RESUMEN
Plague, a primarily flea-borne disease caused by Yersinia pestis, is characterized by rapidly spreading epizootics separated by periods of quiescence. Little is known about how and where Y. pestis persists between epizootics. It is commonly proposed, however, that Y pestis is maintained during interepizootic periods in enzootic cycles involving flea vectors and relatively resistant host populations. According to this model, while susceptible individuals serve as infectious sources for feeding fleas and subsequently die of infection, resistant hosts survive infection, develop antibodies to the plague bacterium, and continue to provide bloodmeals to infected fleas. For Y. pestis to persist under this scenario, fleas must remain infected after feeding on hosts carrying antibodies to Y. pestis. Studies of other vector-borne pathogens suggest that host immunity may negatively impact pathogen survival in the vector. Here, we report infection rates and bacterial loads for fleas (both Xenopsylla cheopis (Rothschild) and Oropsylla montana (Baker)) that consumed an infectious bloodmeal and subsequently fed on an immunized or age-matched naive mouse. We demonstrate that neither the proportion of infected fleas nor the bacterial loads in infected fleas were significantly lower within 3 d of feeding on immunized versus naive mice. Our findings thus provide support for one assumption underlying the enzootic host model of interepizootic maintenance of Y. pestis.
Asunto(s)
Siphonaptera/inmunología , Siphonaptera/microbiología , Yersinia pestis/fisiología , Animales , Carga Bacteriana , Sangre , Conducta Alimentaria , Interacciones Huésped-Patógeno , RatonesRESUMEN
Sharp declines in human and animal cases of plague, caused by the bacterium Yersinia pestis (Yersin), have been observed when outbreaks coincide with hot weather. Failure of biofilm production, or blockage, to occur in the flea, as temperatures reach 30 degrees C has been suggested as an explanation for these declines. Recent work demonstrating efficient flea transmission during the first few days after fleas have taken an infectious blood meal, in the absence of blockage (e.g., early-phase transmission), however, has called this hypothesis into question. To explore the potential effects of temperature on early-phase transmission, we infected colony-reared Xenopsylla cheopis (Rothchild) fleas with a wild-type strain of plague bacteria using an artificial feeding system, and held groups of fleas at 10, 23, 27, and 30 degrees C. Naive Swiss Webster mice were exposed to fleas from each of these temperatures on days 1-4 postinfection, and monitored for signs of infection for 21 d. Temperature did not significantly influence the rates of transmission observed for fleas held at 23, 27, and 30 degrees C. Estimated per flea transmission efficiencies for these higher temperatures ranged from 2.32 to 4.96% (95% confidence interval [CI]: 0.96-8.74). In contrast, no transmission was observed in mice challenged by fleas held at 10 degrees C (per flea transmission efficiency estimates, 0-1.68%). These results suggest that declines in human and animal cases during hot weather are not related to changes in the abilities of X. cheopis fleas to transmit Y. pestis infections during the early-phase period. By contrast, transmission may be delayed or inhibited at low temperatures, indicating that epizootic spread of Y. pestis by X. cheopis via early-phase transmission is unlikely during colder periods of the year.
Asunto(s)
Peste/transmisión , Xenopsylla/microbiología , Yersinia pestis/fisiología , Animales , Conducta Alimentaria/fisiología , Ratones , Peste/microbiología , Temperatura , Xenopsylla/fisiologíaRESUMEN
BACKGROUND: The Centers for Disease Control and Prevention (CDC) and others reported that methicillin-resistant S. aureus (MRSA) are significant causes of serious human infections, including pulmonary illnesses. We investigated the role played by superantigens in lung-associated lethal illness in rabbits. METHODS: A rabbit model was established to investigate the potential role played by superantigens, staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), and toxic shock syndrome toxin-1 (TSST-1). Rabbits received intrabronchial community-associated (CA) MRSA strains USA200 (TSST-1(+)), MW2 (SEC(+)), c99-529 (SEB(+)), or purified superantigens. Some rabbits were preimmunized against superantigens or treated with soluble high-affinity T cell receptors (Vß-TCR) to neutralize SEB and then challenged intrabronchially with CA-MRSA or superantigens. RESULTS: Rabbits challenged with CA-MRSA or superantigens developed fatal, pulmonary illnesses. Animals preimmunized against purified superantigens, or treated passively with Vß-TCRs and then challenged with CA-MRSA or superantigens, survived. Lung histological analysis indicated that nonimmune animals developed lesions consistent with necrotizing pneumonia after challenge with CA-MRSA or purified superantigens. Superantigen-immune animals or animals treated with soluble Vß-TCRs did not develop pulmonary lesions. CONCLUSIONS: Superantigens contribute to lethal pulmonary illnesses due to CA-MRSA; preexisting immunity to superantigens prevents lethality. Administration of high-affinity Vß-TCR with specificity for SEB to nonimmune animals protects from lethal pulmonary illness resulting from SEB(+) CA-MRSA and SEB.
