Plague In Mice Research Articles

The New South Wales Mouse Plague 2020-2021: A One Health description

A mouse plague occurred in Eastern Australia from spring 2020 to winter 2021, impacting an area of around 180,000 km2. It harmed human physical and psychological health, damaged the natural and built environment, and endangered farmed, domestic and native animals. However, the mouse plague was overshadowed by the COVID-19 pandemic, especially as the end of the plague coincided with the arrival and surge of the COVID-19 delta strain in rural New South Wales (NSW). In this article, we systematically overview the multiple impacts of the plague and highlight their complex interactions. Using a One Health framework, we comprehensively review the i) human, ii) animal and iii) environmental impacts including economic dimensions. Given the damage that the mouse plague caused to infrastructure, we consider the environment from two perspectives: the natural and the built environment. This One Health description of the 2020–2021 mouse plague identifies priorities for preparedness, response and recovery at local, regional land levels to inform response and management of future mouse plague events in Australia. It also highlights the need for ongoing collaboration between researchers and practitioners in the human, animal and environmental health sectors.

Effects of harvesting and stubble management on abundance of pest rodents (Mus musculus) in a conservation agriculture system.

The shift to more environmentally sensitive agricultural practices over the last several decades has changed farmland landscapes worldwide. Changes including no-till and retaining high biomass mulch has been coincident increases in rodent pests in South Africa, India, South America and Europe, indicating a possible conflict between conservation agriculture (CA) and rodent pest management. Research on effects of various crop management practices associated with conservation agriculture on pest rodent population dynamics is needed to anticipate and develop CA-relevant management strategies. During the Australian 2020-2021 mouse plague, farmers used post-harvest stubble management practices, including flattening and/or cutting, to reduce stubble cover in paddocks to lessen habitat suitability for pest house mice. We used this opportunity to assess the effects of both harvest and stubble management on the movement and abundance of mice in paddocks using mouse trapping and radio tracking. We found that most tracked mice remained resident in paddocks throughout harvest, and that mouse population abundance was generally unaffected by stubble management. Recent conversions to conservation agriculture practices have changed how pest house mice use cropped land. Management practices that reduce post-harvest habitat complexity do not appear to reduce the attractiveness of paddocks to mice, and further research into new management strategies in addition to toxic bait use is required as part of an integrated pest management approach. This article is protected by copyright. All rights reserved.

Lung Resident Memory T Cells Activated by Oral Vaccination Afford Comprehensive Protection against Pneumonic Yersinia pestis Infection.

A growing body of evidence has shown that resident memory T (TRM) cells formed in tissue after mucosal infection or vaccination are crucial for counteracting reinfection by pathogens. However, whether lung TRM cells activated by oral immunization with Yptb1(pYA5199) play a protective role against pneumonic plague remains unclear. In this study, we demonstrated that lung CD4+ and CD8+ TRM cells significantly accumulated in the lungs of orally Yptb1(pYA5199)-vaccinated mice and dramatically expanded with elevated IL-17A, IFN-γ, and/or TNF-α production after pulmonary Yersinia pestis infection and afforded significant protection. Short-term or long-term treatment of immunized mice with FTY720 did not affect lung TRM cell formation and expansion or protection against pneumonic plague. Moreover, the intratracheal transfer of both lung CD4+ and CD8+ TRM cells conferred comprehensive protection against pneumonic plague in naive recipient mice. Lung TRM cell-mediated protection was dramatically abolished by the neutralization of both IFN-γ and IL-17A. Our findings reveal that lung TRM cells can be activated via oral Yptb1(pYA5199) vaccination, and that IL-17A and IFN-γ production play an essential role in adaptive immunity against pulmonary Y. pestis infection. This study highlights an important new target for developing an effective pneumonic plague vaccine.

Lymphocytic Choriomeningitis Virus Infection, Australia.

During a mouse plague in early 2021, a farmer from New South Wales, Australia, sought treatment for aseptic meningitis and was subsequently diagnosed with locally acquired lymphocytic choriomeningitis virus infection. Whole-genome sequencing identified a divergent and geographically distinct lymphocytic choriomeningitis virus strain compared with other published sequences.

