Publications by authors named "Ricardo G Maggi"

95 Publications

Comparison of Serological and Molecular Assays for Species in Dogs with Hemangiosarcoma.

Pathogens 2021 Jun 23;10(7). Epub 2021 Jun 23.

Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA.

Currently, a gold standard diagnostic test for infection in dogs is lacking. This represents a critical limitation for the development and evaluation of new diagnostic tests, as well as for the diagnosis of, and research on, bartonellosis in dogs. This retrospective observational study aims to compare the results of commonly performed and newly-reported spp. diagnostic tests in banked clinical specimens from 90 dogs with hemangiosarcoma (HSA) using composite reference standard (CRS) and random effects latent class analysis (RE-LCA) techniques. Samples from each dog were tested using six serological or molecular diagnostic assays, including indirect fluorescent antibody (IFA) and Western blot (WB) for the detection of antibodies in serum, and qPCR and droplet digital PCR (ddPCR) in blood and fresh frozen tissue biopsy samples (mainly splenic HSA tumors and histopathologically normal spleen or skin/adipose tissue). infection prevalence was estimated to be 78% based on the CRS (parallel testing with all six assays), and 64% based on the RE-LCA model. The assay with the highest diagnostic accuracy was qPCR performed on fresh frozen tissue biopsy samples (sensitivity: 94% by RE-LCA and 80% by CRS; specificity: 100%). When comparing newly-reported to traditional diagnostic assays, ddPCR was more sensitive for the detection of DNA than qPCR when testing blood samples (36% vs. 0%, < 0.0001). Dogs that were positive on serological assays alone with negative molecular assays were highly unlikely (<3%) to be classified as infected by the RE-LCA model. These data indicate that spp. DNA can be PCR amplified from fresh frozen tissues from a majority of dogs with HSA using both qPCR and ddPCR, supporting the use of these methods for future controlled studies comparing the prevalence of spp. DNA in the tissue of dogs with HSA to that of unaffected controls.
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http://dx.doi.org/10.3390/pathogens10070794DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8308881PMC
June 2021

Detected in Malignant Melanoma, a Preliminary Study.

Pathogens 2021 Mar 10;10(3). Epub 2021 Mar 10.

Department of Biomedical Sciences, Duluth Campus, Medical School, University of Minnesota, Duluth, MN 55812, USA.

, , and are bacteria known to cause verruga peruana or bacillary angiomatosis, vascular endothelial growth factor (VEGF)-dependent cutaneous lesions in humans. Given the bacteria's association with the dermal niche and clinical suspicion of occult infection by a dermatologist, we determined if patients with melanoma had evidence of spp. infection. Within a one-month period, eight patients previously diagnosed with melanoma volunteered to be tested for evidence of spp. exposure/infection. Subsequently, confocal immunohistochemistry and PCR for spp. were used to study melanoma tissues from two patients. Blood from seven of the eight patients was either seroreactive, PCR positive, or positive by both modalities for spp. exposure. Subsequently, organisms that co-localized with VEGFC immunoreactivity were visualized using multi-immunostaining confocal microscopy of thick skin sections from two patients. Using a co-culture model, was observed to enter melanoma cell cytoplasm and resulted in increased vascular endothelial growth factor C (VEGFC) and interleukin 8 (IL-8) production. Findings from this small number of patients support the need for future investigations to determine the extent to which spp. are a component of the melanoma pathobiome.
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http://dx.doi.org/10.3390/pathogens10030326DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7998106PMC
March 2021

Associated Cutaneous Lesions (BACL) in People with Neuropsychiatric Symptoms.

Pathogens 2020 Dec 4;9(12). Epub 2020 Dec 4.

Dermatology Clinic, Truman Medical Center, University of Missouri Kansas City, Kansas City, MO 64108, USA.

species are globally important emerging pathogens that were not known to infect animals or humans in North America prior to the human immunodeficiency virus (HIV) epidemic. Ongoing improvements in diagnostic testing modalities have allowed for the discovery of species (spp.) DNA in blood; cerebrospinal fluid; and the skin of patients with cutaneous lesions, fatigue, myalgia, and neurological symptoms. We describe spp. test results for participants reporting neuropsychiatric symptoms, the majority of whom reported the concurrent development of cutaneous lesions. Study participants completed a medical history, a risk factor questionnaire, and provided cutaneous lesion photographs. spp. serology and alpha proteobacteria enrichment blood culture/PCR were assessed. Within a 14-month period, 33 participants enrolled; 29/33 had serological and/or PCR evidence supporting spp. infection, of whom 24 reported concurrent cutaneous lesions since neuropsychiatric symptom onset. We conclude that cutaneous lesions were common among people reporting neuropsychiatric symptoms and spp. infection or exposure. Additional studies, using sensitive microbiological and imaging techniques, are needed to determine if, or to what extent, spp. might contribute to cutaneous lesions and neuropsychiatric symptoms in patients.
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http://dx.doi.org/10.3390/pathogens9121023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761945PMC
December 2020

Parasites and vector-borne diseases disseminated by rehomed dogs.

Parasit Vectors 2020 Nov 10;13(1):546. Epub 2020 Nov 10.

Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, Leipzig, Germany.

