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1.  Postmortem Diagnosis of Invasive Meningococcal Disease 
Emerging Infectious Diseases  2014;20(3):453-455.
We diagnosed invasive meningococcal disease by using immunohistochemical staining of embalmed tissue and PCR of vitreous humor from 2 men in New York City. Because vitreous humor is less subject than other body fluids to putrefaction, it is a good material for postmortem analysis.
doi:10.3201/eid2003.131245
PMCID: PMC3944856  PMID: 24565379
Neisseria meningitidis; New York City; polymerase chain reaction; PCR; bacteria; New York
2.  Poxvirus Viability and Signatures in Historical Relics 
Emerging Infectious Diseases  2014;20(2):177-184.
Although it has been >30 years since the eradication of smallpox, the unearthing of well-preserved tissue material in which the virus may reside has called into question the viability of variola virus decades or centuries after its original occurrence. Experimental data to address the long-term stability and viability of the virus are limited. There are several instances of well-preserved corpses and tissues that have been examined for poxvirus viability and viral DNA. These historical specimens cause concern for potential exposures, and each situation should be approached cautiously and independently with the available information. Nevertheless, these specimens provide information on the history of a major disease and vaccination against it.
doi:10.3201/eid2002.131098
PMCID: PMC3901489  PMID: 24447382
smallpox; smallpox virus; poxvirus; viruses; variola; orthopoxvirus; vaccinia; viability; signatures; artifacts; historical relics; mummies
3.  Quantification of the Adenylate Cyclase Toxin of Bordetella pertussis In Vitro and during Respiratory Infection 
Infection and Immunity  2013;81(5):1390-1398.
Whooping cough results from infection of the respiratory tract with Bordetella pertussis, and the secreted adenylate cyclase toxin (ACT) is essential for the bacterium to establish infection. Despite extensive study of the mechanism of ACT cytotoxicity and its effects over a range of concentrations in vitro, ACT has not been observed or quantified in vivo, and thus the concentration of ACT at the site of infection is unknown. The recently developed baboon model of infection mimics the prolonged cough and transmissibility of pertussis, and we hypothesized that measurement of ACT in nasopharyngeal washes (NPW) from baboons, combined with human and in vitro data, would provide an estimate of the ACT concentration in the airway during infection. NPW contained up to ∼108 CFU/ml B. pertussis and 1 to 5 ng/ml ACT at the peak of infection. Nasal aspirate specimens from two human infants with pertussis contained bacterial concentrations similar to those in the baboons, with 12 to 20 ng/ml ACT. When ∼108 CFU/ml of a laboratory strain of B. pertussis was cultured in vitro, ACT production was detected in 60 min and reached a plateau of ∼60 ng/ml in 6 h. Furthermore, when bacteria were brought into close proximity to target cells by centrifugation, intoxication was increased 4-fold. Collectively, these data suggest that at the bacterium-target cell interface during infection of the respiratory tract, the concentration of ACT can exceed 100 ng/ml, providing a reference point for future studies of ACT and pertussis pathogenesis.
doi:10.1128/IAI.00110-13
PMCID: PMC3648026  PMID: 23429530
4.  Cell Culture and Electron Microscopy for Identifying Viruses in Diseases of Unknown Cause 
Emerging Infectious Diseases  2013;19(6):864-869.
During outbreaks of infectious diseases or in cases of severely ill patients, it is imperative to identify the causative agent. This report describes several events in which virus isolation and identification by electron microscopy were critical to initial recognition of the etiologic agent, which was further analyzed by additional laboratory diagnostic assays. Examples include severe acute respiratory syndrome coronavirus, and Nipah, lymphocytic choriomeningitis, West Nile, Cache Valley, and Heartland viruses. These cases illustrate the importance of the techniques of cell culture and electron microscopy in pathogen identification and recognition of emerging diseases.