Asunto(s)
Enfermedades Pulmonares/inmunología , Infecciones Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Superantígenos/inmunología , Animales , Toxinas Bacterianas/administración & dosificación , Toxinas Bacterianas/inmunología , Modelos Animales de Enfermedad , Enterotoxinas/administración & dosificación , Enterotoxinas/inmunología , Enfermedades Pulmonares/microbiología , Enfermedades Pulmonares/mortalidad , Enfermedades Pulmonares/patología , Staphylococcus aureus Resistente a Meticilina/inmunología , Conejos , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/mortalidad , Infecciones Estafilocócicas/patología , Superantígenos/administración & dosificaciónRESUMEN
Early-phase transmission (EPT) is a recently described model of plague transmission that explains the rapid spread of disease from flea to mammal host during an epizootic. Unlike the traditional blockage-dependent model of plague transmission, EPT can occur when a flea takes its first blood meal after initially becoming infected by feeding on a bacteraemic host. Blockage of the flea gut results from biofilm formation in the proventriculus, mediated by the gene products found in the haemin storage (hms) locus of the Yersinia pestis chromosome. Although biofilms are required for blockage-dependent transmission, the role of biofilms in EPT has yet to be determined. An artificial feeding system was used to feed Xenopsylla cheopis and Oropsylla montana rat blood spiked with the parental Y. pestis strain KIM5(pCD1)+, two different biofilm-deficient mutants (Delta hmsT, Delta hmsR), or a biofilm-overproducer mutant (Delta hmsP). Infected fleas were then allowed to feed on naïve Swiss Webster mice for 1-4 days after infection, and the mice were monitored for signs of infection. We also determined the bacterial loads of each flea that fed upon naïve mice. Biofilm-defective mutants transmitted from X. cheopis and O. montana as efficiently as the parent strain, whereas the EPT efficiency of fleas fed the biofilm-overproducing strain was significantly less than that of fleas fed either the parent or a biofilm-deficient strain. Fleas infected with a biofilm-deficient strain harboured lower bacterial loads 4 days post-infection than fleas infected with the parent strain. Thus, defects in biofilm formation did not prevent flea-borne transmission of Y. pestis in our EPT model, although biofilm overproduction inhibited efficient EPT. Our results also indicate, however, that biofilms may play a role in infection persistence in the flea.
