Bartonella species in blood of immunocompetent persons with animal and arthropod contact.Using PCR PCR polymerase chain reaction.
polymerase chain reaction
Polymerase chain reaction (PCR) in conjunction with pre-enrichment culture, we detected Bartonella henselae Bartonella henselae Rochalimaea henselae Infectious disease A slender, fastidious coccobacillary bacterium of the normal flora of cats associated with bacteremia, endocarditis, cat-scratch disease, bacillary angiomatosis, peliosis hepatis; it may affect and B. vinsonii subspecies subspecies, also called race, a genetically distinct geographical subunit of a species. See also classification. berkhoffii in the blood of 14 immunocompetent im·mu·no·com·pe·tent
Having the normal bodily capacity to develop an immune response following exposure to an antigen.
im persons who had frequent animal contact and arthropod arthropod
Any member of the largest phylum, Arthropoda, in the animal kingdom. Arthropoda consists of more than one million known invertebrate species in four subphyla: Uniramia (five classes, including insects), Chelicerata (three classes, including arachnids and horseshoe exposure.
Attempts to isolate Bartonella sp. from immunocompetent persons with serologic se·rol·o·gy
n. pl. se·rol·o·gies
1. The science that deals with the properties and reactions of serums, especially blood serum.
2. , pathologic, or molecular evidence of infection are often unsuccessful; several investigators have indicated that Bartonella isolation methods need to be improved (1-4). By combining PCR and pre-enrichment culture, we detected B. henselae and B. vinsonii subspecies berkhoffii infection in the blood of immunocompetent persons who had arthropod and occupational animal exposure.
From November 2004 through June 2005, blood and serum samples from 42 persons were tested, and 14 completed a questionnaire, approved by the North Carolina State University History
A nutrient culture medium that is enriched with whole blood and used for the growth of certain strains of bacteria. plate (Figure). An uninoculated, pre-enrichment culture was processed simultaneously as a control. Methods used for DNA extraction and conventional and real-time PCR targeting of the Bartonella 16S-23S intergenic spacer (ITS) region and heme-binding protein (Pap31) gene have been described (7,8). Conventional PCR amplicons were cloned with the pGEM-T Easy Vector System (Promega, Madison, WI, USA); sequencing was performed by Davis Sequencing, Inc. (Davis, CA, USA). Sequences were aligned and compared with GenBank sequences with AlignX software (Vector NTI NTI NewTech Infosystems (software company, Irvine, California)
NTI Nuclear Threat Initiative
NTI National Transit Institute (New Brunswick, New Jersey)
NTI Nunavut Tunngavik Incorporated Suite 6.0 (InforMax, Inc., Bethesda, MD, USA) (7,8). B. vinsonii subsp. berkhoffii, B. henselae, and B. quintana antibodies were determined by using a modification of a previously described immunofluorescence Immunofluorescence
A technique that uses a fluorochrome to indicate the occurrence of a specific antigen-antibody reaction. The fluorochrome labels either an antigen or an antibody. antibody assay (IFA Immunofluorescent assay (IFA)
A blood test sometimes used to confirm ELISA results instead of using the Western blotting. In an IFA test, HIV antigen is mixed with a fluorescent compound and then with a sample of the patient's blood. ) procedure (9).
Study participants included 12 women and 2 men, ranging in age from 30 to 53 years; all of them reported occupational animal contact for >10 years (Table). Most had daily contact with cats (13 persons) and dogs (12 persons). All participants reported animal bites or scratches (primarily from cats) and arthropod exposure, including fleas, ticks, biting flies, mosquitoes, lice, mites, or chiggers chiggers Harvest mites, red mites Dermatology Larvae of the family Trombiculidae, genus Eutrombicula–southern US, Trombicula–Europe which causes skin infestation Habitat Berry patches, tall grass, weeds, woods. Cf Chiggers. . All participants reported intermittent or chronic clinical symptoms, including fatigue, arthralgia arthralgia /ar·thral·gia/ (ahr-thral´jah) pain in a joint.
