Hemoplasma aminations were normal. Abnormal lymph nodes were 2 Infection in cm in diameter, fi rm, and not tender. The patient owned 5
cats and showed signs of multiple cat scratches and bites
HIV-positive on his hands and arms. He had received an HIV-positive Patient, Brazil At admission, his CD4 cell count was 286 cells/mm3
(reference range 500–1,500 cells/mm3), and viral load was 38,100 copies/mL. Additional laboratory fi ndings included
Andrea Pires dos Santos,1
anemia, hematocrit 29% (reference range 38%–50%); leu-
Rodrigo Pires dos Santos,1 Alexander W. Biondo,
kopenia, 3,300 leukocytes/μL (4,300–10,000 leukocytes/
José M. Dora, Luciano Z. Goldani,
μL); thrombocytopenia, 108,000 platelets/μL (150,000–
Simone Tostes de Oliveira,
450,000 platelets/μL); aspartate aminotransferase 66 U/L
Ana Marcia de Sá Guimarães, Jorge Timenetsky,
(15–40 U/L), alanine aminotransferase 79 U/L (10–40
Helio Autran de Morais, Félix H.D. González,
U/L), and lactate dehydrogenase 657 U/L (240–480 U/L).
and Joanne B. Messick
Blood cultures yielded no bacterial growth; sputum cultures were negative for acid-fast bacilli, bacteria, and fungi. Test
Hemotrophic mycoplasmas infect a variety of mam-
results were negative for hepatitis B and C, human T lym-
mals. Although infection in humans is rarely reported, an photropic virus type 1, syphilis, chlamydia, and cryptococ-
association with an immunocompromised state has been cus infections. Bone marrow and lymph node biopsy and
suggested. We report a case of a Mycoplasma haemofelis–
culture results were negative for mycobacterial or fungal
like infection in an HIV-positive patient co-infected with Bar-
Abdominal computed tomography showed hepato-
megaly, splenomegaly, and hypoechoic lesions on the
Hemoplasmas are a group of bacteria that infect animals. spleen. Transesophageal echocardiography showed no le-
They are small epicellular parasites that adhere to the sions compatible with infective endocarditis. An inguinal
host’s erythrocytes. Diseases caused by these bacteria range node biopsy showed granuloma with necrosis suggesting
from acute hemolytic anemia to asymptomatic infection. It cat-scratch disease, and no signs of acid-fast bacilli or
is generally thought that most Mycoplasma spp. are host fungi. Many bacilli suggestive of Bartonella spp. were ob-
specifi c. However, there are occasional reports of infection served by Warthin-Starry staining, and antibodies against
in an animal species not perceived as primary hosts. These B. henselae (titer 256) were detected in serum. Treatment
infections may have a pathologic effect, particularly when with doxycycline was initiated and symptoms subsequently
predisposing conditions, such as immunodefi ciency, are subsided. The patient was discharged and instructed to con-
present (1). We report a case of Mycoplasma haemofelis–
tinue antiretroviral therapy and oral doxycycline.
like infection in an HIV-positive patient with disseminated
In June 2007, the patient was hospitalized with fever,
Bartonella henselae infection.
malaise, weight loss, and lymphadenomegaly. Echocar-diography showed mitral vegetations. Multiple hepatic
The Study
hypoechoic lesions were found by abdominal computed
A 34-year-old HIV-positive man was hospitalized in tomography. Lymph node biopsy specimens showed tiny
Brazil in September 2006 with a 30-day history of night bacilli by Warthin-Starry staining. The patient had prema-
sweats; loss of appetite; productive cough; muscle pain; turely discontinued antiretroviral and antimicrobial drug
and cervical, axillary, and inguinal lymphadenomegaly. treatment, which may have predisposed him to endocarditis
Results of pulmonary, cardiovascular, and abdominal ex-
and hepatic peliosis. He was treated with doxycycline and gentamicin for Bartonella spp. infection. His symptoms
Author affi liations: Universidade Federal do Rio Grande do Sul,
disappeared and an echocardiogram 17 days later showed
Porto Alegre, Rio Grande do Sul, Brazil (A.P. Santos, R.P. Santos,
resolution of mitral vegetations. He was discharged and
J.M. Dora, L. Z. Goldani, S.T. Oliveira, F.H.D. González); Univer-
instructed to continue antiretroviral therapy and oral doxy-
sidade Federal do Paraná, Curitiba, Paraná, Brazil (A.W. Biondo);
cycline for Bartonella spp. infection. Ten months after
University of Illinois, Urbana, Illinois, USA (A.W. Biondo); Univer-
discharge, the patient returned for a follow-up visit while
sidade de São Paulo, São Paulo, Brazil (A.M. de Sá Guimarães,
taking recommended therapy. He had no clinical signs and
J. Timenetsky); University of Wisconsin, Madison, Wisconsin, USA
his laboratory fi ndings were improved.