Asunto(s)
Biopelículas , Peste/transmisión , Yersinia pestis/fisiología , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Humanos , Insectos Vectores/microbiología , Ratones , Peste/microbiología , Ratas , Ratas Sprague-Dawley , Siphonaptera/microbiología , Yersinia pestis/genéticaRESUMEN
Most diagnostic testing for West Nile virus (WNV) disease is accomplished using serologic testing, which is subject to cross-reactivity, may require cumbersome confirmatory testing, and may fail to detect infection in specimens collected early in the course of illness. The objective of this project was to determine whether a combination of molecular and serologic testing would increase detection of WNV disease cases in acute serum samples. A total of 380 serum specimens collected ≤7 days after onset of symptoms and submitted to four state public health laboratories for WNV diagnostic testing in 2014 and 2015 were tested. WNV immunoglobulin M (IgM) antibody and RT-PCR tests were performed on specimens collected ≤3 days after symptom onset. WNV IgM antibody testing was performed on specimens collected 4-7 days after onset and RT-PCR was performed on IgM-positive specimens. A patient was considered to have laboratory evidence of WNV infection if they had detectable WNV IgM antibodies or WNV RNA in the submitted serum specimen. Of specimens collected ≤3 days after symptom onset, 19/158 (12%) had laboratory evidence of WNV infection, including 16 positive for only WNV IgM antibodies, 1 positive for only WNV RNA, and 2 positive for both. Of specimens collected 4-7 days after onset, 21/222 (9%) were positive for WNV IgM antibodies; none had detectable WNV RNA. These findings suggest that routinely performing WNV RT-PCR on acute serum specimens submitted for WNV diagnostic testing is unlikely to identify a substantial number of additional cases beyond IgM antibody testing alone.
Asunto(s)
Fiebre del Nilo Occidental/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Antivirales/sangre , Niño , Preescolar , Femenino , Humanos , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Adulto JovenRESUMEN
Yersinia pestis, the etiological agent of plague, is transmitted by multiple flea species. Previous studies have reported wide variability in transmission efficiency among competent vectors. However, it is unclear to what extent such variation is explained by methodological differences among studies. To optimize an artificial feeding system where fleas are infected with controlled numbers of Y. pestis under standardized laboratory conditions that could be used to systematically compare vector efficiency, we sought to test the effect of host bloodmeal source on (1) the flea's ability to remain infected with Y. pestis and (2) bacterial loads in fleas. Here, we demonstrate that both prevalence of infection with a virulent strain of Y. pestis (CO96-3188) and bacterial loads in rock squirrel fleas (Oropsylla montana) are affected by host-associated blood factors. The generality of this observation was confirmed by repeating the study using the rat flea (Xenopsylla cheopis) and a commonly used avirulent laboratory strain of Y. pestis (A1122). Implications of the results for rate of spread of Y. pestis in naturally infected host populations are discussed.
Asunto(s)
Sangre/microbiología , Siphonaptera/microbiología , Yersinia pestis/fisiología , Animales , Conducta Alimentaria , Ratones , Conejos , RatasRESUMEN
The role of deer mice and other species of Peromyscus as enzootic reservoirs for plague remains controversial. In this study, we evaluated early-phase vector efficiency of Aetheca wagneri Baker, a common flea species infesting deer mice, to determine the likelihood that Y. pestis could be spread mouse to mouse by this species. We showed that A. wagneri could transmit plague bacteria to laboratory mice as early as 3 d postinfection (p.i.), but transmission efficiency was quite low (1.03%; 95% CI: 0.19-3.34%) 1-4 d p.i. compared with that for the established plague vector Oropsylla montana Baker (10.63%; 95% CI: 4.18-25.91). Using this early-phase transmission efficiency estimate, we determined through parameterization of a simple predictive model that at least 68 A. wagneri per deer mouse would be required to support levels of transmission adequate for enzootic maintenance. Because deer mice typically harbor fewer than three A. wagneri per host, our data do not support the notion of an independent deer mouse--A. wagneri transmission cycle.
Asunto(s)
Interacciones Huésped-Patógeno , Peromyscus/microbiología , Peste/transmisión , Siphonaptera/microbiología , Yersinia pestis/fisiología , Animales , Infestaciones Ectoparasitarias/epidemiología , Infestaciones Ectoparasitarias/parasitología , Infestaciones Ectoparasitarias/veterinaria , Modelos Biológicos , Peromyscus/parasitología , Peste/epidemiología , Peste/veterinaria , Enfermedades de los Roedores/epidemiología , Enfermedades de los Roedores/microbiología , Enfermedades de los Roedores/parasitologíaRESUMEN
Laboratory-reared Oropsylla montana were exposed to soil and wild-caught Oropsylla montana feces for 1 week. Fleas from these two treatments and a control group of laboratory-reared fleas were infected with Yersinia pestis, the etiological agent of plague. Fleas exposed to soil transmitted Y. pestis to mice at a significantly greater rate (50.0% of mice were infected) than control fleas (23.3% of mice were infected). Although the concentration of Y. pestis in fleas did not differ among treatments, the minimum transmission efficiency of fleas from the soil and wild flea feces treatments (6.9% and 7.6%, respectively) were more than three times higher than in control fleas (2.2%). Our results suggest that exposing laboratory-reared fleas to diverse microbes alters transmission of Y. pestis.