Severe pain in a joint. Also called arthrodynia. , myalgia myalgia /my·al·gia/ (mi-al´jah) muscular pain.myal´gic
epidemic myalgia see under pleurodynia.
n. , headache, memory loss, ataxia ataxia (ətăk`sēə), lack of coordination of the voluntary muscles resulting in irregular movements of the body. Ataxia can be brought on by an injury, infection, or degenerative disease of the central nervous system, e.g. , and paresthesia paresthesia /par·es·the·sia/ (par?es-the´zhah) morbid or perverted sensation; an abnormal sensation, as burning, prickling, formication, etc.
par·es·the·sia or par·aes·the·sia
n. (Table). Illness was most frequently mild to moderate in severity, with a waxing and waning course, and all but 2 persons could perform occupational activities. Of the 14 participants, 9 had been evaluated by a cardiologist, 8 each by an infectious disease Infectious disease
A pathological condition spread among biological species. Infectious diseases, although varied in their effects, are always associated with viruses, bacteria, fungi, protozoa, multicellular parasites and aberrant proteins known as prions. physician or a neurologist, and 5 each by an internist or a rheumatologist rheumatologist /rheu·ma·tol·o·gist/ (roo?mah-tol´ah-jist) a specialist in rheumatology.
A specialist in the diagnosis and treatment of rheumatic disorders. . Eleven participants had received antimicrobial drugs.
When reciprocal titers of [greater than or equal to] 64 were used, 8 persons were seroreactive to Bartonella antigens (online Appendix Table, available from www.cdc.gov/eid/content/13/6/938appT.htm). B. henselae or B. vinsonii subsp, berkhoffii was detected or isolated from all 14 participants. At the time of initial testing, Bartonella DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. was amplified directly from 3 blood samples, from 7 pre-enrichment liquid cultures, and from 4 subculture isolates (Online Table). For 5 persons, results of PCR and culture of initial samples were negative. Overall, Bartonella DNA was amplified from 11 (28%) of 40 extracted blood samples, 13 (33%) of 40 pre-enrichment cultures, and 5 isolates. For 7 persons, B. henselae DNA was amplified at multiple time points. Bartonella DNA was never amplified from any PCR control or uninoculated culture control.
By using the ITS target region, 2 distinct B. henselae ITS and Pap31 strains were sequenced, B. henselae Houston I (HI) (GenBank NC-005956) and B. henselae San Antonio 2 (SA2) (GenBank AF369529). Within the noncoding ITS region, B. henselae SA2 strains have a 30-bp insertion (ATT ATT
ammonia tolerance test. GCT (programming, tool) GCT - A test-coverage tool by Brian Marick <firstname.lastname@example.org>, based on GNU C. Version 1.4 was ported to Sun-3, Sun-4, RS/6000, 68000, 88000, HP-PA, IBM 3090, Ultrix, Convex, SCO but not Linux, Solaris, or Microsoft Windows. TCT TCT The Capital Times (Madison, WI newspaper)
TCT Transcatheter Cardiovascular Therapeutics
TCT The Coroner's Toolkit
TCT Trans Canada Trail
TCT Tcl Core Team
TCT Tsukuba College of Technology (Japan) AAA AAA: see American Automobile Association.
(Triple A) A common single-cell battery used in a myriad of electronic devices of all variety. Like its double A (AA) cousin, it provides 1.5 volts of DC power. When used in series, the voltage is multiplied. AAG AAG Association of American Geographers (Washington, DC)
AAG Assistant Attorney General
AAG Asociación Argentina de Golf
AAG Anti-Aircraft Gun
AAG Assistant Adjutant General
AAG Australian Association of Gerontology ATT GCT TCT AAA AAG) located 518 bases downstream from the 16S gene. Only B. vinsonii subsp, berkhoffii types I and II were detected (8).
