Bartonella in Ticks

Bartonella In Ticks- Medical Abstracts

Bartonella & Ticks

Arch Neurol. 2001 Sep;58(9):1357-63. Concurrent infection of the central nervous system by Borrelia burgdorferi and Bartonella henselae: evidence for a novel tick-borne disease complex. Eskow E, Rao RV, Mordechai E. Hunterdon Medical Center, Flemington, NJ, USA.

OBJECTIVES: To investigate Bartonella henselae as a potential human tick-borne pathogen and to evaluate its role as a coinfecting agent of the central nervous system in the presence of neuroborreliosis. DESIGN: Case report study. SETTING: A primary health care center in Flemington, NJ, and the Department of Research and Development at Medical Diagnostic Laboratories LLC in Mt Laurel, NJ. SUBJECTS: Two male patients (aged 14 and 36 years) and 2 female patients (aged 15 and 30 years, respectively) with a history of tick bites and Lyme disease. MAIN OUTCOME MEASURES: Laboratory and diagnostic findings before and after antimicrobial therapy. RESULTS: Patients residing in a Lyme-endemic area of New Jersey with ongoing symptoms attributed to chronic Lyme disease were evaluated for possible coinfection with Bartonella species. Elevated levels of B henselae-specific antibodies were found in these patients using the immunofluorescent assay. Bartonella henselae-specific DNA was detected in their blood. None of these patients exhibited the clinical characteristics of cat-scratch disease. Findings of cerebrospinal fluid analysis revealed the presence of both B henselae- and Borrelia burgdorferi-specific DNA. Bartonella henselae-specific DNA was also detected in live deer ticks obtained from the households of 2 of these patients. CONCLUSIONS: Our data implicate B henselae as a potential human tick-borne pathogen. Patients with a history of neuroborreliosis who have incomplete resolution of symptoms should be evaluated for B henselae infection.Publication Types:* Case Reports PMID: 11559306 [PubMed - indexed for MEDLINE]

Med Parazitol (Mosk). 2005 Apr-Jun;(2):44-8. [Bartonellosis and a possible role of Ixodes ticks (family Ixodidae, order Parasitiformes) in the transmission of pathogenic Bartonella bacteria] [Article in Russian]

The papers reviews the literature on bartonellosis and a role of Ixodes ticks, including the representatives of the genus Ixodes, in the circulation and transmission of Bartonella bacteria. It shows that man can be infected with pathogenic Bartonella bacteria by the bite of ticks. The paper also presents data on tick-transmitted human and animal mixed infections, including bartonellosis.PMID: 15984622 [PubMed - in process]

Comp Immunol Microbiol Infect Dis. 2002 Jul;25(4):229-36. Investigation of Bartonella infection in ixodid ticks from California. Chang CC, Hayashidani H, Pusterla N, et al Department of Public Health, Institute of Environmental Health, China Medical College, Taichung, Taiwan, ROC.

A total of 1253 ixodid ticks (254 tick pools) collected between the end of 1995 and the spring of 1997 from six California counties (El Dorado, Los Angeles, Orange, Santa Cruz, Shasta and Sonoma) were examined for the presence of Bartonella DNA by PCR of the citrate synthase gene. Of 1,119 adult Ixodes pacificus ticks tested, 26 (11.6%) of 224 pools, each containing five ticks, were positive (minimum percentage of ticks harboring detectable Bartonella DNA, 2.3%). Bartonella PCR-positive ticks were identified in five counties but none of the ticks from Los Angeles County was positive. Among 47 nymphal I. pacificus ticks collected in Sonoma County, one (10%) positive pool out of 10 pools was identified (minimum percentage of ticks harboring detectable Bartonella DNA, 2.1%). Among the 54 Dermacentor occidentalis grouped in 12 pools from Orange County, one pool (8.3%) was PCR positive for Bartonella and similarly one pool (14.3%) was positive among the 30 Dermacentor variabilis ticks grouped in seven pools. None of the three D. occidentalis from El Dorado County were positive. None of the nine tick pools positive for Ehrlichia phagocytophila were positive for Bartonella. Following our previous findings of Bartonella PCR-positive adult I. pacificus ticks in central coastal California, this is the first preliminary report of the presence of Bartonella DNA in I. pacificus nymphs and in Dermacentor sp. ticks. Distribution of Bartonella among ixodid ticks appears widespread in California. PMID: 12135237 [PubMed - indexed for MEDLINE]

J Clin Microbiol. 1999 Jul;37(7):2215-22. Detection and identification of Ehrlichia, Borrelia burgdorferi sensu lato, and Bartonella species in Dutch Ixodes ricinus ticks. Schouls LM, Van De Pol I, Rijpkema SG, et al. Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, Bilthoven, The Netherlands.