Asunto(s)
Heces/microbiología , Siphonaptera/microbiología , Yersinia pestis/aislamiento & purificación , Animales , Ratones , Peste/microbiología , Peste/transmisión , SueloRESUMEN
We collected Oropsylla montana from rock squirrels, Spermophilus varigatus, and infected a subset of collected fleas with Yersinia pestis, the etiological agent of plague. We used bar-tagged DNA pyrosequencing to characterize bacterial communities of wild, uninfected controls and infected fleas. Bacterial communities within Y. pestis-infected fleas were substantially more similar to one another than communities within wild or control fleas, suggesting that infection alters the bacterial community in a directed manner such that specific bacterial lineages are severely reduced in abundance or entirely eliminated from the community. Laboratory conditions also significantly altered flea-associated bacterial communities relative to wild communities, but much less so than Y. pestis infection. The abundance of Firmicutes decreased considerably in infected fleas, and Bacteroidetes were almost completely eliminated from both the control and infected fleas. Bartonella and Wolbachia were unaffected or responded positively to Y. pestis infection.
Asunto(s)
Bartonella/crecimiento & desarrollo , Siphonaptera/microbiología , Wolbachia/crecimiento & desarrollo , Yersinia pestis/fisiología , Animales , Infestaciones por Pulgas/veterinaria , SciuridaeRESUMEN
Yersinia pestis, the causative agent of plague, is primarily a rodent-associated, flea-borne zoonosis maintained in sylvatic foci throughout western North America. Transmission to humans is mediated most commonly by the flea vector Oropsylla montana and occurs predominantly in the southwestern United States. With few exceptions, previous studies showed O. montana to be an inefficient vector at transmitting Y. pestis at ambient temperatures, particularly when such fleas were fed on susceptible hosts more than a few days after ingesting an infectious blood meal. We examined whether holding fleas at subambient temperatures affected the transmissibility of Y. pestis by this vector. An infectious blood meal containing a virulent Y. pestis strain (CO96-3188) was given to colony-reared O. montana fleas. Potentially infected fleas were maintained at different temperatures (6°C, 10°C, 15°C, or 23°C). Transmission efficiencies were tested by allowing up to 15 infectious fleas to feed on each of 7 naïve CD-1 mice on days 1-4, 7, 10, 14, 17, and 21 postinfection (p.i.). Mice were monitored for signs of infection for 21 days after exposure to infectious fleas. Fleas held at 6°C, 10°C, and 15°C were able to effectively transmit at every time point p.i. The percentage of transmission to naïve mice by fleas maintained at low temperatures (46.0% at 6°C, 71.4% at 10°C, 66.7% at 15°C) was higher than for fleas maintained at 23°C (25.4%) and indicates that O. montana fleas efficiently transmit Y. pestis at low temperatures. Moreover, pooled percent per flea transmission efficiencies for flea cohorts maintained at temperatures of 10°C and 15°C (8.67% and 7.87%, respectively) showed a statistically significant difference in the pooled percent per flea transmission efficiency from fleas maintained at 23°C (1.94%). This is the first comprehensive study to demonstrate efficient transmission of Y. pestis by O. montana fleas maintained at temperatures as low as 6°C. Our findings further contribute to the understanding of plague ecology in temperate climates by providing support for the hypothesis that Y. pestis is able to overwinter within the flea gut and potentially cause infection during the following transmission season. The findings also might hold implications for explaining the focality of plague in tropical regions.