Persistent human infection with B. bacilliformis and B. quintana has been previously documented, whereas infection with B. henselae (cat-scratch disease Cat-Scratch Disease Definition
Cat-scratch disease is an uncommon infection that typically results from a cat's scratch or bite. Most sufferers experience only moderate discomfort and find that their symptoms clear up without any lasting harm after a [CSD CSD Commission on Sustainable Development
CSD Serbian Dinar (ISO currency code)
CSD Christopher Street Day
CSD Circuit Switched Data (Sprint)
CSD Computer Science Department
CSD Community School District ]) is generally considered self-limiting (1,2,10). Recently, B. henselae DNA was amplified from the blood of a child 4 months after CSD diagnosis (11). Our study indicates that B. henselae and B. vinsonii subsp, berkhoffii can induce occult infection occult infection An infection first recognized by 2º manifestations–eg, ↑ PMNs in the circulation or FUO, often caused by a bacterial infection in an obscure site–eg, a subphrenic or other intraabdominal region. Cf Fever of unknown origin. in immunocompetent persons and that detection can be enhanced by combining PCR with pre-enrichment culture. Considering only the results from initial blood samples, PCR detected Bartonella DNA in 3 samples, all of which were subsequently PCR positive by subculture or enrichment culture. In samples from 5 persons, pre-enrichment was necessary, and in 5 other persons, sequential sampling was necessary to detect Bartonella infection. Intermittent bacteremia Intermittent bacteremia
A kind of bacteremia where the bacteria enter the blood at various time intervals.
Mentioned in: Blood Culture , as occurs in B. henselae-infected cats (12), antimicrobial drug administration, low bacterial copy numbers, and low inoculum inoculum /in·oc·u·lum/ (-ok´u-lum) pl. inoc´ula material used in inoculation.
n. pl. volume (1 mL) may have contributed to intermittent detection or inability to isolate Bartonella spp. from some participant samples. Although our approach is an improvement over historical isolation approaches, our results emphasize ongoing limitations associated with the detection of Bartonella infection. Obtaining stable Bartonella subcultures (n = 5 in this study) has proven problematic for other specialized laboratories that routinely culture for Bartonella spp. (3,4). To our knowledge, the B. vinsonii subsp, berkhoffii type II isolate described in our study is the only type II human isolate reported to date (8). Various combinations of B. henselae and B. vinsonii subsp, berkhoffii strain types were detected in the same blood sample or sequential blood samples. The coexistence of B. henselae genetic variants has been described among primary patient isolates, which suggests that multiple genotypes may emerge within the same person (13).
Overall, 57% of persons tested were seroreactive to 1 or all 3 Bartonella test antigens. Previous reports from the United States identified a B. henselae seroprevalence seroprevalence Immunology The proportion of a population that is seropositive–ie, has been exposed to a particular pathogen or immunogen; the seropositivity of a population is calculated as the number of individuals who produce a particular antibody divided of 3% in healthy blood donors and a cumulative seroprevalence of 7.1% to both B. henselae and B. quintana antigens in veterinary professionals (1). In this and other studies, serologic test serologic test Lab medicine A test that measures components–eg, antibodies, complement, and reactions–eg, complement fixation, agglutination, precipitation, etc, that reflect immune status, especially antibody titers. Cf Seroconversion. results did not correlate with PCR amplification or isolation results. Antigenic variability among B. henselae test strains can cause false-negative IFA results in persons with suspected CSD. Also B. henselae, B. quintana, or B. elizabethae antibodies were not detected in some persons with DNA evidence Among the many new tools that science has provided for the analysis of forensic evidence is the powerful and controversial analysis of deoxyribonucleic acid, or DNA, the material that makes up the genetic code of most organisms. of active infection (1,3,4).
Animal contact, often to a wide spectrum of domestic and wild animal species, is an obvious consequence of the daily activities of the study population, which is biased by veterinary occupational exposure and by self-selection (volunteer bias). Cats are considered the primary reservoir host reservoir host
A host that serves as a source of infection and potential reinfection of humans and as a means of sustaining a parasite when it is not infecting humans. for B. henselae, whereas coyotes and foxes are considered reservoir hosts for B. vinsonii subsp, berkhoffii (1,2,8). Detection of B. vinsonii subsp, berkhoffii in 4 of 5 Californian participants could be related to the high prevalence of bacteremic bac·te·re·mi·a
The presence of bacteria in the blood.
bacte·re coyotes in this region as well as to the potential transmission by a tick vector (1,2). All 14 participants reported frequent arthropod exposure. Although Bartonella spp.transmission by ticks has not been proven, several recent studies have identified Bartonella DNA in questing ticks, ticks attached to animals, and ticks attached to humans (1,2,14).
