The ability of flea species on the common vole from mountainous Dagestan to transmit and preserve the causative agent of plague

The infection ability and terms of preservation of plague microbe in fleas of common vole from Dagestan high-mountain plague focus (Frontopsylla causasica, Megabothris turbidus, Ctenophthalmus intermedius and Amphipsylla rossica) was studied experimentally. Block formation and transmission of the infection by F. caucasica and M. turbidus was observed. The transmission of plague microbe by Ct. intermedius fleas was first carried out. These species of fleas preserve the infection for a long time. Experiments with fleas of A. rossica yielded negative results.     

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.     

Epizootiological importance of Frontopsylla hetera (Siphonaptera) fleas in the Gorno-Altai natural plague focus

Experiments conducted during all seasons have established that F. hetera, one of the mass species of fleas in Mountain Altai, can be infected both by the strain of selective virulence typical to this nidus and by the non-typical non-virulent mountain-altai strain of plague agent. The non-virulent strain does not form in fleas the block of proventriculus and within 1.5-2 months they become free from the microbe. At the infection with the typical strain of the altai subspecies rare transmissions of the agent to Pallas' pika can take place as well as its long preservation in fleas.     

The efficiency of Rhadinopsylla rothschildi and R. dahurica fleas as vectors of the causative agent of plague in a Transbaikal natural focus

Experiments have shown that a block of proventriculus arises in 2.1 to 12.5% of infected fleas of R. rothschildi and in 7.2 to 10.5% of R. dahurica. These fleas transmit the plague agent to different animals (Brandt's vole, narrow-skulled vole, Dahurian suslik). The plague microbe is preserved in the organism of infected insects till the end of the experiment, 74 and 24 days respectively. Therefore, the fleas can play a part in the maintenance of plague epizootics in the Transbaikal natural nidus.     

Efficiency of Neopsylla abagaitui fleas as plague vectors in Transbaikalia

Experiments have shown that the block of proventriculus develops in 1.1 to 1.5% of individuals of the flea Neopsylla abagaitui infected with plague microbe. These insects transmit the agent during bloodsucking to different plague carriers (Citellus dauricus, C. undulatus, Meriones unguiculatus, Microtus gregalis). The plague microbe is preserved in fleas for 65 days (the observation period).     

Experimental study of the infecting ability of the flea Coptopsylla lamellifer rostrata in the Kyzylkum natural focus of plague

Experimental infecting of the fleas Coptopsylla lamellifer rostrata by plague was carried out. The fleas were infected by feeding through biomembrane (skin of a white mouse) with a mixture of defibrinated blood of guinea-pig and plague microbes. Under concentration 2-3 milliard of the microbes in 1 millilitre, from 60 to 100% of the fleas were infected. Forming of the block of proventriculus was observed in 9-13 day. Mean percent of blocking for all experiments was less than 1%. Maximal rate of blocking (2.8%) was observed in the experiment under everyday 3-hour feeding. After planting of the fleas with blocks on great gerbil (2 fleas on one gerbil) 20% of the gerbils were infected. The conclusion had been made that C. lamellifer rostrata has only insignificant effectiveness as a vector of plague agent in Kyzylkum. Probably it is caused by some specific features of this subspecies.     

The epizootiological significance of flea accumulation in aggregations of the female long-tailed suslik in the Tuva plague focus

The formation of aggregations of related females is a peculiarity of the intrapopulational organization of long-tailed suslik that results in the accumulation of Citellophilus tesquorum, the main vector and keeper of the plague microbe in the Tuva plague nidus. The plague agent is adapted to the existence on the territory occupied by aggregations of females that manifests itself in the delay of the beginning and prolixity of block-formation periods in fleas. The transfer of the agent from one place to another takes place only at the high abundance of long-tailed suslik. When the number of the suslik is low the plague microbe circulates in the ranges of one or several neighbouring aggregations of females having no possibility for successful transfer to a new place.     

Pathogenic action of the plague microbe on the flea Xenopsylla cheopis and the ultrastructure of the causative agent at various times of its stay in the vector

Pathology of gastro-intestinal tract of Xenopsylla cheopis fleas infected with plague microbe was determined by means of electron microscopy. Ultrastructure of plague microbe during different periods of its stay in the vector was studied.     