Acute oral toxicity of zinc phosphide: an assessment for wild house mice (Mus musculus).

Irregular plagues of house mice, Mus musculus, incur major economic impacts on agricultural production in Australia. The efficacy of zinc phosphide (ZnP), the only registered broadacre control agent for mice, is reported as increasingly variable. Have mice become less sensitive over time or are they taking a sub-lethal dose and developing aversion? In this laboratory study, the sensitivity of mice (wild caught; outbred laboratory strain) was assessed using oral gavage of a range of ZnP concentrations. The estimated LD50 values (72-79 mg ZnP/kg body weight) were similar for each mouse group but are significantly higher than previously reported. The willingness of mice to consume ZnP-coated grains was determined. ZnP-coated grains (50 g ZnP/kg grain) presented in the absence of alternative food were consumed and 94% of wild mice died. Mice provided with alternative food and ZnP-coated wheat grains (either 25 or 50 g ZnP/kg grain) consumed toxic and non-toxic grains, and mortality was lower (33-55%). If a sublethal amount of ZnP-coated grain was consumed, aversion occurred, mostly when alternative food was present. The sensitivity of wild house mice to ZnP in Australia is significantly lower than previously assumed. Under laboratory conditions, ZnP-coated grains coated with a new higher dose (50 g ZnP/kg grain) were readily consumed. Consumption of toxic grain occurred when alternative food was available but was decreased. Our unambiguous findings for house mice indicate a re-assessment of the ZnP loading for baits used for control of many rodents around the world may be warranted.

Simulation of Bubonic Plague in BSL-2 Laboratory

The causative agent of plague, Yersinia pestis, is classified as pathogenicity (hazard) group I agent, which means that the work with “wild type” strains should be carried out in BSL-3 facilities. Y. pestis EV NIIEG is a Δpgm strain, allowing experimental studies to be carried out in BSL-2 laboratories. However, the disease and its progression elicited by such strain do not entirely mirror the infection observed with fully virulent strains. Residual virulence of Y. pestis EV NIIEG strain for mice can be increased under in vivo iron supplementation. The aim of the study was to optimize methodological approaches to modeling experimental plague in laboratory animals following administration of attenuated Δpgm Y. pestis strains with iron dextran. Materials and methods. Simulation of plague infection in outbred mice was carried out through subcutaneous inoculation of Y. pestis EV NIIEG strain with iron dextran supplementation. The animal condition was assessed on a daily basis. In the course of the experiment, the pathological presentation and bacterial content in organs of mice were evaluated. Results and discussion. Mice inoculated subcutaneously with Y. pestis EV NIIEG strain in the presence of iron dextran developed a bubonic plague that resulted in lethal outcome with pathological changes of internal organs, characteristic of plague infection. In case of daily administration of iron, LD50 of Y. pestis EV NNIEG strain for the mice significantly exceeded the same one with a single injection. Differences in the survival rate among animals in the groups with a single and multiple administration of iron compared to the control group were statistically valid. Thus, attenuated Δpgm Y. pestis strains in the presence of iron dextran can be used to model experimental plague in mice with marked pathological changes and lethality in BSL-2 laboratories.

Impacts of House Mice on Sustainable Fodder Storage in Australia

Mice cause substantial economic, social, and environmental damage to rural production systems and communities in Australia, especially during mouse plagues. The losses and damage caused by mice to hay/fodder storage are not well understood, given the size of the industry, so a pilot study (survey of 14 growers) was designed to better understand the physical and economic damage, consider disease implications, and identify the management strategies available. During a mouse plague, mice were regarded as the only factor (other than weather) that affected the long-term sustainability of fodder storage. Growers were feeding fodder to livestock (sheep/cattle) at twice the rate they normally would because of mouse damage and contamination. Mice damaged strings and the structure of bales, making transport impractical, leading to destruction of some stacks. Losses caused by mice were estimated at AUD 140,000 (range = AUD 7000–461,580; equivalent to 30–40% loss of value) and included estimates of physical damage to hay/fodder bales, rodenticides, and labour costs. Growers were concerned about contamination and disease transfer to livestock (and workers) from mice through urine/faeces and their carcasses, especially during mouse plagues. There are significant gaps in our knowledge on the impacts of mice to fodder storage in Australia. Research is needed to (1) identify effective mouse control options, (2) determine the economic impact of mice, and (3) undertake a disease study of mice, to help inform appropriate management strategies for effective control.