The Companion Vector-Borne Diseases (CVBD) World Forum is a working group of leading international experts who meet annually to evaluate current scientific findings and future trends concerning the distribution, pathogenesis, clinical presentation, diagnosis and prevention of vector-borne infections of dogs and cats. At the 14th Symposium of the CVBD World Forum in Trieste, Italy (March 25-28, 2019), we identified the need to (i) bring attention to the potential spread of parasites and vectors with relocated dogs, and (ii) provide advice to the veterinary profession regarding the importance of surveillance and treatment for parasites and vector-borne infections when rehoming dogs. This letter shares a consensus statement from the CVBD World Forum as well as a summary of the problem faced, including the role of veterinary professionals in parasite surveillance, causal issues, and the importance of interdisciplinary cooperation in addressing the problem. To limit opportunities for dissemination of parasites and vectors, whenever possible, underlying problems creating the need for dog rehoming should be addressed. However, when it is necessary to rehome dogs, this should ideally take place in the country and national region of origin. When geographically distant relocation occurs, veterinary professionals have a vital role to play in public education, vigilance for detection of exotic vectors and infections, and alerting the medical community to the risk(s) for pathogen spread. With appropriate veterinary intervention, dog welfare needs can be met without inadvertently allowing global spread of parasites and their vectors.
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http://dx.doi.org/10.1186/s13071-020-04407-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7653694PMC
November 2020

Hopping species and borders: detection of Bartonella spp. in avian nest fleas and arctic foxes from Nunavut, Canada.

Parasit Vectors 2020 Sep 14;13(1):469. Epub 2020 Sep 14.

Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, S7N 5B4, Canada.

Background: In a warmer and more globally connected Arctic, vector-borne pathogens of zoonotic importance may be increasing in prevalence in native wildlife. Recently, Bartonella henselae, the causative agent of cat scratch fever, was detected in blood collected from arctic foxes (Vulpes lagopus) that were captured and released in the large goose colony at Karrak Lake, Nunavut, Canada. This bacterium is generally associated with cats and cat fleas, which are absent from Arctic ecosystems. Arctic foxes in this region feed extensively on migratory geese, their eggs, and their goslings. Thus, we hypothesized that a nest flea, Ceratophyllus vagabundus vagabundus (Boheman, 1865), may serve as a vector for transmission of Bartonella spp.

Methods: We determined the prevalence of Bartonella spp. in (i) nest fleas collected from 5 arctic fox dens and (ii) 37 surrounding goose nests, (iii) fleas collected from 20 geese harvested during arrival at the nesting grounds and (iv) blood clots from 57 adult live-captured arctic foxes. A subsample of fleas were identified morphologically as C. v. vagabundus. Remaining fleas were pooled for each nest, den, or host. DNA was extracted from flea pools and blood clots and analyzed with conventional and real-time polymerase chain reactions targeting the 16S-23S rRNA intergenic transcribed spacer region.

Results: Bartonella henselae was identified in 43% of pooled flea samples from nests and 40% of pooled flea samples from fox dens. Bartonella vinsonii berkhoffii was identified in 30% of pooled flea samples collected from 20 geese. Both B. vinsonii berkhoffii (n = 2) and B. rochalimae (n = 1) were identified in the blood of foxes.

Conclusions: We confirm that B. henselae, B. vinsonii berkhoffii and B. rochalimae circulate in the Karrak Lake ecosystem and that nest fleas contain B. vinsonii and B. henselae DNA, suggesting that this flea may serve as a potential vector for transmission among Arctic wildlife.
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http://dx.doi.org/10.1186/s13071-020-04344-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7490881PMC
September 2020

Development and validation of a droplet digital PCR assay for the detection and quantification of Bartonella species within human clinical samples.

J Microbiol Methods 2020 09 11;176:106022. Epub 2020 Aug 11.

Galaxy Diagnostics, Inc, 7020 Kit Creek Rd #130, Research Triangle Park, NC 27709, USA. Electronic address:

This report describes the development, optimization, and validation of a ddPCR assay for the detection of Bartonella spp. DNA within several sample matrices, including clinical blood samples from patients with or without documented Bartonella spp. bacteremia. The Bartonella spp. ddPCR assay was developed based upon previously published TaqMan-based qPCR assays that can amplify DNA of over 25 Bartonella spp. Host DNA (housekeeping gene) amplification serves as a reference target to facilitate quantification. The efficiency, sensitivity, and specificity of the Bartonella spp. ddPCR assay was assessed by direct comparison with the current qPCR methods used by the Intracellular Pathogens Research Laboratory (North Carolina State University, North Carolina, USA), and Galaxy Diagnostics (Research Triangle Park, North Carolina, USA). Bartonella spp. ddPCR assay parameters were successfully optimized to detect Bartonella concentrations equivalent to 0.5 bacterial genome copies per microliter of blood (0.001 pg/ul of bacterial DNA). The number of droplets detected (resolution) for each concentration was consistent across each of four assessed time points. The Bartonella spp. ddPCR assay detected 16 species/strains including B. henselae; B. quintana; B. vinsonii subsp. berkhoffii (genotypes I, II, III and IV); B. vinsonii subsp. vinsonii; B. melophagi; B. volans; B. monaki; B. alsatica; B. bovis; B. elizabethae; B. clarridgeiae; and B. koehlerae. Bartonella DNA was detected in only one previously negative patient sample (119/120 negative; 99% specificity). The ddPCR sensitivity (53/112) was significantly better than qPCR (6/112) when testing patient blood and enrichment blood culture samples. The development of commercial ddPCR systems with integrated technologies has significantly streamlined the DNA detection process, making it more efficient and standardized for clinical diagnostic testing. The assay described in this work is the first step toward the development of a multiplex ddPCR assay (i.e., using the QX One from Bio-Rad) for the simultaneous detection and absolute quantification of multiple vector-borne pathogens (such as Babesia, Bartonella and Borrelia) within clinical samples.
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http://dx.doi.org/10.1016/j.mimet.2020.106022DOI Listing
September 2020

Molecular prevalence of Bartonella, Babesia, and hemotropic Mycoplasma species in dogs with hemangiosarcoma from across the United States.