Despite reporting chronic or episodic illness, most participants continued to effectively maintain daily professional and personal activities. The symptoms described in the study patients are very similar to those described in a community and hospital-based surveillance study of CSD patients, in whom CSD-associated arthropathy arthropathy /ar·throp·a·thy/ (ahr-throp´ah-the) any joint disease.arthropath´ic
Charcot's arthropathy neuropathic a. was an uncommon chronic syndrome affecting mostly young and middle-age women (15). Our study was initiated to investigate the feasibility of combining PCR with pre-enrichment culture. Prospective studies, with appropriate controls, are needed to characterize the prevalence and clinical relevance of persistent Bartonella infection in immunocompetent persons.
We thank the study participants for providing blood samples, Julie Bradley and Maria Belen Cadenas for technical assistance, and Tonya Lee for editorial assistance.
This research was supported by the state of North Carolina North Carolina, state in the SE United States. It is bordered by the Atlantic Ocean (E), South Carolina and Georgia (S), Tennessee (W), and Virginia (N). Facts and Figures
Area, 52,586 sq mi (136,198 sq km). Pop. and, in part, through a gift from Bayer Animal Health (to R.G.M. and A.W.D.).
(1.) Chomel BB, Kasten RW, Sykes JE, Boulouis H J, Breitschwerdt EB. Clinical impact of persistent Bartonella bacteremia bacteremia: see septicemia.
Presence of bacteria in the blood. Short-term bacteremia follows dental or surgical procedures, especially if local infection or very high-risk surgery releases bacteria from isolated sites. in humans and animals. Ann NY Acad Sci. 2003;990:267-78.
(2.) Boulouis H-J, Chang CC, Henn JB, Kasten RW, Chomel BB. Factors associated with the rapid emergence of zoonotic Zoonotic
A disease which can be spread from animals to humans.
Mentioned in: Zoonosis Bartonella infections. Vet Res. 2005;36:383-410.
(3.) La Scola B, Raoult D. Culture of Bartonella quintana and Bartonella henselae from human samples: a 5-year experience (1993-1998). J Clin Microbiol. 1999;37:1899-905.
(4.) Gouriet F, Fenollar F, Patrice JY, Dancourt M, Raoult D. Use of shell-vial cell culture assay for isolation of bacteria from clinical specimens: 13 years of experience. J Clin Microbiol. 2005;43:4993-5002.
(5.) Maggi RG, Harms CA, Hohn AA, Pabst DA, McLellan WA, Walton WJ, et al. Bartonella henselae in porpoise porpoise, small whale of the family Phocaenidae, allied to the dolphin. Porpoises, like other whales, are mammals; they are warm-blooded, breathe air, and give birth to live young, which they suckle with milk. blood. Emerg Infect Dis. 2005;11:1894-8.
(6.) Breitschwerdt EB, Maggi RG, Sigmon B, Nicholson WL. Isolation of Bartonella quintana from a woman and a cat following putative bite transmission. J Clin Microbiol. 2007;45:270-2.
(7.) Maggi RG, Breitschwerdt EB. Potential limitations of the 16S-23S rRNA intergenic region for the molecular detection of Bartonella species. J Clin Microbiol. 2005;43:1171-6.
(8.) Maggi RG, Chomel B, Hegarty BC, Henn J, Breitschwerdt EB. A Bartonella vinsonii berkhoffii typing scheme based upon 16S-23S ITS and Pap31 sequences from dog, coyote coyote (kī`ōt, kīō`tē) or prairie wolf, small, swift wolf, Canis latrans, native to W North America. It is found in deserts, prairies, open woodlands, and brush country; it is also called brush wolf. , gray fox, and human isolates. Mol Cell Probes. 2006;20:128-34.