doi:10.3201/eid1906.130173
PMCID: PMC3713842  PMID: 23731788
Viruses; electron microscopy; cell culture; emerging diseases; SARS coronavirus; Nipah virus; lymphocytic choriomeningitis virus; West Nile virus; Cache Valley virus; Heartland virus
5.  Fatal Rocky Mountain Spotted Fever in the United States, 1999–2007 
Death from Rocky Mountain spotted fever (RMSF) is preventable with prompt, appropriate treatment. Data from two independent sources were analyzed to estimate the burden of fatal RMSF and identify risk factors for fatal RMSF in the United States during 1999–2007. Despite increased reporting of RMSF cases to the Centers for Disease Control and Prevention, no significant changes in the estimated number of annual fatal RMSF cases were found. American Indians were at higher risk of fatal RMSF relative to whites (relative risk [RR] = 3.9), and children 5–9 years of age (RR = 6.0) and adults ≥ 70 years of age (RR = 3.0) were also at increased risk relative to other ages. Persons with cases of RMSF with an immunosuppressive condition were at increased risk of death (RR = 4.4). Delaying treatment of RMSF was also associated with increased deaths. These results may indicate a gap between recommendations and practice.
doi:10.4269/ajtmh.2012.11-0453
PMCID: PMC3403778  PMID: 22492159
6.  Experimental infection of cotton rats and bobwhite quail with Rickettsia parkeri 
Parasites & Vectors  2013;6:70.
Background
Amblyomma maculatum is the primary vector for Rickettsia parkeri, a spotted fever group rickettsia (SFGR) and human pathogen. Cotton rats and quail are known hosts for larval and nymphal A. maculatum; however, the role of these hosts in the ecology of R. parkeri is unknown.
Methods
Cotton rats and quail were inoculated with low or high doses of R. parkeri (strain Portsmouth) grown in Vero cells to evaluate infection by R. parkeri in these two hosts species. Animals were euthanized 2, 4, 7, 10, and 14 days post-injection (dpi) and blood, skin, and spleen samples were collected to analyze by Vero cell culture and polymerase chain reaction (PCR). In a second trial, cotton rats and quail were inoculated with R. parkeri and nymphal A. maculatum ticks were allowed to feed on animals. Animals were euthanized on 14, 20, 28, 31, and 38 dpi and blood and tissues were collected for serology and PCR assays. Fed ticks were tested for R. parkeri by PCR and Vero cell culture.
Results
Rickettsia parkeri was isolated in cell culture and detected by PCR in skin, blood, and spleen tissues of cotton rats in the initial trial 2, 4, and 7 dpi, but not in quail tissues. In the second trial, no ticks tested positive for R. parkeri by PCR or cell culture.
Conclusions
These studies demonstrate that viable R. parkeri rickettsiae can persist in the tissues of cotton rats for at least 7 days following subcutaneous inoculation of these bacteria; however, quail are apparently resistant to infection. Rickettsia parkeri was not detected in nymphal ticks that fed on R. parkeri-inoculated cotton rats or quail, suggesting an alternate route of transmission to naïve ticks.
doi:10.1186/1756-3305-6-70
PMCID: PMC3606396  PMID: 23497681
Rickettsia parkeri; Experimental infection; Cotton rat; Quail
7.  Delivery of Bordetella pertussis adenylate cyclase toxin to target cells via outer membrane vesicles 
Febs Letters  2012;586(4):459-465.
B.pertussis adenylate cyclase toxin (ACT) intoxicates cells by producing intracellular cAMP. B.pertussis outer membrane vesicles (OMV) contain ACT on their surface (OMV-ACT), but the properties of OMV-ACT were previously unknown. We found that B.pertussis in the lung from a fatal pertussis case contains OMV, suggesting an involvement in pathogenesis. OMV-ACT and ACT intoxicate cells with and without the toxin’s receptor CD11b/CD18. Intoxication by ACT is blocked by antitoxin and anti-CD11b antibodies, but not by cytochalasin-D; in contrast, OMV-ACT is unaffected by either antibody and blocked by cytochalasin-D. Thus OMV-ACT can deliver ACT by processes distinct from those of ACT alone.
