Baud, D.

Gerber, S.

Goy, G.

Greub, G.

Hohlfeld, P.

Vial, Y.

Emerging Infectious Diseases

PubMed

Serveur académique lausannois

LETTERSEvidence of Maternal–Fetal Transmission of Parachlamydia acanthamoebaeTo the Editor: Parachlamydia acanthamoebae is a recently identi-fi ed agent of pneumonia (1–3) and has been linked to adverse pregnancy out-comes, including human miscarriage and bovine abortion (4,5). Parachla-mydial sequences have also been de-tected in human cervical smears (4) and in guinea pig inclusion conjunc-tivitis (6). We present direct evidence of maternal–fetal transmission of P. acanthamoebae.We tested 78 amniotic fl uid sam-ples from patients who delivered pre-maturely (defi ned as spontaneous de-livery before 37 weeks of gestation) at the Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland, from 2003 to 2006. DNA was extracted by using the QIAampDNA Mini kit (QIA-GEN, Hilden, Germany) and was test-ed by using a specifi c Parachlamydia real-time PCR (7). One positive sample (threshold cycle of 34.2) was confi rmed by using the 16SigF-Rp2Chlam PCR, which targets a large DNA segment of the 16S rRNA gene (8). Because these 2 PCRs target different DNA segments, the positive PCR results were not due to PCR contamination with amplicons. The sequence exhibited 99.6% similar-ity with P. acanthamoebae strain Hall’s coccus (1) (GenBank accession no. AF366365).The sample was obtained from a 29-year-old woman during her second pregnancy, at 16 weeks of gestation. Amniocentesis was performed because a fi rst-trimester test suggested a Down syndrome risk of 1/100. At the time of amniocentesis, the patient had cough and fl u-like symptoms of 3 weeks’ du-ration, which resolved spontaneously in a few weeks. Cytogenetic analysis showed a normal 46XX karyotype; amniotic fl uid culture remained sterile. The pregnancy ended prematurely at 35 weeks and 6 days with the vaginal delivery of a 2,060-g newborn (<5th percentile). The mother and child had an uneventful hospital course.The role of Parachlamydia as the etiologic agent of premature labor and intrauterine growth retardation is like-ly because 1) all vaginal, placental, and urinary cultures were negative; 2) results of routine serologic tests were negative; and 3) only Parachla-mydia was detected in the amniotic fl uid. Intrauterine infection caused by Parachlamydia spp. may be chronic and asymptomatic until adverse preg-nancy outcomes occur (4).The infection of this pregnant woman might have occurred through zoonotic contact through her work as a butcher in a rural area known for cattle breeding. Of interest, a recent study of 482 healthy Swiss men found 3 who were seropositive for Parachlamydia sp., and all 3 came from the same ru-ral area near Lausanne (within <20-km radius) (9). Moreover, the patient owns 2 guinea pigs, potential vectors of the bacterium (6). Other modes of transmission are possible, e.g., con-taminated water (free-living amebae may serve as hosts for Parachlamydia spp. and are widespread in water net-works) and ingestion of undercooked meat or contaminated cow milk.A plausible pathogenic scenario for this case of possible maternal–fetal transmission of P. acanthamoe-bae might include bacteremia in the context of a lung infection. This could have resulted in intrauterine infection and intrauterine growth restriction.AcknowledgmentsWe warmly thank Gabrielle Gerelle and Philip Tarr for reviewing the manu-script and Sandrine Gremlich-Irrausch and Françoise Damnon for technical help. Research performed at the Center for Research on Intracellular Bacteria is main-ly supported by grants from the Swiss Na-tional Science Foundation (no. FN3200B0-116445), the Faculty of Biology and Medicine of the University of Lausanne (Prix FBM 2006), and Coopération Euro-péenne dans le Domaine de la Recherche Scientifi que et Technique action 855. G.G. is supported by the Leenards Foundation through a career award entitled “Bourse Leenards pour la relève académique en médecine clinique à Lausanne.” This study was approved by the ethics committee of the University of Lausanne.David Baud, Genevieve Goy, Stefan Gerber, Yvan Vial, Patrick Hohlfeld, and Gilbert GreubAuthor affi liations: Institute of Microbiology, Lausanne, Switzerland (D. Baud, G. Goy, G. Greub); and University Hospital of Lau-sanne, Lausanne, Switzerland (D. Baud, S. Gerber, Y. Vial, P. Hohlfeld)DOI: 10.3201/eid1501.080911References  1.  