(9.) Dalton MJ, Robinson LE, Copper J, Regnery RL, Olson JG, Childs JE. Use of Bartonella antigens for serologic diagnosis of cat-scratch disease at a national referral center. Arch Intern Med. 1995; 155:1670-6.
(10.) Brouqui P, La Scola B, Roux Roux , Pierre Paul Émile 1853-1933.
French bacteriologist. His work with the diphtheria bacillus led to the development of antitoxins to neutralize pathogenic toxins. V, Raoult D. Chronic Bartonella quintana bacteremia in homeless patients. N Engl J Med. 1999;340: 184-9.
(11.) Arvand M, Schad SG. Isolation of Bartonella henselae DNA from the peripheral blood of a patient with cat scratch disease cat scratch disease
An infectious disease that may follow the scratch or bite of a cat, producing localized inflammation of lymph nodes and a low-grade fever. Also called benign inoculation lymphoreticulosis, cat scratch fever. up to 4 months after the cat scratch injury. J Clin Microbioh 2006;44: 2288-90.
(12.) Kordick DL, Brown TT, Shin KO, Breitschwerdt EB. Clinical and pathological evaluation of chronic Bartonella henselae or Bartonella elarridgeiae infection in cats. J Clin Microbiol. 1999;37:1536-47.
(13.) Arvand M, Schubert H, Viezens J. Emergence of distinct genetic variants in the population of primary Bartonella henselae isolates. Microbes Infect. 2006;8:1315-20.
(14.) Adelson ME, Rao RV, Tilton RC, Cabets K, Eskow E, Fein L, et al. Prevalence of Borrelia burgdorferi, Bartonella spp., Babesia microti, and Anaplasma phagocytophila in Ixodes scapularis ticks collected in Northern New Jersey. J Clin Microbiol. 2004;42:2799-80l.
(15.) Giladi M, Maman E, Paran D, Bickels J, Comaneshter D, Avidor B, et al. Cat-scratch disease-associated arthropathy. Arthritis Rheum rheum (rldbomacm) any watery or catarrhal discharge.
A watery or thin mucous discharge from the eyes or nose.
any watery or catarrhal discharge. . 2005;52:3611-7.
Edward B. Breitschwerdt, * Ricardo G. Maggi, * Ashlee W. Duncan, * William L. Nicholson, ([dagger]) Barbara C. Hegarty, * and Christopher W. Woods ([double dagger])
* North Carolina State University College of Veterinary Medicine, Raleigh, North Carolina For other uses of this name, see Raleigh.
Raleigh (IPA: /ˈrɑli/, ral-ee) is the capital of the State of North Carolina and the county seat of Wake County. , USA, ([dagger]) Centers for Disease Control and Prevention Centers for Disease Control and Prevention (CDC), agency of the U.S. Public Health Service since 1973, with headquarters in Atlanta; it was established in 1946 as the Communicable Disease Center. , Atlanta, Georgia, USA; and ([double dagger]) Duke University Medical Center, Durham, North Carolina Durham is a city in the U.S. state of North Carolina. It is the county seat of Durham CountyGR6 and is the fourth-largest city in the state by population. , USA
Dr Breitschwerdt is a professor of medicine and infectious diseases at the College of Veterinary Medicine, North Carolina State University. He is also adjunct associate professor of medicine at Duke University Medical Center. His research focuses on comparative medical aspects of zoonotic vectorborne infections in cats, dogs, and humans.