(H.A. de Morais); and Purdue University, West Lafayette, Indiana,
During the patient’s fi rst hospitalization in 2006, blood
was collected into tubes containing EDTA and 2 aliquots of
1These authors contributed equally to this article.
Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 14, No. 12, December 2008
Hemoplasma Infection in HIV-positive Patient
DNA were extracted (DNeasy Blood and Tissue Kit; QIA-GEN, Valencia, CA, USA) at the Hospital de Clínicas de
Porto Alegre. DNA was tested by PCR for Bartonella spp. infection (2) and by additional PCR protocols for feline he-moplasmas, including Mycoplasma haemofelis (3), “Can-didatus M. haemominutum” (4), and “Candidatus M. turi-censis” (A.P. Santos, unpub. data). These bacteria infect cats and possible infection of this patient was investigated.
Positive controls for M. haemofelis included DNA extract-ed from naturally (GenBank accession no. EU930823) and experimentally infected cats (3). Three positive controls
for B. henselae (type 1, type 2, and Houston strain) were
used. Negative controls included ultrapure water and DNA extracted from blood of a healthy person and a noninfected cat. All negative controls were negative by PCR.
Amplicons of the expected size were obtained in Barto-nella spp. and M. haemofelis PCRs. The 393-bp PCR prod-uct for M. haemofelis (Figure) was purifi ed (Zymoclean Gel DNA Recovery Kit; Zymo Research, Orange, CA, USA), cloned (pGEM-T EasyVector; (Promega, Madison, WI, USA), and sequenced (Purdue Genomics Core Facil-
Figure. PCR results for detection of a Mycoplasma haemofelis–like
ity, West Lafayette, IN, USA). The fragment was 99% ho-
organism in an HIV-positive patient. Lane 1, 100-bp marker; lane
2, positive control (DNA from blood of an M. haemofelis–positive
M. haemofelis 16S rDNA gene sequences in
cat); lane 3, negative control (water); lane 4, DNA from blood of
the GenBank database. To assess the sequence of the 16S the patient.
rRNA gene, we designed species-specifi c primers based on the M. haemofelis sequence (forward primer 5′-ATG CAA GTC GAA CGG ATC TT-3′; reverse primer 5′-TCC AAT associations with latent mycoplasma infections in immu-CAG AAT GTT CAC TC-3′). PCR product amplifi ed from nocompromised and nonimmunocompromised patients are the patient’s blood was purifi ed and sequenced. A 1,214-
now emerging. Increasing numbers of human patients with
bp sequence was submitted to GenBank (accession no. compromised immune systems living near cats increases EU888930); it was 99% homologous with the sequence for the possibility that hemoplasma infections may also emerge M. haemofelis.
The possibility that the patient’s cats might be involved
There are no molecular studies to date documenting
in zoonotic transmission was also investigated. Two weeks hemoplasma infection in humans. However, it has been after the patient’s fi rst blood collection, blood was collected suggested that such infections may be seen in immunocom-from the 5 cats, and DNA was extracted at the Veterinary promised patients (5). A hemotrophic mycoplasma infec-Hospital of the Universidade Federal do Rio Grande do Sul, tion was reported in patients with systemic lupus erythe-Porto Alegre. Two of the cats were positive by PCR for M. matosus (SLE) (6). A 417-bp sequence detected in 1 SLE haemofelis and all 5 cats were positive for Bartonella spp.
patient also showed 99% homology with M. haemofelis (7).
The cats were not tested for other infectious agents.
Sequence data from another hemotrophic mycoplasma in-
PCRs for hemoplasmas were performed in duplicate fection in an anemic human patient were reported in Gen-
at 2 laboratories (Universidade de São Paulo, São Paulo, Bank. However, the sequence of 178 bp of the 16S rRNA Brazil and Purdue University, West Lafayette, IN, USA) by gene (accession no. EU014880) was more closely related using split aliquots. PCR results were reproducible. During to M. suis and M. wenyonii (96%–100%) and only 75% the patient’s second hospitalization, the same PCRs were homologous to M. haemofelis (8). used and the patient was positive for Bartonella spp. but
Epidemiologic studies have linked cat bites and scratch-
es and fl ea-infested cats with transmission of B. henselae to humans (9). B. henselae and B. quintana are causative
Conclusions
agents of bacillary angiomatosis, bacillary peliosis, and
Hemoplasma infections may occur more frequently cat-scratch disease in humans. Peliosis hepatis and lymph
than is generally recognized, given that these organisms node angiomatosis, as seen in this patient, have been asso-fail to grow in culture and only a few laboratories are ciated with B. henselae infection (10). M. haemofelis DNA equipped to detect and identify hemoplasmas (1). Disease has also been detected in cat fl eas (Ctenocephalides felis);
Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 14, No. 12, December 2008
C. felis may be involved in transmission of M. haemofe-
3. Messick JB, Berent LM, Cooper SK. Development and evalua-
lis among cats (11). Additional studies have documented
tion of a PCR-based assay for detection of Haemobartonella felis
experimental transmission by administration of infected
in cats and differentiation of H. felis from related bacteria by re-striction fragment length polymorphism analysis. J Clin Microbiol.
