocytes in the peripheral blood of mice and the destruction of lymphocytes in their thymus and spleen.     

Transport of basic amino acids into Yersinia pestis cells

The transport of the basic amino-acids into the plague microbe cells has been shown to proceed by means of two transfer systems, the common transfer system and the system specific for lysine. Besides that, arginine can be transported into the plague microbe cells by the histidine transport system. The vaccine strain EV NIIEG was used as a model to study the amino-acid transport systems.     

Experimental evaluation of prospects for the use of beta-lactams in plague infection caused by pathogens with plasmid resistance to penicillins]

High therapeutic efficacies of ceftriaxone, ceftazidime, cefotaxime and azthreonam in the treatment of experimental plague induced by beta-lactamase-producing strains of the plague microbe containing R plasmids RP-1, R57b and R40a were shown to correlate with their in vitro antibacterial activities. The therapeutic efficacy of sulbactam/ampicillin was recorded in the treatment of plague induced by the strain containing R plasmids R57b and R40a (the treatment course of 7 days). However, it was lower when the infection was due to the strain containing plasmid RP-1 (beta-lactamase TEM-2). Cefoperazone was not active in the treatment of experimental plague induced by the strains containing plasmids RP-1 and R57b (beta-lactamases TEM-2 and OXA-3). Ceftriaxone versus the antibiotics tested was considered to be the drug of choice for the etiotropic therapy of plague induced not only by the type strains of the plague microbe but also by its variants with the plasmid pattern resistance to penicillins.     

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.     

Antibiotics of the aminoglycoside group (gentamicin, sisomicin and amikacin) in the prevention and treatment of experimental plague]

Activity of aminoglycosides such as gentamicin, sisomicin and amikacin against plague microbe strains of natural origin was studied in vitro. It was also studied in prophylaxis and treatment of experimental plague infection in albino mice. The MAC of gentamicin and sisomicin for 50 strains of the plague microbe was 0.2-1.6 micrograms/ml. For the majority of the strains it was 0.4 micrograms/ml. The amikacin MICs were 0.4-3.2 and 0.8 micrograms/ml, respectively. High efficacy of gentamicin, sisomicin and amikacin was shown in prophylaxis and treatment of experimental plague infection in albino mice. The optimal doses of the antibiotics were determined. Under definite conditions such as the use of short-term regimens and higher intervals, advantages of sisomicin over gentamicin and amikacin in prophylaxis of experimental plague infection were observed.     

The effect of Shigella sonnei strains opposite in their virulence on different links in the immune system]

For the first time different action of S. sonnei strains, opposite in their virulence, on hematopoiesis and the functional activity of T- and B-lymphocytes has been shown. The hematopoiesis-disturbing action of virulent shigellae is manifested by their capacity, more pronounced than similar capacity of an avirulent (vaccine) strain, for stimulating the processes of endo- and exocolony formation, the proliferation of hematopoietic stem cells and their migration to the blood. The effect produced by shigellae on T-cell-mediated immune response is manifested by the suppression of macrophage migration and its subsequent activation, whose manifestations and duration depend on the virulence of S. sonnei strains under study. The modulating effect of S. sonnei on B-cell-mediated immune reactions is manifested by the inhibiting action of S. sonnei virulent strain and the stimulating action of S. sonnei vaccine strain on the formation of antibody-producing cells synthesizing S. sonnei lipopolysaccharide antibodies shortly after the injection of shigellae. The results of this study indicate that S. sonnei virulent and avirulent (vaccine) produce multifunctional and differing effects on cell-mediated immune reactions, these processes being dependent on the virulence of shigellae and their individual specific antigens.     