A sensitive and specific PCR hybridization assay was developed for the simultaneous detection and identification of Ehrlichia and Borrelia burgdorferi sensu lato. In separate assays the 16S rRNA gene of Ehrlichia species and the 23S-5S rRNA spacer region of B. burgdorferi sensu lato were amplified and labeled by PCR. These PCR products were used in a reverse line blot hybridization assay in which oligonucleotide probes are covalently linked to a membrane in parallel lines. Hybridization of the samples with the oligonucleotide probes on this membrane enabled the simultaneous detection and identification of Ehrlichia, B. burgdorferi, and Bartonella species in 40 different samples. The application of the assay to DNA extracts from 121 Ixodes ricinus ticks collected from roe deer demonstrated that 45% of these ticks carried Ehrlichia DNA. More than half of these positive ticks carried species with 16S rRNA gene sequences closely related to those of E. phagocytophila and the human granulocytic ehrlichiosis agent. The majority of the other positive ticks were infected with a newly identified Ehrlichia-like species. In addition, 13% of the ticks were infected with one or more B. burgdorferi genospecies. In more than 70% of the ticks 16S rRNA gene sequences for Bartonella species or other species closely related to Bartonella were found. In five of the ticks both Ehrlichia and B. burgdorferi species were detected. PMID: 10364588 [PubMed - indexed for MEDLINE]

J Med Entomol. 2005 May;42(3):473-80. Detection of Rickettsia, Borrelia, and Bartonella in Carios kelleyi (Acari: Argasidae). Loftis AD, Gill JS, Schriefer ME, et al Viral and Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.

Carios kelleyi (Colley & Kohls 1941), a tick associated with bats and bat habitats, has been reported to feed on humans, but there is little published data regarding the presence of vector-borne pathogens in these ticks. C. kelleyi nymphs and adults were collected from residential and community buildings in Jackson County, Iowa, and tested by polymerase chain reaction for Rickettsia, Borrelia, Bartonella, Coxiella, and Anaplasma. Rickettsia DNA was detected in 28 of 31 live ticks. Sequences of the 17-kDa and rOmpA genes suggest that this agent is a novel spotted fever group Rickettsia. Transstadial and transovarial transmission of this Rickettsia were demonstrated. The flagellin gene of a Borrelia, closely related to B. turicatae, was detected in one of 31 live ticks. The 16S-23S intergenic spacer region of Bartonella henselae also was detected in one of 31 live ticks. Coxiella or A. phagocytophilum DNA were not detected in these ticks. PMID: 15962801 [PubMed - in process]

Vector Borne Zoonotic Dis. 2004 Winter;4(4):306-9. Semi-nested PCR detection of Bartonella henselae in Ixodes persulcatus ticks from Western Siberia, Russia. Morozova, Cabello, Dobrotvorsky Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia.

Questing adult Ixodes persulcatus ticks from Western Siberia, Russia were tested for infections with Bartonella spp. using seminested PCR assay with primers specific to the groEL gene. The proportion of ticks infected with Bartonella spp. was 44% in 2002 (n = 50) and 38% in 2003 (n = 50). Nucleotide sequences of a portion of the PCR products corresponded to Bartonella henselae species. PMID: 15671737 [PubMed - indexed for MEDLINE]

J Infect Dis. 2005 Feb 15;191(4):607-11. Epub 2005 Jan 10. Bartonella quintana in a 4000-year-old human tooth. Drancourt M, Tran-Hung L, Courtin J, Lumley H, Raoult D Unite des Rickettsies, CNR UMR 6020, IFR 48, Marseille, France.

Bacteria of the genus Bartonella are transmitted by ectoparasites (lice, fleas, ticks) and have mammalian reservoirs in which they cause chronic, asymptomatic bacteremia. Humans are the reservoir of B. quintana, the louse-borne agent of trench fever. We detected DNA of B. quintana in the dental pulp of a person who died 4000 years ago. PMID: 15655785 [PubMed - indexed for MEDLINE]

Vet Res. 2005 Jan-Feb;36(1):79-87. Evidence of Bartonella sp. in questing adult and nymphal Ixodes ricinus ticks from France and co-infection with Borrelia burgdorferi sensu lato and Babesia sp. Halos L, Jamal T, et al. UMR 956 INRA/AFSSA/ENVA/UVPM, Microbiologie, Ecole Nationale Veterinaire, 7 avenue du General de Gaulle, 94 700 Maisons-Alfort, France.