Address for correspondence: Edward B. Breitschwerdt, North Carolina State University College of Veterinary Medicine, 4700 Hillsborough St, Raleigh, NC 27606, USA; email: email@example.com
Table. Selected demographic, epidemiologic, and medical information reported by 14 immunocompetent persons infected with Bartonella henselae or B. vinsonii subsp. berkhoffii * Study participant no. Characteristic/ symptom 1 2 3 4 Sex F F F F Age, y 51 30 48 44 State of NC NC NC CO residence Occupational V VtA AHR V animal exposure Daily contact Y/Y Y/N N/Y Y/Y with dogs/cats Contact with 2/1 3/3 4/4 4/4 fleas/ticks ([dagger]) Self-health CI CI II II assessment ([double dagger]) Fatigue + + - + Joint pain + + - + Difficulty + + - - sleeping (insomnia) Muscle pain + + - - Difficulty + + - - remembering Loss of + + + - sensation or numbness Balance + - - problems Headache + + - - Tremors + - - Irritability + - - Bowel or + - + bladder dysfunction Eye pain + - + Blurred vision + - - Sleepiness + - - Syncope or + + + - fainting episodes Shortness of + - + breath Muscle + - - weakness Study participant no. Characteristic/ symptom 5 6 7 8 Sex F F M F Age, y 53 50 32 33 State of VA CA NC VA residence Occupational V CR VtA VtA animal exposure Daily contact Y/Y Y/Y Y/Y Y/Y with dogs/cats Contact with 3/3 2/3 3/3 2/2 fleas/ticks ([dagger]) Self-health II CI CI CI assessment ([double dagger]) Fatigue + + + + Joint pain + + + Difficulty + - + + sleeping (insomnia) Muscle pain U + + Difficulty + - + + remembering Loss of + - + sensation or numbness Balance + + + + problems Headache + + + Tremors - + + Irritability - + + Bowel or + - + bladder dysfunction Eye pain + + + Blurred vision - + + Sleepiness - + - Syncope or + + fainting episodes Shortness of - + + breath Muscle + weakness Study participant no. Characteristic/ symptom 9 10 11 12 Sex F F F F Age, y 48 53 52 39 State of CA CA CA CA residence Occupational VtA VtA VtA V animal exposure Daily contact Y/Y Y/Y Y/Y Y/Y with dogs/cats Contact with 4/4 2/4 3/1 3/2 fleas/ticks ([dagger]) Self-health CI II II CI assessment ([double dagger]) Fatigue + + + + Joint pain - + + + Difficulty + + + + sleeping (insomnia) Muscle pain - + U + Difficulty + + + - remembering Loss of - + + sensation or numbness Balance - + + problems Headache - + Tremors - + + + Irritability + + - Bowel or - + - bladder dysfunction Eye pain - Blurred vision - + + - Sleepiness - + + - Syncope or fainting episodes Shortness of - + - breath Muscle - + + + weakness Study participant no. Characteristic/ Total, symptom 13 14 N = 14 Sex M F Age, y 52 44 State of VA MN residence Occupational WB WB animal exposure Daily contact N/Y Y/Y with dogs/cats Contact with NA/3 4/3 fleas/ticks ([dagger]) Self-health II CI assessment ([double dagger]) Fatigue + + 13 Joint pain + U 10 Difficulty - - 9 sleeping (insomnia) Muscle pain + + 8 Difficulty - U 8 remembering Loss of - U 7 sensation or numbness Balance - - 7 problems Headache + U 7 Tremors - - 6 Irritability - + 6 Bowel or + - 6 bladder dysfunction Eye pain - 5 Blurred vision - - 5 Sleepiness - + 5 Syncope or 5 fainting episodes Shortness of - U 5 breath Muscle - U 5 weakness * F, female; M, male; NC, North Carolina, CO, Colorado, VA, Virginia; CA, California, MN, Minnesota; V, veterinarian; VtA, veterinary assistant; AHR, animal health researcher; CR, cattle rancher; WB, wildlife biologist; Y, yes; N, no, with respect to the study participant's daily contact with dogs/cats; CI, chronically ill; II, infrequently ill; +, yes; -, no; blank, no answer reported; U, unknown. ([dagger]) Reported as frequencies and defined as follows: 1, daily; 2, infrequently (weekly); 3, occasionally (monthly); 4, almost never (yearly). ([double dagger]) Self-health assessment: As part of the questionnaire, study participants were asked to rate their own health status: healthy, infrequently ill, or chronically ill.