Characteristics of the multiplication of a virulent strain of the plague microbe in Xenopsylla cheopis fleas infected parenterally

Xenopsylla cheopis fleas infected parenterally with the virulent strain of plague microbe of gerbil variant preserved the agent to the end of their lives. In the body cavity the microbes retained their ability for reproduction which was, however, limited. During the first seven days after the infection the number of microbes slightly increased and later became stabilized. Its mean indices (mean g) varied within the limits of 500 to 2000 microbe cells per 1 individual, maximum index rarely exceeded 30 000 microbe cells. Parenteral infection with plague agent did not affect essentially the longevity of fleas.     

Antibiotic sensitivity of plague microbe strains from foreign countries]

The method of serial dilutions on the Hottinger agar was applied to comparative assay of antibiotic sensitivity in 50 strains of the plague microbe isolated abroad and in 5 strains isolated in the plague focus in the Central Caucasus. The antibiotics used in the assay were the following: streptomycin, gentamicin, doxycycline, monomycin, kanamycin, tetracycline, erythromycin, ristomycin, lincomycin and polymyxin M. Irrespective of the origin, all the isolates were resistant to erythromycin, lincomycin and polymyxin M. The levels of the sensitivity to the other antibiotics were different. The data serve as a ground for the statement that there is no tendency to development of antibiotic resistance in the plague microbe in patients treated with high doses of the antibiotics and mainly streptomycin. Along with streptomycin, such antibiotics as gentamicin, tetracycline, doxycycline and kanamycin are useful in the therapy of plague and require further investigation.     

A comparative seroepidemiologic study of the neutralizing antibody against the virulent standard strains and the Sabin vaccine strains of poliovirus among healthy Japanese

The neutralizing antibody of 160 serum specimens collected in 1978 from healthy residents in five prefectures in Japan was titrated against both the virulent standard strains and the Sabin vaccine strains of three types of poliovirus. Antibody-positive rates with both strains of respective types at a level of 1:4 were comparable in all three types of poliovirus. However, the geometric mean titers (GMTs) obtained against both strains showed statistically significant difference depending on the age-cohort's previous history of exposure to the wild or the vaccine strains of polioviruses: the younger age cohorts showed higher GMTs to the Sabin strains, while adults responded higher to the virulent standard strains. The difference was most pronounced in type 1.     

The expression of Fc gamma receptors on the surface of macrophages activated by a vaccinal strain of Yersinia pestis

The expression of Fc gamma R on the surface of macrophages in the process of antiplague immunity formation is analyzed. The stud is performed on the alveolar and peritoneal macrophages obtained from intact and immunized guinea pigs in different periods after vaccination (the 1st, 7th, 14th and 21st day). It is established that during the formation of the antiplague immunity there occurs activation of macrophages which is accompanied by an increase of the Fc gamma R expression on the outer surface of the membrane both of peritoneal and alveolar macrophages and the pattern of response of these cells to the interaction with the vaccine strain of the plague microbe changes. The Fc gamma R expression heterogeneity of certain macrophage populations is revealed both in an intact and in immune organism as well as different pattern of the intact alveolar and peritoneal macrophage response during the interaction with the vaccine strain of the antiplague microbe. These differences are levelled in the process of the antiplague immunity formation.     

Relationship of the flea Citellophilus tesquorum altaicus (Siphonaptera: Ceratophyllidae) with the plague agent

Some aspects of relationships of the flea Citellophilus tesquorum altaicus and bacterium Yersinia pestis of two strains isolated from different parts of the Tuva natural plague focus were studied. Peculiarities of elimination and blood meal activity of fleas infected with two strains of the plague agent were not revealed. Differences in mortality and alimentary activity are considerably determined by the sex of insects. The ability of examined strains to form a proventriculus block was not identical in the strains examined. This ability was expressed higher in the strain I-3428, which originated from the same part of the natural focus as the insectarium flea culture, than in the strain I-3327. During the spring and first half of summer, the proventriculus block appeared more frequently in females. The increasing of the fraction of blocked individuals was observed in both sexes from spring to summer. As for the ability to transmit the plague agent, similar seasonal increasing was noted in males, but in females, the ability to inoculate the plague microbe was always maintained at the same level.