PLoS One 2020 10;15(1):e0227234. Epub 2020 Jan 10.

Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, United States of America.

Hemangiosarcoma (HSA), a locally invasive and highly metastatic endothelial cell neoplasm, accounts for two-thirds of all cardiac and splenic neoplasms in dogs. Bartonella spp. infection has been reported in association with neoplastic and non-neoplastic vasoproliferative lesions in animals and humans. The objective of this study was to determine the prevalence of Bartonella spp. in conjunction with two other hemotropic pathogens, Babesia spp. and hemotropic Mycoplasma spp., in tissues and blood samples from 110 dogs with histopathologically diagnosed HSA from throughout the United States. This was a retrospective, observational study using clinical specimens from 110 dogs with HSA banked by the biospecimen repository of the Canine Comparative Oncology and Genomics Consortium. Samples provided for this study from each dog included: fresh frozen HSA tumor tissue (available from n = 100 of the 110 dogs), fresh frozen non-tumor tissue (n = 104), and whole blood and serum samples (n = 108 and 107 respectively). Blood and tissues were tested by qPCR for Bartonella, hemotropic Mycoplasma, and Babesia spp. DNA; serum was tested for Bartonella spp. antibodies. Bartonella spp. DNA was amplified and sequenced from 73% of dogs with HSA (80/110). In contrast, hemotropic Mycoplasma spp. DNA was amplified from a significantly smaller proportion (5%, p<0.0001) and Babesia spp. DNA was not amplified from any dog. Of the 100 HSA tumor samples submitted, 34% were Bartonella PCR positive (32% of splenic tumors, 57% of cardiac tumors, and 17% of other tumor locations). Of 104 non-tumor tissues, 63% were Bartonella PCR positive (56% of spleen samples, 93% of cardiac samples, and 63% of skin/subcutaneous samples). Of dogs with Bartonella positive HSA tumor, 76% were also positive in non-tumor tissue. Bartonella spp. DNA was not PCR amplified from whole blood. This study documented a high prevalence of Bartonella spp. DNA in dogs with HSA from geographically diverse regions of the United States. While 73% of all tissue samples from these dogs were PCR positive for Bartonella DNA, none of the blood samples were, indicating that whole blood samples do not reflect tissue presence of this pathogen. Future studies are needed to further investigate the role of Bartonella spp. in the development of HSA.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0227234PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6953799PMC
May 2020

Detection of Bartonella spp. in dogs after infection with Rickettsia rickettsii.

J Vet Intern Med 2020 Jan 31;34(1):145-159. Epub 2019 Dec 31.

Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina.

Background: Dynamics of infection by Bartonella and Rickettsia species, which are epidemiologically associated in dogs, have not been explored in a controlled setting.

Objectives: Describe an outbreak investigation of occult Bartonella spp. infection among a group of dogs, discovered after experimentally induced Rickettsia rickettsii (Rr) infection.

Animals: Six apparently healthy purpose-bred Beagles obtained from a commercial vendor.

Methods: Retrospective and prospective study. Dogs were serially tested for Bartonella spp. and Rr using serology, culture, and PCR, over 3 study phases: 3 months before inoculation with Rr (retrospective), 6 weeks after inoculation with Rr (retrospective), and 8 months of follow-up (prospective).

Results: Before Rr infection, 1 dog was Bartonella henselae (Bh) immunofluorescent antibody assay (IFA) seroreactive and 1 was Rickettsia spp. IFA seroreactive. After inoculation with Rr, all dogs developed mild Rocky Mountain spotted fever compatible with low-dose Rr infection, seroconverted to Rickettsia spp. within 4-11 days, and recovered within 1 week. When 1 dog developed ear tip vasculitis with intra-lesional Bh, an investigation of Bartonella spp. infection was undertaken. All dogs had seroconverted to 1-3 Bartonella spp. between 7 and 18 days after Rr inoculation. Between 4 and 8 months after Rr inoculation, Bh DNA was amplified from multiple tissues from 2 dogs, and Bartonella vinsonii subsp. berkhoffii (Bvb) DNA was amplified from 4 of 5 dogs' oral swabs.

Conclusions And Clinical Importance: Vector-borne disease exposure was demonstrated in research dogs from a commercial vendor. Despite limitations, our results support the possibilities of recrudescence of chronic subclinical Bartonella spp. infection after Rr infection and horizontal direct-contact transmission between dogs.
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http://dx.doi.org/10.1111/jvim.15675DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6979086PMC
January 2020

My Mother's Story: Tick Borne Ehrlichiosis and a Life Well-Lived.

Vector Borne Zoonotic Dis 2020 05 16;20(5):319-324. Epub 2019 Dec 16.

Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.

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http://dx.doi.org/10.1089/vbz.2019.2570DOI Listing
May 2020

A review on the occurrence of companion vector-borne diseases in pet animals in Latin America.

Parasit Vectors 2019 Mar 28;12(1):145. Epub 2019 Mar 28.

Institute of Parasitology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.