A most enigmatic mouse: additional information on collection of blue-grey mouse (

Australia has suffered more modern mammal extinctions than any nation. Among the list of extinct species is the blue-grey mouse (Pseudomys glaucus), a rodent known from just three specimens (two from southern Queensland and one from northern New South Wales). We investigated circumstances of collection for the three specimens referred to this species to better illuminate optimal habitats and climatic conditions for continuing rediscovery efforts. No additional information could be found on two specimens from southern Queensland sent to the Natural History Museum, London. However, we recorded a first-hand account of how the northern New South Wales specimen was collected and have established that the collection year was 1956. We also obtained a copy of correspondence from Ellis Le G. Troughton thanking the collector for the contribution. The northern New South Wales specimen was from a pastoral property formerly dominated by woodland vegetation communities on alluvial soils. It was captured during a mouse plague following consecutive seasons of above average rainfall. Pseudomys glaucus is likely already extinct, but our results help better direct any future survey efforts. Surveys should be targeted in woodland communities on alluvial floodplains in the Darling Riverine Plains bioregion, following periods of above average rainfall conducive to the irruption of rodent populations.

It’s a trap: effective methods for monitoring house mouse populations in grain-growing regions of south-eastern Australia

Abstract Context Wild house mice cause substantial economic damage to grain crops in Australia, particularly during mouse plagues. Populations were monitored to detect changes in abundance, with data from surveys used in models to forecast likely mouse outbreaks. However, it is not always feasible to use live-trapping (the ‘gold standard’) for assessing mouse abundance at a large number of monitoring sites spread across south-eastern Australia. A range of alternative methods was tried to assist the grains industry with strategic decisions to reduce crop damage. Aims The aim of this work was to determine which survey methods could provide useful and effective indexes of mouse abundance across a large area. Methods Monitoring of mouse populations was conducted at representative grain farms by using (1) live-trapping at long-term ‘benchmark’ sites (n = 2), and (2) mouse chew cards and active burrow counts at ‘rapid-assessment’ sites (n = 44 farms across 5 regions). Monitoring was conducted for 22 monitoring sessions over 7.5 years through low, medium and high mouse abundance conditions. Key results Live-trapping provided the most useful, but most resource-intensive, information. There were strong relationships between the index of mouse abundance from live-trapping with mouse chew cards and active burrow counts at a local (explaining 63% and 71% of variation respectively) and regional (explaining 71% and 81% of variation respectively) scales. The same quantitative relationship held between the mouse chew cards and trapping regardless of season and year. However, the relationship between active burrow counts and trapping was best in winter and autumn seasons. There was a strong relationship between mouse abundance from live-trapping and active burrows across 1 ha grids (R2 = 0.88). We determined there were 1.3 ± 0.2 (mean ± s.e.) mice per active burrow. Conclusions Live-trapping supplemented with data from chew cards and active burrows remains sufficient to monitor a wide range of sites to show regional trends. Implications It is likely that live-trapping will need to be used for the foreseeable future to provide useful parameters such as breeding condition and population abundance that are required for the forecast models. Supplementary monitoring at rapid-assessment sites (using chew cards in all seasons and active burrow counts particularly in autumn and winter), that can be collected easily without the need for animal handling, will provide additional indications of region-specific changes in mouse abundance and activity.

Effects of background food on alternative grain uptake and zinc phosphide efficacy in wild house mice.