Ticks are known vectors for a wide range of pathogenic microorganisms. Their role in the transmission of some others is so far only suspected. Ticks can transmit multiple pathogens, however, little is known about the co-existence of these pathogens within questing ticks. We looked for the presence of DNA from three micro-organisms, Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. which are known or suspected tick-borne pathogens, using a cohort of 92 questing Ixodes ricinus ticks collected from pastures in northern France. DNA was extracted from each individual tick and the presence of the three pathogens was investigated using Polymerase Chain Reaction (PCR) amplification. Nine among 92 samples (9.8%) demonstrated PCR products using Bartonella specific primers, 3 among 92 (3.3%) using Borrelia burgdorferi sensu lato specific primers and 19 among 92 (20.6%) using Babesia specific primers. Seven among 92 samples (7.6%) were PCR positive for at least two of the pathogens and one sample was positive for all three. Adult ticks (12/18; 67%) showed significantly higher infection rates compared to nymphs (11/74; 15%) for all three pathogens (P < 0.001). This study is the demonstration of the simultaneous presence of Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. in questing Ixodes ricinus ticks. PMID: 15610725 [PubMed - indexed for MEDLINE]

J Parasitol. 2004 Jun;90(3):485-9. Ectoparasites of gray squirrels in two different habitats and screening of selected ectoparasites for bartonellae. Durden LA, Ellis BA, et al. Department of Biology and Institute of Arthropodology and Parasitology, Georgia Southern University, Statesboro, Georgia 30460-8042, USA.

Gray squirrels, Sciurus carolinensis, were livetrapped in 2 different habitat types, woodland (67 squirrels) and parkland (53 squirrels), in southeastern Georgia. Ectoparasites were recovered from anesthetized squirrels and compared between hosts from the 2 habitats. Because of the absence of low vegetation in parkland habitats, it was hypothesized that the ectoparasite fauna, especially ticks and chiggers, would be more diverse on woodland squirrels. The results were generally in agreement with this hypothesis. Seventeen species of ectoparasites were recovered from woodland squirrels, compared with 6 species from parkland squirrels. Five species of ticks and 3 species of chiggers parasitized the woodland squirrels compared with no ticks or chiggers on the parkland squirrels. Significantly higher infestation prevalences were recorded on woodland compared with parkland squirrels for the flea Orchopeas howardi, the tick Amblyomma americanum, and the mesostigmatid mite Androlaelaps fahrenholzi. The mean intensity for O. howardi also was significantly higher on woodland than on parkland squirrels. Because a new strain of Bartonella sp. was isolated recently from S. carolinensis in Georgia, selected ectoparasites from this study were screened for bartonellae by polymerase chain reaction (PCR). Some of the fleas and lice, but none of the mites tested, were PCR positive, suggesting that fleas, or lice, or both, might be vectors of bartonellae between squirrels. Six distinct strains of Bartonella sp. were detected, 2 in fleas and 4 in lice. PMID: 15270090 [PubMed - indexed for MEDLINE]

J Clin Microbiol. 2004 Jun;42(6):2799-801. Prevalence of Borrelia burgdorferi, Bartonella spp., Babesia microti, and Anaplasma phagocytophila in Ixodes scapularis ticks collected in Northern New Jersey. Adelson ME, Rao RV, et al. Medical Diagnostic Laboratories L.L.C., 133 Gaither Dr., Suite C, Mt. Laurel, NJ 08054, USA.

PCR analysis of Ixodes scapularis ticks collected in New Jersey identified infections with Borrelia burgdorferi (33.6%), Babesia microti (8.4%), Anaplasma phagocytophila (1.9%), and Bartonella spp. (34.5%). The I. scapularis tick is a potential pathogen vector that can cause coinfection and contribute to the variety of clinical responses noted in some tick-borne disease patients. PMID: 15184475 [PubMed - indexed for MEDLINE]

Vector Borne Zoonotic Dis. 2002 Winter;2(4):265-73. Epidemiology and impact of coinfections acquired from Ixodes ticks. Belongia EA Epidemiology Research Center, Marshfield Clinic Research Foundation, Marshfield, Wisconsin 54449, USA.