Companion vector-borne diseases (CVBDs) are an important threat for pet life, but may also have an impact on human health, due to their often zoonotic character. The importance and awareness of CVBDs continuously increased during the last years. However, information on their occurrence is often limited in several parts of the world, which are often especially affected. Latin America (LATAM), a region with large biodiversity, is one of these regions, where information on CVBDs for pet owners, veterinarians, medical doctors and health workers is often obsolete, limited or non-existent. In the present review, a comprehensive literature search for CVBDs in companion animals (dogs and cats) was performed for several countries in Central America (Belize, Caribbean Islands, Costa Rica, Cuba, Dominican Republic, El Salvador, Guatemala, Honduras, Mexico, Nicaragua, Panama, Puerto Rico) as well as in South America (Argentina, Bolivia, Brazil, Chile, Colombia, Ecuador, French Guiana, Guyana (British Guyana), Paraguay, Peru, Suriname, Uruguay, Venezuela) regarding the occurrence of the following parasitic and bacterial diseases: babesiosis, heartworm disease, subcutaneous dirofilariosis, hepatozoonosis, leishmaniosis, trypanosomosis, anaplasmosis, bartonellosis, borreliosis, ehrlichiosis, mycoplasmosis and rickettsiosis. An overview on the specific diseases, followed by a short summary on their occurrence per country is given. Additionally, a tabular listing on positive or non-reported occurrence is presented. None of the countries is completely free from CVBDs. The data presented in the review confirm a wide distribution of the CVBDs in focus in LATAM. This wide occurrence and the fact that most of the CVBDs can have a quite severe clinical outcome and their diagnostic as well as therapeutic options in the region are often difficult to access and to afford, demands a strong call for the prevention of pathogen transmission by the use of ectoparasiticidal and anti-feeding products as well as by performing behavioural changes.
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http://dx.doi.org/10.1186/s13071-019-3407-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6438007PMC
March 2019

Bloodstream Infection in a Boy With Pediatric Acute-Onset Neuropsychiatric Syndrome.

J Cent Nerv Syst Dis 2019 18;11:1179573519832014. Epub 2019 Mar 18.

Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA.

Background: With the advent of more sensitive culture and molecular diagnostic testing modalities, spp. infections have been documented in blood and/or cerebrospinal fluid specimens from patients with diverse neurological symptoms. Pediatric acute-onset neuropsychiatric syndrome (PANS) is characterized by an unusually abrupt onset of cognitive, behavioral, or neurological symptoms. Between October 2015 and January 2017, a 14-year-old boy underwent evaluation by multiple specialists for sudden-onset psychotic behavior (hallucinations, delusions, suicidal and homicidal ideation).

Methods: In March 2017, spp. serology (indirect fluorescent antibody assays) and polymerase chain reaction (PCR) amplification, DNA sequencing, and enrichment blood culture were used on a research basis to assess spp. exposure and bloodstream infection, respectively. PCR assays targeting other vector-borne infections were performed to assess potential co-infections.

Results: For 18 months, the boy remained psychotic despite 4 hospitalizations, therapeutic trials involving multiple psychiatric medication combinations, and immunosuppressive treatment for autoimmune encephalitis. Neurobartonellosis was diagnosed after cutaneous lesions developed. Subsequently, despite nearly 2 consecutive months of doxycycline administration, DNA was PCR amplified and sequenced from the patient's blood, and from alphaproteobacteria growth medium enrichment blood cultures. serology was negative. During treatment with combination antimicrobial chemotherapy, he experienced a gradual progressive decrease in neuropsychiatric symptoms, cessation of psychiatric drugs, resolution of -associated cutaneous lesions, and a return to all pre-illness activities.

Conclusions: This case report suggests that bloodstream infection may contribute to progressive, recalcitrant neuropsychiatric symptoms consistent with PANS in a subset of patients.
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http://dx.doi.org/10.1177/1179573519832014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6423671PMC
March 2019

Detection and Prevalence of spp. in American Black Bears () from Eastern and Western North Carolina, USA.

J Wildl Dis 2019 07 29;55(3):678-681. Epub 2019 Jan 29.

1 College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, North Carolina 27607, USA.

Blood samples collected from American black bears () in eastern and western North Carolina, US, were analyzed for piroplasms. Piroplasmids were detected in 17% (23/132) of the animals surveyed. We detected a spp. previously identified in North American raccoons () and a maned wolf (); prevalence was 22% (14/64) and 13% (9/68) in the mountain and coastal black bear populations, respectively. The presence of the same species in black bears, raccoons, and a maned wolf suggests piroplasms may not be host specific.
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http://dx.doi.org/10.7589/2018-06-164DOI Listing
July 2019

Bartonella spp. Bloodstream Infection in a Canadian Family.

Vector Borne Zoonotic Dis 2019 04 27;19(4):234-241. Epub 2018 Dec 27.

1 Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina.

Historically, Bartonella spp. have been associated with febrile illness (Oroya fever, trench fever, and cat scratch disease), endocarditis (numerous Bartonella spp.), and vasoproliferative lesions (Bartonella bacilliformis, Bartonella quintana, Bartonella henselae, and Bartonella vinsonii subsp. berkhoffii), occurring most often but not exclusively in immunocompromised patients. Recently, bloodstream infections with various Bartonella spp. have been documented in nonimmunocompromised individuals in association with a spectrum of cardiovascular, neurologic, and rheumatologic symptoms. As documented in this family, symptoms for which the medical implications remain unclear can occur in multiple family members infected with one or more Bartonella spp. Serial serologic and molecular microbiological findings supported exposure to or infection with Bartonella spp. in all seven family members. Either antibiotics failed to eliminate bacteremic infection, resulted in partial resolution of symptoms, or potentially reinfection occurred during the 19-month study period. There is a substantial need for clinical research to clarify the extent to which Bartonella spp. bacteremia induces nonspecific cardiovascular, neurologic, or rheumatologic symptoms, for ongoing improvement in the sensitivity and specificity of diagnostic testing, and clarification as to if, when, and how to treat patients with documented Bartonella spp. bacteremia.
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http://dx.doi.org/10.1089/vbz.2018.2353DOI Listing
April 2019

Regional prevalences of Borrelia burgdorferi, Borrelia bissettiae, and Bartonella henselae in Ixodes affinis, Ixodes pacificus and Ixodes scapularis in the USA.