doi:10.1016/j.febslet.2012.01.032
PMCID: PMC3288663  PMID: 22289177
Bordetella; outer membrane vesicles; adenylate cyclase toxin; cAMP; intoxication; endocytosis
8.  Molecular Detection and Typing of Dengue Viruses from Archived Tissues of Fatal Cases by RT-PCR and Sequencing: Diagnostic and Epidemiologic Implications 
Diagnosis of dengue virus (DENV) infection in fatal cases is challenging because of the frequent unavailability of blood or fresh tissues. For formalin-fixed, paraffin-embedded (FFPE) tissues immunohistochemistry (IHC) can be used; however, it may not be as sensitive and serotyping is not possible. The application of reverse transcription-polymerase chain reaction (RT-PCR) for the detection of DENV in FFPE tissues has been very limited. We evaluated FFPE autopsy tissues of 122 patients with suspected DENV infection by flavivirus and DENV RT-PCR, sequencing, and DENV IHC. The DENV was detected in 61 (50%) cases by RT-PCR or IHC. The RT-PCR and sequencing detected DENV in 60 (49%) cases (DENV-1 in 16, DENV-2 in 27, DENV-3 in 8, and DENV-4 in 6 cases). No serotype could be identified in three cases. The IHC detected DENV antigens in 50 (40%) cases. The RT-PCR using FFPE tissue improves detection of DENV in fatal cases and provides sequence information useful for typing and epidemiologic studies.
doi:10.4269/ajtmh.2012.11-0346
PMCID: PMC3269289  PMID: 22302871
9.  High Rates of Rickettsia parkeri Infection in Gulf Coast Ticks (Amblyomma maculatum) and Identification of “Candidatus Rickettsia Andeanae” from Fairfax County, Virginia 
Vector Borne and Zoonotic Diseases  2011;11(12):1535-1539.
Abstract
The Gulf Coast tick, Amblyomma maculatum, is a vector of Rickettsia parkeri, a recently identified human pathogen that causes a disease with clinical symptoms that resemble a mild form of Rocky Mountain spotted fever. Because the prevalence of R. parkeri infection in geographically distinct populations of A. maculatum is not fully understood, A. maculatum specimens collected as part of a tick and pathogen surveillance system in Fairfax County, Virginia, were screened to determine pathogen infection rates. Overall, R. parkeri was found in 41.4% of the A. maculatum that were screened. Additionally, the novel spotted fever group Rickettsia sp., tentatively named “Candidatus Rickettsia andeanae,” was observed for the first time in Virginia.
doi:10.1089/vbz.2011.0654
PMCID: PMC3231788  PMID: 21867421
Amblyomma maculatum; Rickettsia andeanae; Rickettsia parkeri; Virginia
10.  Gastrointestinal basidiobolomycosis treated with posaconazole 
A 67 year-old Caucasian male from Arizona presented with indolent symptoms of intestinal obstruction and hydronephrosis, found at surgery to be caused by a mass involving the terminal ileum and cecum, extending into the posterior abdominal wall and obstructing the right ureter. Histopathology was diagnostic of basidiobolomycosis. PCR of tissue and sequencing identified the fungus as, Basidiobolus ranarum. During one year of posaconazole treatment, the residual mass shrank, hydronephrosis was relieved and peripheral eosinophilia resolved.
doi:10.1016/j.mmcr.2012.11.001
PMCID: PMC3885966  PMID: 24432205
Basidiobolomycosis; Posaconazole
11.  Susceptibility of Inbred Mice to Rickettsia parkeri 
Infection and Immunity  2012;80(5):1846-1852.