Ixodes scapularis and other ticks in the Ixodes ricinus complex may transmit multiple pathogens, but research on coinfections has been limited. Coinfections occur with varying frequency in ticks, but single infections are more common than dual infections. The proportion of I. scapularis or I. ricinus ticks coinfected with both Borrelia burgdorferi sensu lato and Anaplasma phagocytophila is generally low, ranging from < 1% to 6% in six geographic areas. A higher prevalence of tick coinfection (26%) has been reported in Westchester County, New York. Genetic variants of the human disease-causing strain of A. phagocytophila are present in some tick populations, and they may affect the risk of coinfection or clinical illness. The proportion of Ixodes ticks coinfected with B. burgdorferi and Babesia microti has ranged from 2% in New Jersey to 19% on Nantucket Island, Massachusetts. In humans, cross-sectional seroprevalence studies have found markers of dual infection in 9-26% of patients with a tick-borne infection, but such studies often fail to distinguish simultaneous coinfection from sequential infections. Several studies have prospectively assessed the occurrence of acute coinfection. Among patients with a confirmed tick-borne infection, coinfection rates as high as 39% have been reported. The most commonly recognized coinfection in most of the eastern United States is Lyme borreliosis (LB) and babesiosis, accounting for approximately 80% of coinfections. LB and human granulocytic ehrlichiosis coinfections are less common, occurring in 3-15% of patients with a tick-borne infection in Connecticut or Wisconsin. Studies of clinical outcomes suggest that patients with acute Babesia coinfection have more severe symptoms and a longer duration of illness than patients with LB alone, but the risk of spirochete dissemination is similar. Coinfections can modify the immune response and alter the severity of arthritis in animal models. Future coinfection research should focus on long-term clinical outcomes, the role of genetic variants, immunologic effects, and the potential role of Bartonella species as tick-borne pathogens. PMID: 12804168 [PubMed - indexed for MEDLINE]

Emerg Infect Dis. 2003 Mar;9(3):329-32. Bartonella henselae in Ixodes ricinus ticks (Acari: Ixodida) removed from humans, Belluno province, Italy. Sanogo YO, Zeaiter Z, Caruso G, et al Faculte de Medecine, Marseille, France.

The potential role of ticks as vectors of Bartonella species has recently been suggested. In this study, we investigated the presence of Bartonella species in 271 ticks removed from humans in Belluno Province, Italy. By using primers derived from the 60-kDa heat shock protein gene sequences, Bartonella DNA was amplified and sequenced from four Ixodes ricinus ticks (1.48%). To confirm this finding, we performed amplification and partial sequencing of the pap31 protein and the cell division protein ftsZ encoding genes. This process allowed us to definitively identify B. henselae (genotype Houston-1) DNA in the four ticks. Detection of B. henselae in these ticks might represent a highly sensitive form of xenodiagnosis. B. henselae is the first human-infecting Bartonella identified from Ixodes ricinus, a common European tick and the vector of various tickborne pathogens. The role of ticks in the transmission of bartonellosis should be further investigated. PMID: 12643827 [PubMed - indexed for MEDLINE]

J Microbiol Methods. 2003 Feb;52(2):251-60. Application of broad-range 16S rRNA PCR amplification and DGGE fingerprinting for detection of tick-infecting bacteria. Schabereiter-Gurtner C, Lubitz W, Rolleke S. Institute of Microbiology and Genetics, University of Vienna, Dr Bohr-Gasse 9, 1030 Vienna, Austria.

Ticks play an important role in the transmission of arthropod-borne diseases of viral, protozoal and bacterial origin. The present article describes a molecular-biological based method, which facilitated the broad-range analyses of bacterial communities in ixodid ticks (Ixodes ricinus). DNA was extracted both from single ticks and pooled adult ticks. Eubacterial 16S rRNA gene fragments (16S rDNA) were amplified by polymerase chain reaction (PCR) with broad-range ribosomal primers. Sequences spanning the hypervariable V3 region of the 16S rDNA and representing individual bacterial taxons were separated by denaturing gradient gel electrophoresis (DGGE). For phylogenetic identification, DGGE bands were exised, cloned and sequenced. In addition, we set up a 16S rDNA clone library which was screened by DGGE. Sequences were compared with sequences of known bacteria listed in the GenBank database. A number of bacteria were affiliated with the genera Rickettsia, Bartonella, and Borrelia, which are known to be pathogenic and transmitted by ticks. Two sequences were related to the yet to be cultivated Haemobartonella. To our knowledge, Haemobartonella has never been directly detected in I. ricinus. In addition, members of the genera Staphylococcus, Rhodococcus, Pseudomonas, and Moraxella were detected, which have not been identified in ticks so far. Two bacteria were most closely related to a rickettsial endosymbiont of an Acanthamoeba sp., and to an endosymbiont (Legionellaceae, Coxiella group) of the microarthropod Folsomia candida. The results prove that 16S rDNA genotyping in combination with DGGE analysis is a promising approach for the detection and identification of bacteria infecting ticks, regardless of whether these bacteria are fastidious, obligate intracellular or noncultivable.