Ticks Tick Borne Dis 2019 02 27;10(2):360-364. Epub 2018 Nov 27.

Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University (NCSU), 1060 William Moore Drive, Raleigh, NC 27607, USA. Electronic address:

The objective of this work was to determine the prevalence of Borrelia and Bartonella species in Ixodes spp. ticks collected from 16 USA states. Genus PCR amplification and sequence analysis of Bartonella and Borrelia 16SsRNA-23SsRNA intergenic regions were performed on DNA extracted from 929 questing adult ticks (671 Ixodes scapularis, 155 Ixodes affinis, and 103 Ixodes pacificus). Overall, 129/929 (13.9%) Ixodes ticks were PCR positive for Borrelia burgdorferi sensu stricto, 48/929 for B. bissettiae whereas 23/929 (2.5%) were PCR positive for a Bartonella henselae. Borrelia bissettiae or B. burgdorferi s.s. and B. henselae co-infections were found in I. affinis from North Carolina at a rate of 4.5%; in a single I. scapularis from Minnesota, but not in I. pacificus. For both bacterial genera, PCR positive rates were highly variable depending on geographic location and tick species, with Ixodes affinis (n = 155) collected from North Carolina, being the tick species with the highest prevalence's for both Borrelia spp. (63.2%) and B. henselae (10.3%). Based on the results of this and other published studies, improved understanding of the enzootic cycle, transmission dynamics, and vector competence of Ixodes species (especially I. affinis) for transmission of Borrelia spp. and B. henselae should be a public health research priority.
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http://dx.doi.org/10.1016/j.ttbdis.2018.11.015DOI Listing
February 2019

Evaluation of cell culture-grown Bartonella antigens in immunofluorescent antibody assays for the serological diagnosis of bartonellosis in dogs.

J Vet Intern Med 2018 Nov 11;32(6):1958-1964. Epub 2018 Oct 11.

Department of Clinical Sciences and the Intracellular Pathogens Research Laboratory, Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina.

Background: Because of poor sensitivity and questionable specificity of immunofluorescent antibody assays (IFAs), serological diagnosis of Bartonella species infections in dogs remains challenging. Despite limitations, IFA testing is the historical "gold standard" for Bartonella serodiagnosis in animals and humans. Because most diagnostic laboratories test against only 1 or 2 Bartonella spp., testing against a broader panel of Bartonella antigens may enhance diagnostic sensitivity and specificity.

Objective: To evaluate the sensitivity and specificity of Bartonella IFA using 8 cell culture-grown Bartonella spp. isolates.

Animals: Archived serum samples from 34 Bartonella spp. naturally exposed, polymerase chain reaction (PCR)-positive dogs and from 26 PCR-negative and IFA-negative dogs.

Methods: Bartonella IFA sensitivity and specificity were assessed using cell culture-grown whole cell antigens derived from 3 Bartonella henselae (Bh) strains (Bh Houston 1, Bh San Antonio Type 2, Bh California 1), 3 Bartonella vinsonii subsp. berkhoffii genotypes (Bvb I, II, and III), Bartonella koehlerae (Bk), and Bartonella quintana (Bq).

Results: Only 62% of 34 Bartonella spp. PCR-positive dogs were seroreactive to any of the 8 Bartonella IFA antigens, indicating low IFA sensitivity. PCR-positive dogs were most often IFA seroreactive to Bq (n = 15), to Bvb II (n = 13), or to both (n = 9) antigens. Of the 26 previously IFA-negative/PCR-negative dogs, 4 (15%) were seroreactive using the expanded antigen panel.

Conclusion And Clinical Importance: Despite IFA testing of dogs against 8 different Bartonella isolates, IFA sensitivity remained poor, and specificity was only 85%. Development of a reliable serological assay is needed to facilitate the diagnosis of Bartonella infection in dogs.
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http://dx.doi.org/10.1111/jvim.15301DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6271329PMC
November 2018

Bartonella quintana and Bartonella vinsonii subsp. vinsonii bloodstream co-infection in a girl from North Carolina, USA.

Med Microbiol Immunol 2019 Feb 24;208(1):101-107. Epub 2018 Sep 24.

Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Dr., Raleigh, NC, 27607, USA.

The genus Bartonella consists of globally distributed and highly diverse alpha-proteobacteria that infect a wide-range of mammals. Medically, Bartonella spp. constitute emerging, vector-borne, zoonotic, intravascular organisms that induce long-lasting bacteremia in reservoir-adapted (passive carrier of a microorganism) hosts. At times, these bacteria are accidentally transmitted by animal scratches, bites, needles sticks or vectors to animal or human hosts. We report the first documented human case of blood stream infection with Bartonella vinsonii subsp. vinsonii in a girl from North Carolina, USA, who was co-infected with Bartonella quintana. Limitations of Bartonella spp. serology and the challenges of microbiological culture and molecular diagnostic confirmation of co-infection with more than one Bartonella spp. are discussed. When and where these infections were acquired is unknown; however, exposure to rodents, fleas and cats in the peri-equestrian environment was a suspected source for transmission of both organisms.
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http://dx.doi.org/10.1007/s00430-018-0563-0DOI Listing
February 2019

Bartonella henselae in small Indian mongooses (Herpestes auropunctatus) from Grenada, West Indies.

Vet Microbiol 2018 Mar 9;216:119-122. Epub 2018 Feb 9.

Anatomy, Physiology, and Pharmacology Academic Program, School of Veterinary Medicine, St. George's University, St. George's, Grenada, West Indies.