Asunto(s)
Infestaciones por Pulgas/parasitología , Insectos Vectores/microbiología , Peste/transmisión , Siphonaptera/microbiología , Yersinia pestis/fisiología , Animales , Reservorios de Enfermedades , Femenino , Humanos , Masculino , Ratones , Peste/microbiología , Estaciones del Año , Organismos Libres de Patógenos Específicos , Temperatura , Yersinia pestis/patogenicidad , ZoonosisRESUMEN
BACKGROUND: Traditionally, efficient flea-borne transmission of Yersinia pestis, the causative agent of plague, was thought to be dependent on a process referred to as blockage in which biofilm-mediated growth of the bacteria physically blocks the flea gut, leading to the regurgitation of contaminated blood into the host. This process was previously shown to be temperature-regulated, with blockage failing at temperatures approaching 30°C; however, the abilities of fleas to transmit infections at different temperatures had not been adequately assessed. We infected colony-reared fleas of Xenopsylla cheopis with a wild type strain of Y. pestis and maintained them at 10, 23, 27, or 30°C. Naïve mice were exposed to groups of infected fleas beginning on day 7 post-infection (p.i.), and every 3-4 days thereafter until day 14 p.i. for fleas held at 10°C, or 28 days p.i. for fleas held at 23-30°C. Transmission was confirmed using Y. pestis-specific antigen or antibody detection assays on mouse tissues. RESULTS: Although no statistically significant differences in per flea transmission efficiencies were detected between 23 and 30°C, efficiencies were highest for fleas maintained at 23°C and they began to decline at 27 and 30°C by day 21 p.i. These declines coincided with declining median bacterial loads in fleas at 27 and 30°C. Survival and feeding rates of fleas also varied by temperature to suggest fleas at 27 and 30°C would be less likely to sustain transmission than fleas maintained at 23°C. Fleas held at 10°C transmitted Y. pestis infections, although flea survival was significantly reduced compared to that of uninfected fleas at this temperature. Median bacterial loads were significantly higher at 10°C than at the other temperatures. CONCLUSIONS: Our results suggest that temperature does not significantly effect the per flea efficiency of Y. pestis transmission by X. cheopis, but that temperature is likely to influence the dynamics of Y. pestis flea-borne transmission, perhaps by affecting persistence of the bacteria in the flea gut or by influencing flea survival. Whether Y. pestis biofilm production is important for transmission at different temperatures remains unresolved, although our results support the hypothesis that blockage is not necessary for efficient transmission.
Asunto(s)
Insectos Vectores/fisiología , Peste/transmisión , Xenopsylla/fisiología , Yersinia pestis/fisiología , Animales , Femenino , Infestaciones por Pulgas/parasitología , Humanos , Insectos Vectores/microbiología , Masculino , Ratones , Peste/microbiología , Peste/parasitología , Xenopsylla/microbiologíaRESUMEN
In recent decades, the majority of human plague cases (caused by Yersinia pestis) have been reported from Africa. In northwest Uganda, which has had recent plague outbreaks, cat fleas (Ctenocephalides felis) have been reported as the most common fleas in the home environment, which is suspected to be a major exposure site for human plague in this country. In the past, C. felis has been viewed as only a nuisance-biting insect because limited laboratory studies suggested it is incapable of transmitting Y. pestis or is an inefficient vector. Our laboratory study shows that C. felis is a competent vector of plague bacteria, but that efficiency is low compared with another flea species collected in the same area: the oriental rat flea, Xenopsylla cheopis. On the other hand, despite its low vector efficiency, C. felis is the most common flea in human habitations in a plague-endemic region of Uganda (Arua and Nebbi Districts), and occasionally infests potential rodent reservoirs of Y. pestis such as the roof rat (Rattus rattus) or the Nile rat (Arvicanthis niloticus). Plague control programs in this region should remain focused on reducing rat flea populations, although our findings imply that cat fleas should not be ignored by these programs as they could play a significant role as secondary vectors.