PMID: 12459246 [PubMed - indexed for MEDLINE]

Am J Trop Med Hyg. 2002 Aug;67(2):135-6. First molecular evidence of new Bartonella spp. in fleas and a tick from Peru. Parola P, Shpynov S, Montoya M, et al. Unite des Rickettsies, Faculte de Medecine, Universite de la Mediterranee, Marseille, France.

Fleas, lice, and ticks collected in Peru in a suburban area of Cusco in November 1998 were tested by polymerase chain reaction for the presence of Bartonella DNA using primers amplifying a fragment of the intergenic spacer region (ITS) gene. Three new Bartonella genotypes were detected in Pulex fleas self-collected from the beds and clothes of schoolchildren and adults. A fourth new genotype was also detected from a tick found on a sheep in the same area. One of the genotypes is closely related to B. vinsoni subsp. berkhoffii, and the others seem to originate from unknown Bartonella species, whose medical importance has yet to be clarified. PMID: 12389935 [PubMed - indexed for MEDLINE]

Am J Trop Med Hyg. 2002 Jan;66(1):80-5. Fourteen-year seroepidemiological study of zoonoses in a Greek village. Antoniou M, Economou I, Wang X, et al Laboratory of Clinical Bacteriology Parasitotlogy Zoonoses and Geographical Medicine, Faculty of Medicine, University of Crete, Heraklion, Greece.

A seroepidemiological study carried out in a high-risk village in Crete in 1985-1987 and 1998 showed that although the awareness of the people concerning zoonoses had increased during this period, the situation did not improve: there was a significant increase of the spread of seroprevalence in time and space of Coxiella burnetii, Rickettsia typhi, Brucella sp., and Entamoeba histolytica. Toxoplasma gondii, Rickettsia conorii, Borrelia burgdorferi, Echinococcus granulosus, Leishmania sp., and Fasciola hepatica stayed at the same levels. This first study of Bartonella henselae in Crete showed that 15.9% of the children tested were seropositive. The results indicate that reservoirs and vectors of the pathogens studied are widespread in the environment, and the way of life of the people favors contact with them. Seven of 30 milk samples were positive for Brucella sp. by seminested polymerase chain reaction. PMID: 12135274 [PubMed - indexed for MEDLINE]

J Am Vet Med Assoc. 2001 Apr 1;218(7):1092-7. Assessing the association between the geographic distribution of deer ticks and seropositivity rates to various tick-transmitted disease organisms in dogs. Hinrichsen VL, Whitworth UG, Breitschwerdt EB, et al. Center for Vector-Borne Disease, University of Rhode Island, Kingston 02881, USA.

OBJECTIVE: To determine whether the geographic distribution of deer ticks (Ixodes scapularis) was associated with the distribution of dogs seropositive for various tick-transmitted disease organisms (ie, Borrelia burgdorferi, Rickettsia rickettsii, the human granulocytic ehrlichiosis [HGE] agent, Ehrlichia canis, and Bartonella vinsonii subsp berkhoffii). DESIGN: Serologic survey. SAMPLE POPULATION: Serum samples from 277 dogs in animal shelters and veterinary hospitals in Rhode Island. RESULTS: Overall, 143 (52%) dogs were seropositive for B burgdorferi, 59 (21.3%) were seropositive for R rickettsii, 40 (14.4%) were seropositive for the HGE agent, 8 (2.9%) were seropositive for E canis, and 6 (2.2%) were seropositive for B vinsonii. Regression analysis indicated that the natural logarithm of nymphal deer tick abundance was correlated with rate of seropositivity to the HGE agent and to B burgdorferi but not to rate of seropositivity to R rickettsii, E canis, or B vinsonii. Percentages of samples seropositive for B burgdorferi, R rickettsii, the HGE agent, and E canis were significantly higher for samples from the southwestern part of the state where ticks in general and deer ticks in particular are abundant than for samples from the northern and eastern portions of the state, where ticks are relatively rare. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that all 5 disease agents are in Rhode Island and pose a risk to dogs and humans. Knowledge concerning tick distributions may be useful in predicting the pattern of disease associated with particular tick species and may aid diagnostic, prevention, and control efforts.