Many mammals are established hosts for the vector borne bacterial genus, Bartonella. Small Indian mongooses (Herpestes auropunctatus) have only been reported as a possible host for Bartonella henselae in southern Japan. Confirming Bartonella presence in mongooses from other regions in the world may support their role as potential reservoirs of this human pathogen. Specifically, documenting Bartonella in Caribbean mongooses would identify a potential source of zoonotic risk with mongoose-human contact in the New World. Using serological and molecular techniques, we investigated B. henselae DNA and specific antibody prevalence in 171 mongooses from all six parishes in Grenada, West Indies. Almost a third (32.3%, 54/167) of the tested mongooses were B. henselae seropositive and extracted DNA from 18/51 (35.3%) blood pellets  were PCR positive for the citrate synthase (gltA) and/or the β subunit of RNA polymerase (rpoB) genes. All sequences were identical to B. henselae genotype I, as previously reported from Japan. This study confirms the role of small Indian mongooses as a natural reservoir of B. henselae in the New World.
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http://dx.doi.org/10.1016/j.vetmic.2018.02.009DOI Listing
March 2018

First description of Bartonella koehlerae infection in a Spanish dog with infective endocarditis.

Parasit Vectors 2017 May 19;10(1):247. Epub 2017 May 19.

Hospital Clínic Veterinari, Carrer de l´Hospital, Campus UAB, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.

Background: Bartonella koehlerae has been recently described as a new cat- and cat fleas-associated agent of culture-negative human endocarditis. It has been also encountered in one dog from Israel and six dogs from the USA, but other clinically relevant reports involving this bacterium are lacking.

Results: A 7-year-old intact male mixed dog presented with clinico-pathological signs consistent with mitral endocarditis and cutaneous hemangiosarcoma. Molecular studies revealed the presence of Bartonella koehlerae DNA in samples from blood and mitral valve tissue.

Conclusions: This is the first description of B. koehlerae in Spain, corroborating that it can also be detected in dogs. Bartonella koehlerae infection should also be considered in Spain in humans and dogs presenting with clinical disease suggestive of it, such as culture-negative endocarditis.
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http://dx.doi.org/10.1186/s13071-017-2188-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5437684PMC
May 2017

Detection and prevalence of four different hemotropic Mycoplasma spp. in Eastern North Carolina American black bears (Ursus americanus).

Comp Immunol Microbiol Infect Dis 2017 Feb 23;50:106-109. Epub 2016 Dec 23.

Environmental Medicine Consortium, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Electronic address:

Hemotropic Mycoplasma spp. are globally emerging, obligate parasitic, epierythrocytic bacteria that infect many vertebrates, including humans. Hemoplasma infection can cause acute life-threatening symptoms or lead to a chronic sub-clinical carrier state. Hemotropic Mycoplasma spp. transmission, prevalence, and host specificity are uncertain. The purpose of this study was to determine the molecular prevalence of Mycoplasma species in blood from 68 free-ranging black bears from the eastern coast of North Carolina. DNA amplification of Mycoplasma 16S rRNA gene identified four distinct species infecting 34/68 (50%) of the black bear blood samples, including Candidatus M. haematoparvum. The high prevalence of hemotropic Mycoplasma infection in this wildlife species highlights the importance of understanding intra and inter species transmission. Black bears may play a role in the transmission of hemotropic Mycoplasma spp. between animals, arthropod vectors, and humans. Further studies are needed to elucidate black bears as a potential reservoir for hemotropic Mycoplasma infections.
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http://dx.doi.org/10.1016/j.cimid.2016.12.002DOI Listing
February 2017

Detection of Mycoplasma haemocanis, Mycoplasma haematoparvum, Mycoplasma suis and other vector-borne pathogens in dogs from Córdoba and Santa Fé, Argentina.

Parasit Vectors 2016 12 15;9(1):642. Epub 2016 Dec 15.

Intracellular Pathogens Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 Williams Moore Dr., Raleigh, NC, 27607, USA.

Background: In Argentina, only very few reports are available for canine tick-borne diseases where most are related to parasitic diseases. The objective of this survey was to investigate the prevalence of tick-borne pathogens in 70 dogs from Santa Fé and Córdoba, Argentina.

Methods: Microscopic blood smear examination as well as polymerase chain reaction (PCR) amplification using species-specific markers of Anaplasma, Babesia, Bartonella, Borrelia, Ehrlichia, Francisella, Mycoplasma (hemotropic group) and Rickettsia, followed by DNA sequencing were used to establish the prevalence of each infecting pathogen.

Results: Blood smear analysis showed 81% (57/70) prevalence of structures morphologically compatible with hemotropic mycoplasmas. No structures resembling either piroplasms or Anaplasma/Ehrlichia were detected. Hemotropic mycoplasma species (Mycoplasma haematoparvum, Mycoplasma haemocanis and Mycoplasma suis) were the most prevalent pathogens detected with an overall prevalence of 77.1%. Anaplasma platys was detected and identified in 11 of the 70 dogs (15.7%), meanwhile two Bartonella spp. (B. clarridgeiae and an uncharacterized Bartonella sp.) and Babesia vogeli were detected at 3 and 7% prevalence, respectively.

Conclusions: The work presented here describes a high molecular prevalence for hemotropic mycoplasma species in each of the five locations selected. Three Mycoplasma spp., including Mycoplasma suis, reported for the first time in dogs have been identified by DNA amplification and sequencing. This study highlights the risk that these bacterial pathogens represent for companion animals and, due to their potential zoonotic nature, also for people.
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http://dx.doi.org/10.1186/s13071-016-1920-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5160022PMC
December 2016

Prevalence of Anaplasma phagocytophilum in North Carolina Eastern Black Bears ( Ursus americanus ).