BACKGROUNDHouse mice (Mus musculus) cause significant, ongoing losses to grain crops in Australia, particularly during mouse plagues. Zinc phosphide (ZnP) coated grain is used for control, but with variable success. In a laboratory setting, we tested if mice would (i) switch from consumption of one grain type to another when presented with an alternative and (ii) consume ZnP‐treated grains when presented as a choice with a different grain.RESULTSMice readily switched from their background grain to an alternative grain, preferring cereals (wheat or barley) over lentils. Mice readily consumed ZnP‐coated barley grains. Their mortality rate was significantly higher (86%, n = 30) in the presence of a less‐favoured grain (lentils) compared to their mortality rate (47%, n = 29; 53%, n = 30) in the presence of a more‐favoured grain (wheat and barley, respectively). Mice died between 4 and 112 h (median = 18 h) after consuming one or more toxic grains. Independent analysis of ZnP‐coated grains showed variable toxin loading indicating that consumption of a single grain would not guarantee intake of a lethal dose. There was also a strong and rapid behavioural aversion if mice did not consume a lethal dose on the first night.CONCLUSIONSThe registered dose rate of 25 g of ZnP/kg wheat (~1 mg of ZnP/grain) in Australia needs to be re‐evaluated to determine what factors may be contributing to variation in efficacy. Further field research is also required to understand the complex association between ZnP dose, and quantity and quality of background food on efficacy of ZnP baits.

Multiple ecological processes underpin the eruptive dynamics of small mammals: House mice in a semi-arid agricultural environment.

Mouse plagues are a regular feature of grain‐growing regions, particularly in southern and eastern Australia, yet it is not clear what role various ecological processes play in the eruptive dynamics generating these outbreaks.This research was designed to assess the impact of adding food, water, and cover in all combinations on breeding performance, abundance, and survival of mouse populations on a typical cereal growing farm in northwestern Victoria.Supplementary food, water, and cover were applied in a 2 × 2 × 2 factorial design to 240 m sections of internal fence lines between wheat or barley crops and stubble/pasture fields over an 11‐month period to assess the impact on mouse populations.We confirmed that mice were eating the additional food and were accessing the water provided. We did not generate an outbreak of mice, but there were some significant effects from the experimental treatments. Additional food increased population size twofold and improved apparent survival. Both water and cover improved breeding performance. Food and cover increased apparent survival.Our findings confirm that access to food, water, and cover are necessary for outbreaks, but are not sufficient. There remain additional factors that are important in generating mouse plagues, particularly in a climatically variable agricultural environment.

A Recombinant Attenuated Yersinia pseudotuberculosis Vaccine Delivering a Y. pestis YopENt138-LcrV Fusion Elicits Broad Protection against Plague and Yersiniosis in Mice.

In this study, a novel recombinant attenuated Yersinia pseudotuberculosis PB1+ strain (χ10069) engineered with ΔyopK ΔyopJ Δasd triple mutations was used to deliver a Y. pestis fusion protein, YopE amino acid 1 to 138-LcrV (YopENt138-LcrV), to Swiss Webster mice as a protective antigen against infections by yersiniae. χ10069 bacteria harboring the pYA5199 plasmid constitutively synthesized the YopENt138-LcrV fusion protein and secreted it via the type 3 secretion system (T3SS) at 37°C under calcium-deprived conditions. The attenuated strain χ10069(pYA5199) was manifested by the establishment of controlled infection in different tissues without developing conspicuous signs of disease in histopathological analysis of microtome sections. A single-dose oral immunization of χ10069(pYA5199) induced strong serum antibody titers (log10 mean value, 4.2), secretory IgA in bronchoalveolar lavage (BAL) fluid from immunized mice, and Yersinia-specific CD4+ and CD8+ T cells producing high levels of tumor necrosis factor alpha (TNF-α), gamma interferon (IFN-γ), and interleukin 2 (IL-2), as well as IL-17, in both lungs and spleens of immunized mice, conferring comprehensive Th1- and Th2-mediated immune responses and protection against bubonic and pneumonic plague challenges, with 80% and 90% survival, respectively. Mice immunized with χ10069(pYA5199) also exhibited complete protection against lethal oral infections by Yersinia enterocolitica WA and Y. pseudotuberculosis PB1+. These findings indicated that χ10069(pYA5199) as an oral vaccine induces protective immunity to prevent bubonic and pneumonic plague, as well as yersiniosis, in mice and would be a promising oral vaccine candidate for protection against plague and yersiniosis for human and veterinary applications.