PMID: 11318358 [PubMed - indexed for MEDLINE]

J Clin Microbiol. 2001 Apr;39(4):1221-6. Molecular evidence of Bartonella spp. in questing adult Ixodes pacificus ticks in California. Chang CC, Chomel BB, Kasten RW, et al. Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, California 95616, USA.

Ticks are the vectors of many zoonotic diseases in the United States, including Lyme disease, human monocytic and granulocytic ehrlichioses, and Rocky Mountain spotted fever. Most known Bartonella species are arthropod borne. Therefore, it is important to determine if some Bartonella species, which are emerging pathogens, could be carried or transmitted by ticks. In this study, adult Ixodes pacificus ticks were collected by flagging vegetation in three sites in Santa Clara County, Calif. PCR-restriction fragment length polymorphism and partial sequencing of 273 bp of the gltA gene were applied for Bartonella identification. Twenty-nine (19.2%) of 151 individually tested ticks were PCR positive for Bartonella. Male ticks were more likely to be infected with Bartonella than female ticks (26 versus 12%, P = 0.05). None of the nine ticks collected at Baird Ranch was PCR positive for Bartonella. However, 7 (50%) of 14 ticks from Red Fern Ranch and 22 (17%) of 128 ticks from the Windy Hill Open Space Reserve were infected with Bartonella. In these infected ticks, molecular analysis showed a variety of Bartonella strains, which were closely related to a cattle Bartonella strain and to several known human-pathogenic Bartonella species and subspecies: Bartonella henselae, B. quintana, B. washoensis, and B. vinsonii subsp. berkhoffii. These findings indicate that I. pacificus ticks may play an important role in Bartonella transmission among animals and humans. PMID: 11283031 [PubMed - indexed for MEDLINE]

Am J Vet Res. 1997 May;58(5):467-71. Epidemiologic evaluation of the risk factors associated with exposure and seroreactivity to Bartonella vinsonii in dogs. Pappalardo BL, Correa MT, York CC, et al. Department of Companion Animal and Special Species Medicine, College of Veterinary Medicine, North Carolina State University, Raleigh 27606, USA.

OBJECTIVES: To determine seroprevalence to Bartonella vinsonii subsp berkhoffii in a population of sick dogs from North Carolina and Virginia and to evaluate potential risk factors associated with increased likelihood of exposure to the organism. SAMPLE POPULATION: Serum samples from 1,920 sick dogs. PROCEDURE: An indirect fluorescent antibody assay was performed on each sample, and the end-point antibody titer was recorded. A case (seropositive) was defined as a dog with reciprocal titer > or = 64, and a control (seronegative) was defined as a dog with reciprocal titer < 16 that was referred within 0 to 3 days of referral of a corresponding case. From this population, 207 dogs (69 cases and 138 controls) were included in a case-control seroepidemiologic study. RESULTS: 3.6% (69/1,920) of the dogs were seropositive to B vinsonii subsp berkhoffii. Results of the case-control study indicated that seropositive dogs were more likely to live in rural environments, frequently on a farm, were free to roam the neighborhood, and were considered to be predominantly outdoor dogs. Moreover, seropositive dogs were 14 times more likely to have a history of heavy tick exposure. After analysis of the case-control study, a more detailed examination of banked sera from dogs with known tick exposure was performed. High correlation was found between sero-reactivity to B vinsonii and seroreactivity to E canis or B canis (36.0 and 57.1%, respectively). Sera derived from dogs experimentally infected with E canis or R rickettsii did not cross react with B vinsonii antigen. CONCLUSIONS AND CLINICAL RELEVANCE: Several potential risk factors are associated with canine exposure to B vinsonii. Rhipicephalus sanguineus, the tick vector for E canis and B canis, may be involved in B vinsonii transmission among dogs. PMID: 9140552 [PubMed - indexed for MEDLINE]

Rocz Akad Med Bialymst. 1996;41(1):129-35. Unknown species of rickettsiae isolated from Ixodes ricinus tick in Walcz. Kruszewska D, Tylewska-Wierzbanowska S. National Institute of Hygiene, Warsaw, Poland.