blood intravenously, intraperitoneally, and orally. Hemo-
plasma DNA is present in saliva and feces of cats, which 4. Foley JE, Harrus S, Poland A, Chomel B, Pedersen NC. Molecular, suggests that aggressive interactions among cats involving
clinical and pathologic comparison of two distinct strains of Haemo-
biting may lead to transmission of the organism (
bartonella felis in domestic cats. Am J Vet Res. 1998;59:1581–8. 12). To 5. Duarte MI, Oliveira MS, Shikanai-Yasuda MA, Mariani ON,
our knowledge, there is only 1 other report that cats can be
Takakura CF, Pagliari C, et al. Haemobartonella-like microorganism
co-infected with M. haemofelis and B. henselae (13).
infection in AIDS patients: ultrastructural pathology. J Infect Dis.
As with other Mycoplasma spp., hemoplasmasmight
act as a cofactor in HIV infection, contributing to accelera-
6. Kallick CA, Levin S, Reddi KT, Landau WL. Systemic lupus erythe-
matosus associated with Haemobartonella-like organisms. Nat New
tion of the course of the disease (14). Further studies are
needed to establish the role and prevalence of hemoplasma 7. Kallick CA. Diagnosis of systemic lupus erythematosus, 2005 infection in AIDS patients, as well as the zoonotic potential
[cited 2008 Aug 29]. Available from http://www.wipo.int/pctdb/en/
8. Yang Z, Yan C, Yu F, Hua X. Haemotrophic mycoplasma: review of
aetiology and prevalence of microbial infection. Reviews in Medical
Acknowledgments
We thank the Department of Comparative Pathobiology, 9. Greub G, Raoult D. Bartonella: new explanations for old diseases. J
School of Veterinary Medicine, Purdue University, for laboratory 10. Gasquet S, Maurin M, Brouqui P, Lepidi H, Raoult D. Bacillary an-support and Lynn Guptill for providing positive controls for B.
giomatosis in immunocompromised patients. AIDS. 1998;12:1793–
803. DOI: 10.1097/00002030-199814000-00011
11. Woods JE, Brewer MM, Hawley JR, Wisnewski N, Lappin MR.
A.P.S. was supported by a research fellowship from the Co-
Evaluation of experimental transmission of CandidatusMycoplas-
ordenação de Aperfeiçoamento de Pessoal de Nível Superior. ma haemominutum and Mycoplasma haemofelis by Ctenocepha-lides felis to cats. Am J Vet Res. 2005;66:1008–12. DOI: 10.2460/
Dr A.P. Santos is a doctoral candidate at the Veterinary
School at Universidade Federal do Rio Grande do Sul and a visit-
12. Willi B, Boretti FS, Meli ML, Bernasconi MV, Casati S, Hegglin D,
et al. Real-time PCR investigation of potential vectors, reservoirs,
ing scholar at Purdue University. Her research interests include in-
and shedding patterns of feline hemotropic mycoplasmas. Appl En-
fectious diseases and hemoparasites, particularly hemoplasmas.
viron Microbiol. 2007;73:3798–802. DOI: 10.1128/AEM.02977-06
13. Eberhardt JM, Neal K, Shackelford T, Lappin MR. Prevalence of
selected infectious disease agents in cats from Arizona. J Feline Med
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Address for correspondence: Andrea Pires dos Santos, Department of
uncultured pathogens. N Engl J Med. 1990;323:1573–80.
Veterinary Pathobiology, Purdue University, 625 Harrison St, West Lafayette, IN 47907, USA; email: [email protected]
Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 14, No. 12, December 2008
EFTBA UPDATE – June 2012 EFTBA Activity The EFTBA AGM on the 13th May was well attended in the magnificent location of Longchamp racecourse in Paris. Many issues of current concern and relevance were debated and the packed AGM agenda was followed by racing in the afternoon including a prize giving to Dietrich von Boetticher from Germany as the 2012 EFTBA award winner. Following the AGM m
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