J Wildl Dis 2016 10 1;52(4):968-970. Epub 2016 Aug 1.

1 College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, North Carolina 27607, USA.

We detected Anaplasma phagocytophilum by DNA amplification in whole blood from free-ranging, hunter-killed American black bears ( Ursus americanus ) from the east coast of North Carolina, US. Molecular prevalence for Anaplasma phagocytophilum was 3% from 68 black bears. No DNA of other Anaplasma or Ehrlichia spp. was identified.
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http://dx.doi.org/10.7589/2016-02-036DOI Listing
October 2016

Molecular identification and bioinformatics analysis of a potential anti-vector vaccine candidate, 15-kDa salivary gland protein (Salp15), from Ixodes affinis ticks.

Ticks Tick Borne Dis 2016 Feb 8;7(1):46-53. Epub 2015 Aug 8.

Center for Molecular Medicine, College of Sciences, Old Dominion University, Norfolk, VA 23529, USA; Department of Biological Sciences, Old Dominion University, Norfolk, VA 23529, USA. Electronic address:

Salp15, a 15-kDa salivary gland protein plays an important role in tick blood-feeding and transmission of Borrelia burgdorferi, the causative agent of Lyme borreliosis. The comparative studies reveal that Salp15 is a genetically conserved protein across various Ixodes species. In this study, we have identified a Salp15 homolog, designated as Iaff15, from Ixodes affinis ticks that are the principal enzootic vectors of B. burgdorferi sensu stricto in the southeastern part of the United States. Comparison of the annotated amino acid sequences showed that Iaff15 share 81% homology with I. sinensis Salp15 homolog and 64% homology with I. scapularis Salp15. Phylogenetic analysis revealed that Iaff15 come within the same clade with I. sinensis, I. scapularis, and I. pacificus Salp15 homologs. The bioinformatics analysis of the posttranslational modifications prediction revealed that all the Salp15 family members contain glycosylation sites. In addition, Iaff15 carried a higher number of Casein Kinase II phosphorylation sites in comparison to the other Salp15 family members. Collectively, high sequence conservation distributed over the entire amino acids sequence not only suggests an important role for Iaff15 in I. affinis blood feeding and vector-pathogen interactions but may also lead to the development of an anti-vector vaccine against this group of ticks.
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http://dx.doi.org/10.1016/j.ttbdis.2015.08.003DOI Listing
February 2016

Infection with Bartonella henselae in a Danish family.

J Clin Microbiol 2015 May 4;53(5):1556-61. Epub 2015 Mar 4.

Intracellular Pathogens Research Laboratory, Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA

Bartonella species constitute emerging, vector-borne, intravascular pathogens that produce long-lasting bacteremia in reservoir-adapted (natural host or passive carrier of a microorganism) and opportunistic hosts. With the advent of more sensitive and specific diagnostic tests, there is evolving microbiological evidence supporting concurrent infection with one or more Bartonella spp. in more than one family member; however, the mode(s) of transmission to or among family members remains unclear. In this study, we provide molecular microbiological evidence of Bartonella henselae genotype San Antonio 2 (SA2) infection in four of six Danish family members, including a child who died of unknown causes at 14 months of age.
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http://dx.doi.org/10.1128/JCM.02974-14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4400761PMC
May 2015

Potentially novel Ehrlichia species in horses, Nicaragua.

Emerg Infect Dis 2015 Feb;21(2):335-8

Ehrlichia sp. DNA was amplified from 4 Ehrlichia-seroreactive horses from Mérida, Nicaragua. Sequencing of 16S rDNA, sodB, and groEL genes indicated that the bacterium is most likely a novel Ehrlichia species. The tick vector and the potential for canine and human infection remain unknown.
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http://dx.doi.org/10.3201/eid2102.140290DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4313632PMC
February 2015

Vector-borne pathogens in arctic foxes, Vulpes lagopus, from Canada.

Res Vet Sci 2015 Apr 16;99:58-9. Epub 2014 Dec 16.

Intracellular Pathogens Research Laboratory (IPRL), Center for Comparative Medicine and Translational Research College of Veterinary Medicine, North Carolina State University, NC, USA. Electronic address:

Because of the relatively low biodiversity within arctic ecosystems, arctic foxes, Vulpes lagopus, could serve as sentinels for the study of changes in the ecology of vector-borne zoonotic pathogens. The objective of this study was to determine the molecular prevalence of 5 different genera of vector borne pathogens (Anaplasma, Babesia, Bartonella, Ehrlichia, and Hemotropic Mycoplasma spp.) using blood collected from 28 live-trapped arctic foxes from the region of Karrak Lake, Nunavut, Canada. Bartonella henselae (n = 3), Mycoplasma haemocanis (n = 1), Ehrlichia canis (n = 1), and an Anaplasma sp. (n = 1) DNA were PCR amplified and subsequently identified by sequencing. This study provides preliminary evidence that vector borne pathogens, not typically associated with the arctic ecosystem, exist at low levels in this arctic fox population, and that vector exposure, pathogen transmission dynamics, and changes in the geographic distribution of pathogens over time should be investigated in future studies.
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http://dx.doi.org/10.1016/j.rvsc.2014.12.011DOI Listing
April 2015

Bartonella henselae infections in an owner and two Papillon dogs exposed to tropical rat mites (Ornithonyssus bacoti).

Vector Borne Zoonotic Dis 2014 Oct;14(10):703-9

Intracellular Pathogens Research Laboratory, Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University , Raleigh, North Carolina.