Rickettsia parkeri, a member of the spotted fever group Rickettsia, is the causative agent of American boutonneuse fever in humans. Despite the increased recognition of human cases, limited information is available regarding the infection of invertebrate and vertebrate hosts for this emerging tick-borne disease. Toward the development of a viable transmission model and to further characterize the pathology associated with R. parkeri infection, inbred mouse strains (A/J, BALB/c, C3H/HeJ, and C3H/HeN) were intravenously and intradermally inoculated with 105 low-passage-number R. parkeri (Portsmouth strain), and infection, gross pathology, and histopathology were scored. Additionally, a quantitative real-time PCR (qPCR) was performed to estimate rickettsial load in heart, lung, spleen, and liver tissues of infected mice at 19 days postinoculation. Of the A/J, BALB/c, and C3H/HeN mice, none displayed universal pathology consistent with sustained infection. Compared to age-matched control mice, the intravenously inoculated C3H/HeJ mice exhibited marked facial edema and marked splenomegaly upon gross examination, while the intradermally inoculated mice developed characteristic eschar-like lesions. The C3H/HeJ mice also exhibited the greatest concentrations of rickettsial DNA from heart, lung, liver, and spleen samples when examined by qPCR. The similarity of the pathology of human disease and sustained infection suggests that the C3H/HeJ strain of mice is a promising candidate for subsequent experiments to examine the tick transmission, dissemination, and pathology of R. parkeri rickettsiosis.
doi:10.1128/IAI.00109-12
PMCID: PMC3347444  PMID: 22392926
12.  Rickettsia parkeri and Candidatus Rickettsia andeanae in Gulf Coast Ticks, Mississippi, USA 
Emerging Infectious Diseases  2012;18(10):1705-1707.
doi:10.3201/eid1810.120250
PMCID: PMC3471625  PMID: 23018026
Amblyomma maculatum; co-infection; Mississippi; spotted fever group rickettsiae; Rickettsia parkeri; Candidatus Rickettsia andeanae; bacteria; Gulf Coast ticks; Rickettsia; United States; ticks; vector-borne infections
13.  Rickettsia parkeri in Argentina 
Emerging Infectious Diseases  2008;14(12):1894-1897.
Clinical reports of an eschar-associated rickettsiosis in the Paraná River Delta of Argentina prompted an evaluation of Amblyomma triste ticks in this region. When evaluated by PCR, 17 (7.6%) of 223 questing adult A. triste ticks, collected from 2 sites in the lower Paraná River Delta, contained DNA of Rickettsia parkeri.
doi:10.3201/eid1412.080860
PMCID: PMC2634642  PMID: 19046514
Rickettsia parkeri; Amblyomma triste; rickettsiosis; Argentina; dispatch
14.  Solid Organ Transplant–associated Lymphocytic Choriomeningitis, United States, 2011 
Emerging Infectious Diseases  2012;18(8):1256-1262.
Lymphocytic choriomeningitis virus (LCMV) is carried by rodents. In very rare instances, it has been transmitted from person-to-person by organ transplantation. In 2011, a total of 14 organ recipients were infected with the virus, of which 11 died in the United States. The 4 most recent patients received organs from the same donor, which resulted in 2 deaths. Only after these 4 organ recipients became sick was it discovered that the donor had been exposed to rodents. Had this exposure been known before transplantation, the organ recipients may have been more closely monitored. Early diagnosis and treatment might have improved their chances of survival. Although organ donor screening reduces the risk for transmission of some viruses, it is not possible to screen for all possible viruses, including LCMV. For patients who get severely ill after receiving a transplant, clinicians should add LCMV infection to their list of possible causes.
Three clusters of organ transplant–associated lymphocytic choriomeningitis virus (LCMV) transmissions have been identified in the United States; 9 of 10 recipients died. In February 2011, we identified a fourth cluster of organ transplant–associated LCMV infections. Diabetic ketoacidosis developed in the organ donor in December 2010; she died with generalized brain edema after a short hospitalization. Both kidneys, liver, and lung were transplanted to 4 recipients; in all 4, severe posttransplant illness developed; 2 recipients died. Through multiple diagnostic methods, we identified LCMV infection in all persons, including in at least 1 sample from the donor and 4 recipients by reverse transcription PCR, and sequences of a 396-bp fragment of the large segment of the virus from all 5 persons were identical. In this cluster, all recipients developed severe illness, but 2 survived. LCMV infection should be considered as a possible cause of severe posttransplant illness.
doi:10.3201/eid1808.120212
PMCID: PMC3414043  PMID: 22839997
LCMV; lymphocytic choriomeningitis; organ transplant; transplant-associated infection; organ donation; viruses; zoonoses
15.  Gulf Coast Ticks (Amblyomma maculatum) and Rickettsia parkeri, United States 
Emerging Infectious Diseases  2007;13(5):751-753.