Effect of the digestive process on the survival of the plague agent in Xenopsylla gerbilli Minax fleas

Hystological investigations of experimentally infected X. gerbilli minax have shown that at the early stages of the digestion, while in the intestine compact clots of non-digested blood, are preserved, proceeds the reproduction of the plague agent. With the decay of the alimentary clot it changes into the dying off of the microbe population and after the ceasing of digestion the accumulation of bacteria renews again. Simultaneously with the changes in the number of microbes their morphology varies. At the initial stage of blood digestion bacilli typical for the agent are dominant. With the decay of the alimentary clot they are replaced by ovoid and spheroid forms. After the ceasing of the digestion cycle the microbe mass is represented by small coccobacteria. During the decrease in the plague agent abundance bacteria are absent mostly often from the midgut and posterior parts of the alimentary canal. Most favourable conditions for their preservation exist in this period in the proventriculus and in adjoining to it oesophagus part.     

The epizootiological role of the population organization of the stock of fleas on the long-tailed suslik in a natural focus of plague in Tuva

Spatial location of epizootic events in the Tuva plague focus is determined at a considerable degree by the population structure of the flea Citellophilus tesquorum--the main plague microbe vector. Within the enzootic territory occupied by five populations of the long-tailed ground squirrel (Citellus undulatus) there are six populations of C. tesquorum. Each population of fleas has a corresponding autonomic plague focus. Various conditions for the microbe life activity in these populations are recovered. The circulation of the microbe is closely connected with certain intrapopulation groupings of fleas--the nuclei of populations.     

Antagonistic and insecticidal properties of soil micromycetes

It was shown that microscopic fungi isolated in plague foci had an antagonistic effect on the plague causative agent. In the majority of the cases it was associated with their insecticidal activity against plague-transmitting fleas. It is expedient to consider the relations revealed between the burrow biocenosis components in investigation of plague enzootic aspects and development of new biological insecticides for control of the infection carriers.     

Block formation capacity in fleas infected with a plague agent after their feeding with blood substitutes

By feeding plague infected Xenopsylla cheopis fleas with blood substitutes free of blood formed elements blocked fleas were obtained. These fleas are able to transmit plague agent to white mice and cause their mortality.     

Seasonal dynamics of infecting ability of the flea Citellophilus tesquorum altaicus in the Tuva natural focus of the plague

The infecting ability of the fleas Citellophilus tesquorum altaicus loff, 1936, the main plague vectors in the Tuva natural focus, was experimentally studied in different periods of the epizootic season. Seasonal dynamics in the efficiency of infecting the long-tailed Siberian souslik with the plague causative agent through flea bites was noticed. Seasonal differences in infectivity of the "blocked" flea bites are revealed. An increase of infected experimental animals with a generalization of infection process in the period of epizooty activation in the natural focus was observed. A resistance of the long-tailed Siberian souslik to the plague agent infection through flea bites in the spring season was registered.     

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.     

Broader investigation into the external environment of the specific antigen of the infectious agent in epizootiological observation and study of the structure of natural foci of plague.

The possibilities of detection of the infectious agent in natural foci of plague can be expanded to some extent by using the method of search for the specific FI capsular antigen of the plague microbe in such objects of the external environment which have not yet been subjected to laboratory analysis and serological analysis in particular. Such objects include soil from the burrow and the substrate of the nest of the rodent as well as the fleas of rodents and regurgitations of birds of prey.The effectivity and suitability of examination of these objects using immuno-adsorption reactions with standard erythrocyte diagnostics (passive haemagglutination reaction, antibody neutralization reaction) were demonstrated experimentally and confirmed under field conditions in various geographical and ecological zones of the Caucasus. Thus in 1971, an earlier unknown natural focus of plague was found in the Mount Elbrus massif by means of serological analysis of the substrate of the nest and of the soil from the burrows of rodents. It can be expected that broadened serological search for the FI antigen of plague will prove useful in the study of the structure of natural foci of plague, in the reconnaissance or retrospective investigation of poorly explored territories and in the solution of some questions concerning the epizootiology of plague, such as the survival of the infectious agent in the inter-epizootic period, telluric plague, etc.     