Ticks Ixodes ricinus have been harvested in park down-town Walcz in June 1994. Presence of Borrelia burgdorferi and rickettsiae has been tested with PCR. B. burgdorferi and Coxiella burnetii have not been detected in any tick whereas characteristic for other rickettsiae DNA fragment of gene encoding citrate-synthase has been found. Bacterial strain has been cultured from the tick. Biochemical properties of isolated strain has strongly suggested that these bacteria belong to genus Bartonella (Rochalimaea). Further identification of bacterial DNA with RFLP-PCR (restriction fragment length polymorphism-PCR) has shown characteristics of Bartonella bacilliformis species not recognised in Poland until recently. PMID: 8673798 [PubMed - indexed for MEDLINE]

J Clin Microbiol. 2004 June; 42(6): 2799–2801. doi: 10.1128/JCM.42.6.2799-2801 2004, American Society for Microbiology. Prevalence of Borrelia burgdorferi, Bartonella spp., Babesia microti, and Anaplasma phagocytophila in Ixodes scapularis Ticks Collected in Northern New Jersey. Martin E. Adelson,1 Raja-Venkitesh S. Rao,1 Richard C. Tilton,1 Kimberly Cabets,1 Eugene Eskow,2 Lesley Fein,3 James L. Occi,4 and Eli Mordechai1*

Medical Diagnostic Laboratories L.L.C., Mt. Laurel, New Jersey 08504,1 4 Walter Foran Boulevard, Suite 103, Flemington, New Jersey 08822,2 1099 Bloomfield Avenue, West Caldwell, New Jersey, 07006,3 Graduate Program in Biology, Rutgers University, New Jersey 071024 *Corresponding author. Mailing address: Medical Diagnostic Laboratories L.L.C., 133 Gaither Dr., Suite C, Mt. Laurel, NJ 08054. Phone: (856) 608-1696. Fax: (856) 608-1667. E-mail: Received September 15, 2003; Revised January 7, 2004; Accepted March 9, 2004.

PCR analysis of Ixodes scapularis ticks collected in New Jersey identified infections with Borrelia burgdorferi (33.6%), Babesia microti (8.4%), Anaplasma phagocytophila (1.9%), and Bartonella spp. (34.5%). The I. scapularis tick is a potential pathogen vector that can cause coinfection and contribute to the variety of clinical responses noted in some tick-borne disease patients.

Lyme disease (LD) has been characterized as a multisystem disease caused by the spirochete Borrelia burgdorferi (3). The Centers for Disease Control and Prevention reported 17,730 domestic cases of LD in 2000, making it the most common vector-borne disease in the United States (5). In most patients, antibiotic treatment with doxycycline or amoxicillin has been proven to be a highly effective mode of treatment for acute and late stages of LD (28). However, a subset of patients exhibits persistent symptoms regardless of antibiotic therapeutic intervention.

The primary vertebrate reservoir for B. burgdorferi in the northeastern United States has been identified as the white-footed mouse, Peromyscus leucopus. Both larval and nymphal stages of Ixodes scapularis ticks mainly feed on P. leucopus and can infect human hosts with B. burgdorferi (1). Other pathogens, including Babesia microti and Anaplasma phagocytophila, have also been identified in I. scapularis ticks, and cotransmission with B. burgdorferi has been documented (17, 18, 19).

The present study examined the prevalence of four pathogens in I. scapularis that could potentially be transmitted to humans. Coinfection with two or more of these organisms may complicate LD prognosis. For example, simultaneous LD and babesiosis correlated with a more severe clinical progression than either condition alone (15). I. scapularis ticks were collected by the tick sweep method (4) from February through July 2001, primarily in Union County, N.J., and were identified by standard taxonomic keys (9, 12, 14). Individual ticks were placed in a microcentrifuge tube containing 470 ?l of TE buffer (10 mM Tris-HCl [pH 8.0], 1 mM EDTA), 25 ?l of 10% sodium dodecyl sulfate, and 12 ?l of DNase-free proteinase K (10 mg/ml). A handheld motorized pestle (Kontes, Vineland, N.J.) was used to homogenize each tick lysate. DNA was extracted with phenol-chloroform and recovered by ethanol precipitation. Primers for amplification of genomic DNA were synthesized by Research Genetics (Huntsville, Ala.) and are listed in Table 1. The conditions for PCR amplification of B. burgdorferi, B. microti, Bartonella spp., and A. phagocytophila were as previously described (7, 8, 13, 20). PCR products were analyzed by electrophoresis through a 1% agarose gel containing 0.5-?g/ml ethidium bromide and UV analysis utilizing a Photodocumentation System (Fisher Biotech, Pittsburgh, Pa.). The identity of selected amplicons was confirmed by independent DNA sequencing (SeqWright, Houston, Tex.). Positive controls were included in each PCR experiment and consisted of genomic DNA extracted from pathogens purchased from the American Type Culture Collection (Table 1). Negative controls consisted of the substitution of pyrogen-free water for DNA. Precautions against contamination were maintained as previously described (13).