Geographic distribution of Rickettsia parkeri in its US tick vector, Amblyomma maculatum, was evaluated by PCR. R. parkeri was detected in ticks from Florida, Georgia, Kentucky, Mississippi, Oklahoma, and South Carolina, which suggests that A. maculatum may be responsible for additional cases of R. parkeri rickettsiosis throughout much of its US range.
doi:10.3201/eid1305.061468
PMCID: PMC2738460  PMID: 17553257
Rickettsia parkeri; Amblyomma maculatum; spotted fever rickettsia; dispatch
16.  Rapid, Simultaneous Detection of Clostridium sordellii and Clostridium perfringens in Archived Tissues by a Novel PCR-Based Microsphere Assay: Diagnostic Implications for Pregnancy-Associated Toxic Shock Syndrome Cases 
Clostridium sordellii and Clostridium perfringens are infrequent human pathogens; however, the case-fatality rates for the infections are very high, particularly in obstetric C. sordellii infections (>90%). Deaths from Clostridium sordellii and Clostridium perfringens toxic shock (CTS) are sudden, and diagnosis is often challenging. Formalin-fixed, paraffin-embedded (FFPE) tissues usually are the only specimens available for sudden fatal cases, and immunohistochemistry (IHC) for Clostridia is generally performed but it cannot identify species. A clear need exists for a rapid, species-specific diagnostic assay for FFPE tissues. We developed a duplex PCR-based microsphere assay for simultaneous detection of C. sordellii and C. perfringens and evaluated DNA extracted from 42 Clostridium isolates and FFPE tissues of 28 patients with toxic shock/endometritis (20 CTS, 8 non-CTS, as confirmed by PCR and sequencing). The microsphere assay correctly identified C. sordellii and C. perfringens in all known isolates and in all CTS patients (10 C. sordellii, 8 C. perfringens, 2 both) and showed 100% concordance with PCR and sequencing results. The microsphere assay is a rapid, specific, and cost-effective method for the diagnosis of CTS and offers the advantage of simultaneous testing for C. sordellii and C. perfringens in FFPE tissues using a limited amount of DNA.
doi:10.1155/2012/972845
PMCID: PMC3320015  PMID: 22536012
17.  Clinical, Epidemiologic, Histopathologic and Molecular Features of an Unexplained Dermopathy 
PLoS ONE  2012;7(1):e29908.
Background
Morgellons is a poorly characterized constellation of symptoms, with the primary manifestations involving the skin. We conducted an investigation of this unexplained dermopathy to characterize the clinical and epidemiologic features and explore potential etiologies.
Methods
A descriptive study was conducted among persons at least 13 years of age and enrolled in Kaiser Permanente Northern California (KPNC) during 2006–2008. A case was defined as the self-reported emergence of fibers or materials from the skin accompanied by skin lesions and/or disturbing skin sensations. We collected detailed epidemiologic data, performed clinical evaluations and geospatial analyses and analyzed materials collected from participants' skin.
Results
We identified 115 case-patients. The prevalence was 3.65 (95% CI = 2.98, 4.40) cases per 100,000 enrollees. There was no clustering of cases within the 13-county KPNC catchment area (p = .113). Case-patients had a median age of 52 years (range: 17–93) and were primarily female (77%) and Caucasian (77%). Multi-system complaints were common; 70% reported chronic fatigue and 54% rated their overall health as fair or poor with mean Physical Component Scores and Mental Component Scores of 36.63 (SD = 12.9) and 35.45 (SD = 12.89), respectively. Cognitive deficits were detected in 59% of case-patients and 63% had evidence of clinically significant somatic complaints; 50% had drugs detected in hair samples and 78% reported exposure to solvents. Solar elastosis was the most common histopathologic abnormality (51% of biopsies); skin lesions were most consistent with arthropod bites or chronic excoriations. No parasites or mycobacteria were detected. Most materials collected from participants' skin were composed of cellulose, likely of cotton origin.