Combating Multidrug-Resistant Pathogens with Host-Directed Nonantibiotic Therapeutics.

Earlier, we reported that three Food and Drug Administration-approved drugs, trifluoperazine (TFP; an antipsychotic), amoxapine (AXPN; an antidepressant), and doxapram (DXP; a breathing stimulant), identified from an in vitro murine macrophage cytotoxicity screen, provided mice with 40 to 60% protection against pneumonic plague when administered at the time of infection for 1 to 3 days. In the present study, the therapeutic potential of these drugs against pneumonic plague in mice was further evaluated when they were administered at up to 48 h postinfection. While the efficacy of TFP was somewhat diminished as treatment was delayed to 24 h, the protection of mice with AXPN and DXP increased as treatment was progressively delayed to 24 h. At 48 h postinfection, these drugs provided the animals with significant protection (up to 100%) against challenge with the agent of pneumonic or bubonic plague when they were administered in combination with levofloxacin. Likewise, when they were used in combination with vancomycin, all three drugs provided mice with 80 to 100% protection from fatal oral Clostridium difficile infection when they were administered at 24 h postinfection. Furthermore, AXPN provided 40 to 60% protection against respiratory infection with Klebsiella pneumoniae when it was administered at the time of infection or at 24 h postinfection. Using the same in vitro cytotoxicity assay, we identified an additional 76/780 nonantibiotic drugs effective against K. pneumoniae For Acinetobacter baumannii, 121 nonantibiotic drugs were identified to inhibit bacterium-induced cytotoxicity in murine macrophages. Of these 121 drugs, 13 inhibited the macrophage cytotoxicity induced by two additional multiple-antibiotic-resistant strains. Six of these drugs decreased the intracellular survival of all three A. baumannii strains in macrophages. These results provided further evidence of the broad applicability and utilization of drug repurposing screening to identify new therapeutics to combat multidrug-resistant pathogens of public health concern.

Zinc transporters YbtX and ZnuABC are required for the virulence of Yersinia pestis in bubonic and pneumonic plague in mice.

A number of bacterial pathogens require the ZnuABC Zinc (Zn2+) transporter and/or a second Zn2+ transport system to overcome Zn2+ sequestration by mammalian hosts. Previously we have shown that in addition to ZnuABC, Yersinia pestis possesses a second Zn2+ transporter that involves components of the yersiniabactin (Ybt), siderophore-dependent iron transport system. Synthesis of the Ybt siderophore and YbtX, a member of the major facilitator superfamily, are both critical components of the second Zn2+ transport system. Here we demonstrate that a ybtX znu double mutant is essentially avirulent in mouse models of bubonic and pneumonic plague while a ybtX mutant retains high virulence in both plague models. While sequestration of host Zn is a key nutritional immunity factor, excess Zn appears to have a significant antimicrobial role in controlling intracellular bacterial survival. Here, we demonstrate that ZntA, a Zn2+ exporter, plays a role in resistance to Zn toxicity in vitro, but that a zntA zur double mutant retains high virulence in both pneumonic and bubonic plague models and survival in macrophages. We also confirm that Ybt does not directly bind Zn2+in vitro under the conditions tested. However, we detect a significant increase in Zn2+-binding ability of filtered supernatants from a Ybt+ strain compared to those from a strain unable to produce the siderophore, supporting our previously published data that Ybt biosynthetic genes are involved in the production of a secreted Zn-binding molecule (zincophore). Our data suggest that Ybt or a modified Ybt participate in or promote Zn-binding activity in culture supernatants and is involved in Zn acquisition in Y. pestis.