After raccoons were trapped and removed from under a house in New York, the owner and her two Papillon dogs became infested with numerous rat mites (Ornithonyssus bacoti). Two weeks later, both dogs developed pruritus, progressively severe vesicular lesions, focal areas of skin exfoliation, swelling of the vulva or prepuce, abdominal pain, and behavioral changes. Two months after the mite infestation, the owner was hospitalized because of lethargy, fatigue, uncontrollable panic attacks, depression, headaches, chills, swollen neck lymph nodes, and vesicular lesions at the mite bite sites. Due to ongoing illness, 3 months after the mite infestation, alcohol-stored mites and blood and serum from both dogs and the owner were submitted for Bartonella serology and Bartonella alpha Proteobacteria growth medium (BAPGM) enrichment blood culture/PCR. Bartonella henselae DNA was amplified and sequenced from blood or culture specimens derived from both dogs, the owner, and pooled rat mites. Following repeated treatments with doxycycline, both dogs eventually became B. henselae seronegative and blood culture negative and clinical signs resolved. In contrast, the woman was never B. henselae seroreactive, but was again PCR positive for B. henselae 20 months after the mite infestation, despite prior treatment with doxycycline. Clinicians and vector biologists should consider the possibility that rat mites may play a role in Bartonella spp. transmission.
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http://dx.doi.org/10.1089/vbz.2013.1492DOI Listing
October 2014

Bilateral mandibular pyogranulomatous lymphadenitis and pulmonary nodules in a dog with Bartonella henselae bacteremia.

Can Vet J 2014 Oct;55(10):970-4

Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman, Washington 99164-7060, USA (MD Tucker, Sellon, RL Tucker); IDEXX Reference Laboratories Inc., Pullman, Washington, USA (Wills); Dillon Small Animal Hospital, Dillon, Montana, USA (Simonsen); Intracellular Pathogens Research Laboratory, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA (Maggi, Breitschwerdt).

This report describes a 2-year-old collie dog with pulmonary nodules, visualized by computed tomographic (CT) scan, with evidence of Bartonella henselae bacteremia and pyogranulomatous lymphadenitis. Clinical signs resolved with antimicrobial therapy.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4187381PMC
October 2014

Detection of Bartonella species in the blood of veterinarians and veterinary technicians: a newly recognized occupational hazard?

Vector Borne Zoonotic Dis 2014 Aug;14(8):563-70

1 Departments of Internal Medicine and Pediatrics, Duke University School of Medicine , Durham, North Carolina.

Background: Bartonella species are important emerging pathogens in human and veterinary medicine. In the context of their daily activities, veterinary professionals have frequent animal contact and arthropod exposures. Detection of Bartonella spp. using traditional culture methods has been limited by poor sensitivity, making it difficult to determine the prevalence of infection in this population. We have developed a detection method combining enrichment culture and molecular amplification, which increases testing sensitivity.

Methods: We performed a cross-sectional study to determine the prevalence of detectable Bartonella spp. in the blood of veterinary personnel and nonveterinary control subjects. Bartonella was detected by enrichment blood culture with conventional PCR followed by DNA sequencing. RESULTS were correlated with epidemiological variables and symptoms.

Results: We detected DNA from at least one Bartonella species in 32 (28%) of the 114 veterinary subjects. After DNA sequencing, the Bartonella species could be determined for 27 of the 32 infected subjects, including B. henselae in 15 (56%), B. vinsonii subsp. berkhoffii in seven (26%), B. koehlerae in six (22%), and a B. volans-like sequence in one (4%). Seventy percent of Bartonella-positive subjects described headache compared with 40% of uninfected veterinarians (p=0.009). Irritability was also reported more commonly by infected subjects (68% vs. 43%, p=0.04).

Conclusions: Our study supports an emerging body of evidence that cryptic Bartonella bloodstream infection may be more frequent in humans than previously recognized and may induce symptoms. Longitudinal studies are needed to determine the natural course and clinical features of Bartonella infection.
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http://dx.doi.org/10.1089/vbz.2013.1512DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4117269PMC
August 2014

Intravascular persistence of Anaplasma platys, Ehrlichia chaffeensis, and Ehrlichia ewingii DNA in the blood of a dog and two family members.

Parasit Vectors 2014 Jul 1;7:298. Epub 2014 Jul 1.

Intracellular Pathogens Research Laboratory and the Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA.

Background: Anaplasmosis, caused by Anaplasma phagocytophilum and Anaplasma platys, and ehrlichiosis, caused by Ehrlichia chaffeensis, Ehrlichia ewingii, the "Panola Mountain Ehrlichia" and Ehrlichia muris-like pathogens have been identified as emerging tick borne infectious diseases in dogs and human patients. Persistent intravascular infection with these bacteria is well documented in dogs, but is less well documented in human beings.

Methods: Serology and PCR targeting multiple microbial genes, followed by DNA sequencing, was used to test sequential blood samples. Tissue culture isolation was attempted in two laboratories.

Results: A. platys, E. chaffeensis, and E. ewingii DNA was amplified from two Anaplasma and Ehrlichia seronegative family members and their dog, all lacking typical symptoms of anaplasmosis or ehrlichiosis. Following treatment with doxycycline, the dog and mother were Anaplasma and Ehrlichia spp. PCR negative.

Conclusions: Sequential PCR testing provided molecular evidence supporting intravascular persistence of A. platys and Ehrlichia spp. in two humans and their dog. Diagnosticians and clinicians should consider the potential for co-infections due to these tick borne organisms.
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http://dx.doi.org/10.1186/1756-3305-7-298DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4089936PMC
July 2014
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