Seasonal dynamics of epizootic process of the plague agent <Emphasis Type="Italic">Yersinia pestis</Emphasis> transmission to the long-tailed suslik <Emphasis Type="Italic">Citellus undulatus</Emphasis> by the flea <Emphasis Type="Italic">Citellophilus tesquorum</Emphasis> in Tuva

Transmission of Yersinia pestis to the long-tailed suslik (Citellus undulatus) by fleas (Citellophilus tesquorum) in the Tuva natural plague focus in different seasons (spring, summer, and autumn) was studied experimentally. Between feeding periods, insects were kept in an artificial nest under temperature and humidity closely corresponding to seasonal ones. The character of the agent transmission was estimated according to the fraction of fleas with the agent in the aggregated state (bacterial lumps, partial blocks of proventriculus), the fraction of blocked individuals, and the fraction of infected susliks and of those with the generalized form of infection. Seasonal dynamics of epizootic process of the Y. pestis transmission corresponded to the results obtained in the epizootic examination of the Tuva natural plague focus and reflected the dynamics of the epizootic process (increase-peak-decline). The activity of the formation of a proventriculus block in C. t. altaicus, the infection ability of the fleas, and the sensitivity of long-tailed Siberian susliks to Y. pestis were the highest in mid-summer (July-first ten days of August), during the period of epizooty activation in the focus. The maximal number of C. t. altaicus with the plague agent at the aggregated state was observed in the cold period, before wintering of insects and after their hibernation.     

Level of parasitic contact in colonies of the little suslik in the semideserts of western Kazakhstan

Field experiments were conducted during the settling of young animals of Citellus pygmaeus in four areas 4 to 13.6 hectares in size. From 3 to 6 animals were caught in each area and labeled by administering radioactive substance (0.5 micrograms of glycine 14C or methionine 35S) and colouring hair on the head. In a day or two labeled sousliks were caught again and in 6-7 days a total catching of animals and collection of ectoparasites from their hair and burrows done. In al, 942 sousliks were caught in the experimental areas and 2624 specimens of fleas of different species and 2305 ticks of R. schulzei collected. The average number of fleas which were labeled on one little souslik for a day was 0.1 to 2.3. The number of labeled fleas in the above areas coincided with the number of fleas infected with plague in the areas of epizootia. The level of parasitic contact in settlements of little souslik cannot provide the development of plague epizootia by transmission of the agent through fleas.     

Doxycycline in the prevention of experimental plague induced by plague microbe variants]

A comparative study was performed on the efficacy of doxycycline in experimental plague infection induced in albino mice by strain 231 of the plague microbe and its variant 231 Fra- deprived of the ability to produce the fraction I antigen. It was shown that the LD50 for strain 231 during animal treatment with doxycycline was significantly higher than that for variant 231 Fra-. Prophylaxis of the plague infection caused by the Fra- forms of the plague microbe required significantly higher doses of doxycycline (ED50) than that of the infection caused by the Fra+ forms. The use of the daily maximum permissible doses of doxycycline (50 to 100 mg/kg a day) for 10 days in treatment of albino mice infected with the strain Fra- did not provide animal survival at the level higher than 60 to 70 per cent while the survival rate in the animals infected with the strain Fra+ of the plague microbe and treated according to the same scheme amounted to 90-100 per cent. The lower therapeutic efficacy of doxycycline in the treatment of the infection caused by the fractionless variant of the plague microbe should be considered in development of rational schemes for prophylaxis and treatment of plague.     

The role of fleas (Siphonaptera) in the epizootiology of plague

This review concerns the role of the fleas in survival and spread of the plague, their influence on the seasonal dynamics of the epizootics, and infection rates of these insects in different natural foci. The critical evaluation is given to the data which are used to calculate the flea transmission probability for mathematical simulation of plague epizootics.     

Characterization of the rat pneumonic plague model: infection kinetics following aerosolization of Yersinia pestis CO92

Yersinia pestis, the causative agent of human bubonic and pneumonic plague, is spread during natural infection by the fleas of rodents. Historically associated with infected rat fleas, studies on the kinetics of infection in rats are surprisingly few, and these reports have focused mainly on bubonic plague. Although the natural route of primary infection results in bubonic plague in humans, it is commonly thought that aerosolized Y. pestis will be utilized during a biowarfare attack. Accordingly, based on our previous characterization of the mouse model of pneumonic plague, we sought to examine the progression of infection in rats exposed in a whole-body Madison chamber to aerosolized Y. pestis CO92. Following an 8.6 LD₅₀ dose of Y. pestis, injury was apparent in the rat tissues based on histopathology, and chemokines and cytokines rose above control levels (1 h post infection [p.i.]) in the sera and organ homogenates over a 72-h infection period. Bacteria disseminated from the lungs to peripheral organs, with the largest increases in the spleen, followed by the liver and blood at 72 h p.i. compared to the 1 h controls. Importantly, rats were as sensitive to pneumonic plague as mice, having a similar LD₅₀ dose by the intranasal and aerosolized routes. Further, we showed direct transmission of plague bacteria from infected to uninfected rats. Taken together, the data allowed us to characterize for the first time a rat pneumonic plague model following aerosolization of Y. pestis.