A Replication-Defective Human Type 5 Adenovirus-Based Trivalent Vaccine Confers Complete Protection against Plague in Mice and Nonhuman Primates.

Currently, no plague vaccine exists in the United States for human use. The capsular antigen (Caf1 or F1) and two type 3 secretion system (T3SS) components, the low-calcium-response V antigen (LcrV) and the needle protein YscF, represent protective antigens of Yersinia pestis We used a replication-defective human type 5 adenovirus (Ad5) vector and constructed recombinant monovalent and trivalent vaccines (rAd5-LcrV and rAd5-YFV) that expressed either the codon-optimized lcrV or the fusion gene designated YFV (consisting of ycsF, caf1, and lcrV). Immunization of mice with the trivalent rAd5-YFV vaccine by either the intramuscular (i.m.) or the intranasal (i.n.) route provided protection superior to that with the monovalent rAd5-LcrV vaccine against bubonic and pneumonic plague when animals were challenged with Y. pestis CO92. Preexisting adenoviral immunity did not diminish the protective response, and the protection was always higher when mice were administered one i.n. dose of the trivalent vaccine (priming) followed by a single i.m. booster dose of the purified YFV antigen. Immunization of cynomolgus macaques with the trivalent rAd5-YFV vaccine by the prime-boost strategy provided 100% protection against a stringent aerosol challenge dose of CO92 to animals that had preexisting adenoviral immunity. The vaccinated and challenged macaques had no signs of disease, and the invading pathogen rapidly cleared with no histopathological lesions. This is the first report showing the efficacy of an adenovirus-vectored trivalent vaccine against pneumonic plague in mouse and nonhuman primate (NHP) models.

A Recombinant Raccoon Poxvirus Vaccine Expressing both Yersinia pestis F1 and Truncated V Antigens Protects Animals against Lethal Plague.

In previous studies, we demonstrated in mice and prairie dogs that simultaneous administration of two recombinant raccoon poxviruses (rRCN) expressing Yersinia pestis antigens (F1 and V307—a truncated version of the V protein) provided superior protection against plague challenge compared to individual single antigen constructs. To reduce costs of vaccine production and facilitate implementation of a sylvatic plague vaccine (SPV) control program for prairie dogs, a dual antigen construct is more desirable. Here we report the construction and characterization of a novel RCN-vectored vaccine that simultaneously expresses both F1 and V307 antigens. This dual antigen vaccine provided similar levels of protection against plague in both mice and prairie dogs as compared to simultaneous administration of the two single antigen constructs and was also shown to protect mice against an F1 negative strain of Y. pestis. The equivalent safety, immunogenicity and efficacy profile of the dual RCN-F1/V307 construct warrants further evaluation in field efficacy studies in sylvatic plague endemic areas.

Population dynamics of house mice in Queensland grain-growing areas

Context Irregular plagues of house mice cause high production losses in grain crops in Australia. If plagues can be forecast through broad-scale monitoring or model-based prediction, then mice can be proactively controlled by poison baiting. Aims To predict mouse plagues in grain crops in Queensland and assess the value of broad-scale monitoring. Methods Regular trapping of mice at the same sites on the Darling Downs in southern Queensland has been undertaken since 1974. This provides an index of abundance over time that can be related to rainfall, crop yield, winter temperature and past mouse abundance. Other sites have been trapped over a shorter time period elsewhere on the Darling Downs and in central Queensland, allowing a comparison of mouse population dynamics and cross-validation of models predicting mouse abundance. Key results On the regularly trapped 32-km transect on the Darling Downs, damaging mouse densities occur in 50% of years and a plague in 25% of years, with no detectable increase in mean monthly mouse abundance over the past 35 years. High mouse abundance on this transect is not consistently matched by high abundance in the broader area. Annual maximum mouse abundance in autumn–winter can be predicted (R2 = 57%) from spring mouse abundance and autumn–winter rainfall in the previous year. In central Queensland, mouse dynamics contrast with those on the Darling Downs and lack the distinct annual cycle, with peak abundance occurring in any month outside early spring. On average, damaging mouse densities occur in 1 in 3 years and a plague occurs in 1 in 7 years. The dynamics of mouse populations on two transects ~70 km apart were rarely synchronous. Autumn–winter rainfall can indicate mouse abundance in some seasons (R2 = ~52%). Conclusion Early warning of mouse plague formation in Queensland grain crops from regional models should trigger farm-based monitoring. This can be incorporated with rainfall into a simple model predicting future abundance that will determine any need for mouse control. Implications A model-based warning of a possible mouse plague can highlight the need for local monitoring of mouse activity, which in turn could trigger poison baiting to prevent further mouse build-up.