As another species of tick, Dermacentor variabilis, is also very common in the geographic region studied, molecular techniques were adopted to verify all ticks as I. scapularis (Fig. 1). The primers for amplification were IXO-16S-F and IXO-16S-R (Table 1). The PCR conditions were denaturation at 94°C for 3 min followed by 35 cycles of 94°C for 3 min, 58°C for 1 min, and 72°C for 1 min. Each reaction was concluded with a 10-min final extension step at 72°C in a T3 Thermocycler (Biometra, Göttingen, Germany). Amplification products were resolved through a 2% agarose gel containing 0.5-?g/ml ethidium bromide and visualized on a MultiGenius gel documentation and analysis system (Syngene, Frederick, Md.). The specificity of the primers was verified by amplifying known DNA control extracts from I. scapularis and D. variabilis. Through this technique, 16 ticks were excluded from the sampling pool due to nonamplification. This control also confirms the validity of the results in those samples in which no pathogens were detected. From the PCR analyses, it was evident that at least one of the four pathogens was present in 45.8% (49 of 107) of the ticks (Table 2) Also, 15 of 107 ticks (14.0%) contained more than one pathogen. The most common combination was B. burgdorferi and Bartonella spp.

Transmission of B. burgdorferi to humans causing LD is just one of several possible outcomes from a tick bite. The environment in the tick is suitable for bacterial diversity, and up to 10 clones of B. burgdorferi as well as B. microti and A. phagocytophilum can be simultaneously isolated from a single I. scapularis host (17, 19, 23). In Table 3, the prevalence of the pathogens studied was found to be in agreement with previous reports detailing pathogen detection by PCR methodologies in the northeastern United States, with the exception of the human granulocyltic ehrlichiosis agent, which was at a lower prevalence than all except in northwestern Pennsylvania (11).

This is the first report to assay for Bartonella spp. in field-collected I. scapularis ticks. Previously, we published a clinical case study presenting molecular and serological diagnostic evidence of patients coinfected with B. burgdorferi and Bartonella henselae (13). For two patients, ticks found to contain B. burgdorferi and B. henselae by PCR and identified as I. scapularis were found in their households, one of which was removed from a household cat. Our report was further reinforced by a more recent publication in which both pathogens were detected by PCR in the cerebrospinal fluid of two patients with symptoms suggestive of neuroborreliosis (22). Although the primers in this study were originally selected for the species-specific amplification of B. henselae, this region of the Bartonella 16S ribosomal DNA (rDNA) gene is highly conserved among many species within the genus (2, 21). For example, Multalin alignment analysis revealed that only 5 of 279 nucleotides differ between the B. henselae and B. quintana amplicons, only 2 of which are situated between the oligonucleotide amplification primers (10, 21). Further investigations should seek to amplify more divergent regions of the Bartonella genus that can be utilized for species-specific identification. Members of the Bartonella genus have also been found in ticks and other insects. For example, B. henselae was detected by PCR in 4 of 271 Ixodes ricinus ticks removed from humans in Belluno Province, Italy (24), and Bartonella species were also detected by PCR in 29 of 151 Ixodes pacificus ticks collected in Santa Clara County, Calif. (6).

As treatment for Bartonella infections varies from that prescribed for LD patients, physicians should add Bartonella infections to the list of possible coinfection agents when evaluating patients in regions of tick endemicity, as single- and multiple-pathogen transmission can complicate clinical presentations. Future studies need to clarify that the Bartonella spp. can be passed in culturable form from vector to host and to identify which specific species of Bartonella are present.

Acknowledgments- We thank John F. Anderson (Connecticut Agricultural Experiment Station) for kindly providing the I. scapularis and D. variabilis ticks for molecular identification controls and Chien-Chang Loa and Jason Trama for critical review of the manuscript.


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Figures and Tables

FIG. 1. FIG. 1.Molecular detection of I. scapularis by amplification of 16S rDNA. Lanes: 1, 2, and 5, DNA extracted from I. scapularis; 3 and 4, DNA extracted from D. variabilis; 6, negative water control; M, molecular weight ladder developed in house with bands in (more ...)TABLE 1. TABLE 1. Oligonucleotide primers utilized for species-specific pathogen and l. scapularis amplificationTABLE 2. TABLE 2.Identification of I. scapularis ticks infected with B. burgdorferi, B. microti, A. phagocytophila, or B. henselaeTABLE 3. TABLE 3.

Reported infection rates of B. burgdorferi, A. phagocytophila; and B. microti in the northeastern United States.