Conclusions
This unexplained dermopathy was rare among this population of Northern California residents, but associated with significantly reduced health-related quality of life. No common underlying medical condition or infectious source was identified, similar to more commonly recognized conditions such as delusional infestation.
doi:10.1371/journal.pone.0029908
PMCID: PMC3266263  PMID: 22295070
18.  Rickettsia parkeri in Amblyomma maculatum Ticks, North Carolina, USA, 2009–2010 
Emerging Infectious Diseases  2011;17(12):2350-2353.
We detected Rickettsia parkeri in 20%−33% of Amblyomma maculatum ticks sampled in North Carolina. Results highlight the high frequencies of R. parkeri–infected ticks in the state with the highest annual incidence of Rocky Mountain spotted fever. Epidemiologic studies are needed to definitively link R. parkeri to cases of spotted fever rickettsiosis.
doi:10.3201/eid1712.110789
PMCID: PMC3311201  PMID: 22172164
Spotted fever group rickettsiae; Rickettsia; vector-borne infections; Rocky Mountain spotted fever; North Carolina; ticks; Candidatus Rickettsia andeanae; Rickettsia parkeri; Amblyomma maculatum
19.  Ultrastructural Characterization of Pandemic (H1N1) 2009 Virus 
Emerging Infectious Diseases  2011;17(11):2056-2059.
We evaluated pandemic influenza A (H1N1) 2009 virus isolates and respiratory tissues collected at autopsy by electron microscopy. Many morphologic characteristics were similar to those previously described for influenza virus. One of the distinctive features was dense tubular structures in the nuclei of infected cells.
doi:10.3201/eid1711.110258
PMCID: PMC3310559  PMID: 22099097
viruses; pandemic; pandemic (H1N1) 2009; influenza; electron microscopy; immunoelectron microscopy; viral proteins; dispatch
20.  Rickettsia parkeri Rickettsiosis, Argentina 
Emerging Infectious Diseases  2011;17(7):1169-1173.
Rickettsia parkeri, a recently identified cause of spotted fever rickettsiosis in the United States, has been found in Amblyomma triste ticks in several countries of South America, including Argentina, where it is believed to cause disease in humans. We describe the clinical and epidemiologic characteristics of 2 patients in Argentina with confirmed R. parkeri infection and 7 additional patients with suspected R. parkeri rickettsiosis identified at 1 hospital during 2004–2009. The frequency and character of clinical signs and symptoms among these 9 patients closely resembled those described for patients in the United States (presence of an inoculation eschar, maculopapular rash often associated with pustules or vesicles, infrequent gastrointestinal manifestations, and relatively benign clinical course). Many R. parkeri infections in South America are likely to be misdiagnosed as other infectious diseases, including Rocky Mountain spotted fever, dengue, or leptospirosis.
doi:10.3201/eid1707.101857
PMCID: PMC3381406  PMID: 21762568
21.  Pandemic (H1N1) 2009 Virus in 3 Wildlife Species, San Diego, California, USA 
Emerging Infectious Diseases  2011;17(4):747-749.
doi:10.3201/eid1704.101355
PMCID: PMC3377413  PMID: 21470480
Viruses; influenza; pandemic (H1N1) 2009; respiratory infections; California; zoological gardens; wildlife; reservoirs; H1N1; letter
22.  Eschar-associated Spotted Fever Rickettsiosis, Bahia, Brazil 
Emerging Infectious Diseases  2011;17(2):275-278.
In Brazil, Brazilian spotted fever was once considered the only tick-borne rickettsial disease. We report eschar-associated rickettsial disease that occurred after a tick bite. The etiologic agent is most related to Rickettsia parkeri, R. africae, and R. sibirica and probably widely distributed from São Paulo to Bahia in the Atlantic Forest.
doi:10.3201/eid1702.100859
PMCID: PMC3204763  PMID: 21291605
Spotted fever group rickettsiosis; rickettsia; eschar; multiple-locus sequence analysis; ticks; molecular diagnosis; dispatch
23.  Blastomycosis in Man after Kinkajou Bite 
Emerging Infectious Diseases  2011;17(2):268-270.