A Yersinia pestisYscN ATPase mutant functions as a live attenuated vaccine against bubonic plague in mice

Yersinia pestis is the causative agent responsible for bubonic and pneumonic plague. The bacterium uses the pLcr plasmid-encoded type III secretion system to deliver virulence factors into host cells. Delivery requires ATP hydrolysis by the YscN ATPase encoded by the yscN gene also on pLcr. A yscN mutant was constructed in the fully virulent CO92 strain containing a nonpolar, in-frame internal deletion within the gene. We demonstrate that CO92 with a yscN mutation was not able to secrete the LcrV protein (V-Antigen) and attenuated in a subcutaneous model of plague demonstrating that the YscN ATPase was essential for virulence. However, if the yscN mutant was complemented with a functional yscN gene in trans, virulence was restored. To evaluate the mutant as a live vaccine, Swiss-Webster mice were vaccinated twice with the ΔyscN mutant at varying doses and were protected against bubonic plague in a dose-dependent manner. Antibodies to F1 capsule but not to LcrV were detected in sera from the vaccinated mice. These preliminary results suggest a proof-of-concept for an attenuated, genetically engineered, live vaccine effective against bubonic plague.

Early Systemic Bacterial Dissemination and a Rapid Innate Immune Response Characterize Genetic Resistance to Plague of SEG Mice

Although laboratory mice are usually highly susceptible to Yersinia pestis, we recently identified a mouse strain (SEG) that exhibited an exceptional capacity to resist bubonic plague and used it to identify immune mechanisms associated with resistance. The kinetics of infection, circulating blood cells, granulopoiesis, lesions, and cellular populations in the spleen, and cytokine production in various tissues were compared in SEG and susceptible C57BL/6J mice after subcutaneous infection with the virulent Y. pestis CO92. Bacterial invasion occurred early (day 2) but was transient in SEG/Pas mice, whereas in C57BL/6J mice it was delayed but continuous until death. The bacterial load in all organs significantly correlated with the production of 5 cytokines (granulocyte colony-stimulating factor, keratinocyte-derived chemokine (KC), macrophage cationic peptide-1 (MCP-1), interleukin 1α, and interleukin 6) involved in monocyte and neutrophil recruitment. Indeed, higher proportions of these 2 cell types in blood and massive recruitment of F4/80(+)CD11b(-) macrophages in the spleen were observed in SEG/Pas mice at an early time point (day 2). Later times after infection (day 4) were characterized in C57BL/6J mice by destructive lesions of the spleen and impaired granulopoiesis. A fast and efficient Y. pestis dissemination in SEG mice may be critical for the triggering of an early and effective innate immune response necessary for surviving plague.

Yersinia pestisacrAB-tolCin Antibiotic Resistance and Virulence

The efflux pump AcrAB is important in the antibiotic resistance and virulence of several pathogenic bacteria. We report that deletion of the Yersinia pestis AcrAB-TolC homolog leads to increased susceptibility to diverse substrates, including, though unlike in Escherichia coli, the aminoglycosides. Neither is the Y. pestis pump affected by the efflux pump inhibitor phenylalanine-arginine beta-naphthylamide. In mouse plague models, pump deletion does not have a significant effect on tissue colonization.