Asunto(s)
Enfermedades Endémicas , Insectos Vectores , Peste/transmisión , Siphonaptera/microbiología , Yersinia pestis/patogenicidad , Animales , Gatos , Humanos , Ratones , Peste/epidemiología , Uganda/epidemiologíaRESUMEN
Bacillus anthracis, a bioterrorism threat as well as an agricultural concern, has complex mechanisms for regulation of its major virulence factors. Genome searches identified the putative two-component system that we designated Bacillus anthracis respiratory response (Brr)A-BrrB. A brrA deletion strain was constructed, and real-time reverse transcriptase polymerase chain reaction and Western blot analysis were used to assess the effect of BrrA-BrrB on levels of virulence factors, the regulator atxA, and growth characteristics. When brrA was deleted, the genes for anthrax toxins (lethal factor, protective antigen, and edema factor) where expressed 4-6-log10-fold less than in the parent Sterne strain. The global regulator atxA was downregulated when compared to atxA in the Sterne strain. Thus, the BrrA-BrrB two-component system positively regulates B. anthracis toxin genes as well as the atxA regulator. Aerobic growth was not affected by the DeltabrrA mutation, but colonies showed differences in morphology, the mutant did not sporulate, and the strain lost the ability to synthesize cytochrome aa3. Gel-shift mobility assays demonstrated that BrrA bound to the promoters of genes for both protective antigen and cytochrome aa3, demonstrating that BrrA is a transcription factor. BrrA-BrrB has sequence similarity with the virulence regulator SrrA-SrrB in Staphylococcus aureus and the aerobic/anaerobic regulator, ResD-ResE, in B. subtilis, and appears to share regulatory mechanisms with ResD-ResE.
Asunto(s)
Bacillus anthracis/genética , Bacillus anthracis/patogenicidad , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Genes Bacterianos , Factores de Virulencia , Secuencia de Aminoácidos , Antígenos Bacterianos/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/fisiología , Toxinas Bacterianas/genética , Eliminación de Gen , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Virulencia/genéticaRESUMEN
The SrrAB system regulates metabolism and virulence factors in Staphylococcus aureus. We sequenced the srrAB loci of 21 isolates and performed a phylogenetic analysis. Vaginal and bovine isolates clustered together, while skin isolates were genetically diverse. Few nucleotide polymorphisms were observed, and most were synonymous. Two strains (N2 and N19) with N-terminal truncations in SrrA displayed defects in growth and abnormally upregulated virulence factor expression under low-oxygen conditions.
Asunto(s)
Proteínas Bacterianas/genética , Proteínas Bacterianas/fisiología , Proteínas Represoras/genética , Proteínas Represoras/fisiología , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/genética , Factores de Virulencia/biosíntesis , Animales , Bovinos , Enfermedades de los Bovinos/microbiología , Femenino , Humanos , Filogenia , Polimorfismo Genético , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Piel/microbiología , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus/patogenicidad , Vagina/microbiología , VirulenciaRESUMEN
Anthrax, caused by Bacillus anthracis, has been brought to the public's attention because of the 2001 bioterrorism attacks. However, anthrax is a disease that poses agricultural threats in the United States as well as human populations in Europe, China, Africa, and Australia. Glycerol monolaurate (GML) is a compound that has been shown to inhibit exotoxin production by Staphylococcus aureus and other gram-positive bacteria. Here, we study the effects of GML on growth and toxin production in B. anthracis. The Sterne strain of B. anthracis was grown to post-exponential phase with 0-, 10-, 15-, or 20-microg/ml concentrations of GML and then assayed quantitatively for protective antigen (PA) and lethal factor (LF). After 8 h, GML at concentrations greater than 20 microg/ml was bacteriostatic to growth of the organism. However, a 10-microg/ml concentration of GML was not growth inhibitory, but amounts of PA and LF made were greatly reduced. This effect was not global for all proteins when total secreted protein from culture fluids was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Through quantitative reverse transcription-PCR assays, this toxin-inhibitory effect was shown to occur at the transcriptional level, since amounts of mRNA for pagA (PA), lef (LF), and cya (edema factor) were reduced. Surprisingly, mRNA levels of atxA, a regulator of exotoxin gene expression, rose in the presence of GML. These data will be useful in developing therapeutic tools to treat anthrax disease, whether in animals or humans. These results also suggest that mechanisms of virulence regulation exist independent of atxA.