We report transmission of Blastomyces dermatitidis fungal infection from a pet kinkajou to a man. When treating a patient with a recalcitrant infection and a history of an animal bite, early and complete animal necropsy and consideration of nonbacterial etiologies are needed.
doi:10.3201/eid1702.101046
PMCID: PMC3204770  PMID: 21291603
Blastomycosis; zoonoses; wound infection; fungi; kinkajou; human infection; dispatch
24.  Isolation of Rickettsia parkeri and Identification of a Novel Spotted Fever Group Rickettsia sp. from Gulf Coast Ticks (Amblyomma maculatum) in the United States▿  
Until recently, Amblyomma maculatum (the Gulf Coast tick) had garnered little attention compared to other species of human-biting ticks in the United States. A. maculatum is now recognized as the principal vector of Rickettsia parkeri, a pathogenic spotted fever group rickettsia (SFGR) that causes an eschar-associated illness in humans that resembles Rocky Mountain spotted fever. A novel SFGR, distinct from other recognized Rickettsia spp., has also been detected recently in A. maculatum specimens collected in several regions of the southeastern United States. In this study, 198 questing adult Gulf Coast ticks were collected at 4 locations in Florida and Mississippi; 28% of these ticks were infected with R. parkeri, and 2% of these were infected with a novel SFGR. Seventeen isolates of R. parkeri from individual specimens of A. maculatum were cultivated in Vero E6 cells; however, all attempts to isolate the novel SFGR were unsuccessful. Partial genetic characterization of the novel SFGR revealed identity with several recently described, incompletely characterized, and noncultivated SFGR, including “Candidatus Rickettsia andeanae” and Rickettsia sp. Argentina detected in several species of Neotropical ticks from Argentina and Peru. These findings suggest that each of these “novel” rickettsiae represent the same species. This study considerably expanded the number of low-passage, A. maculatum-derived isolates of R. parkeri and characterized a second, sympatric Rickettsia sp. found in Gulf Coast ticks.
doi:10.1128/AEM.02737-09
PMCID: PMC2863434  PMID: 20208020
25.  Wide Dispersal and Possible Multiple Origins of Low-Copy-Number Plasmids in Rickettsia Species Associated with Blood-Feeding Arthropods▿  
Plasmids are mobile genetic elements of bacteria that can impart important adaptive traits, such as increased virulence or antibiotic resistance. We report the existence of plasmids in Rickettsia (Rickettsiales; Rickettsiaceae) species, including Rickettsia akari, “Candidatus Rickettsia amblyommii,” R. bellii, R. rhipicephali, and REIS, the rickettsial endosymbiont of Ixodes scapularis. All of the rickettsiae were isolated from humans or North and South American ticks. R. parkeri isolates from both continents did not possess plasmids. We have now demonstrated plasmids in nearly all Rickettsia species that we have surveyed from three continents, which represent three of the four major proposed phylogenetic groups associated with blood-feeding arthropods. Gel-based evidence consistent with the existence of multiple plasmids in some species was confirmed by cloning plasmids with very different sequences from each of two “Ca. Rickettsia amblyommii” isolates. Phylogenetic analysis of rickettsial ParA plasmid partitioning proteins indicated multiple parA gene origins and plasmid incompatibility groups, consistent with possible multiple plasmid origins. Phylogenetic analysis of potentially host-adaptive rickettsial small heat shock proteins showed that hsp2 genes were plasmid specific and that hsp1 genes, found only on plasmids of “Ca. Rickettsia amblyommii,” R. felis, R. monacensis, and R. peacockii, were probably acquired independently of the hsp2 genes. Plasmid copy numbers in seven Rickettsia species ranged from 2.4 to 9.2 per chromosomal equivalent, as determined by real-time quantitative PCR. Plasmids may be of significance in rickettsial evolution and epidemiology by conferring genetic plasticity and host-adaptive traits via horizontal gene transfer that counteracts the reductive genome evolution typical of obligate intracellular bacteria.
doi:10.1128/AEM.02988-09
PMCID: PMC2838022  PMID: 20097813

Results 1-25 (56)