Birtles RJ, Rowbotham TJ, Storey C, Mar-rie TJ, Raoult D. Chlamydia-like obligate parasite of free-living amoebae. Lancet. 1997;349:925–6. DOI: 10.1016/S0140-6736(05)62701-8  2.  G. Greub G, Raoult D. Parachlamydia acanthamoebae, a potential emerging pathogen? Emerg Infect Dis. 2002;8:625–30.  3.  Corsaro D, Greub G. Pathogenic poten-tial of novel Chlamydiae and diagnostic approaches to infections due to these ob-ligate intracellular bacteria. Clin Micro-biol Rev. 2006;19:283–97. DOI: 10.1128/CMR.19.2.283-297.2006  4.  Baud D, Regan L, Greub G. Emerging role of Chlamydia and Chlamydia-like or-ganisms in adverse pregnancy outcomes. Curr Opin Infect Dis. 2008;21:70–6.  5.  Baud D, Thomas V, Arafa A, Regan L, Greub G. Waddlia chondrophila, a po-tential agent of human fetal death. Emerg Infect Dis. 2007;13:1239–43.  6.  Lutz-Wohlgroth L, Becker A, Brugnera E, Huat ZL, Zimmermann D, Grimm F, et al. Chlamydiales in guinea-pigs and their zoonotic potential. J Vet Med A Physiol Pathol Clin Med. 2006;53:185–93. DOI: 10.1111/j.1439-0442.2006.00819.x  7.  Casson N, Posfay-Barbe KM, Gervaix A, Greub G. New diagnostic real-time PCR for specifi c detection of Parachlamydia acanthamoebae DNA in clinical samples. J Clin Microbiol. 2008;46:1491–3. DOI: 10.1128/JCM.02302-07120 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 15, No. 1, January 2009LETTERS  8.  Thomas V, Casson N, Greub G. Cribla-mydia sequanensis, a new intracellular Chlamydiales isolated from Seine river water using amoebal co-culture. Envi-ron Microbiol. 2006;8:2125–35. DOI: 10.1111/j.1462-2920.2006.01094.x  9.  Baud D, Kebbi C, Greub G. Seropreva-lence of different Chlamydia-like organ-isms in an asymptomatic population. Clin Microbiol Infect. In press 2008.Address for correspondence: Gilbert Greub, Center for Research on Intracellular Bacteria, Institute of Microbiology, University Hospital Center and University of Lausanne, Bugnon 48, 1011 Lausanne, Switzerland; email: gilbert.greub@chuv.chEmerging Mycobacteria spp. in Cooling TowersTo the Editor: The importance of nontuberculous mycobacteria (NTM) in various clinical situations recently has increased. Members of the My-cobacterium avium complex (MAC) cause a high percentage of infections in persons with acquired immunode-fi ciency syndrome. Some species are considered emerging pathogens, par-ticularly those of the M. chelonae–M. abscessus group. They occur not only in immunocompromised persons but also in persons without predispos-ing conditions. Although sources of infection are considered to originate from the human environment, until now cooling towers were not clearly demonstrated to be one of these con-tamination sources, despite being sus-pected (1). Natural streams, ground-water, brook waters, and swamps already were reported to contain dif-ferent species of NTM. Constructed environments, such as hospital hot water systems, aerosols from showers, ice machines, swimming pools, dental unit water, endoscopes, and broncho-scopes are other sources (2). Other studies have shown members of MAC in drinking-water distribution systems (3).A 1999 study in South Africa re-ported the presence of mycobacteria in cooling towers but reported no details about species (4). In 2003, cooling towers were reported to be a potential source of slow-growing mycobacterial species (1). Some mycobacterial spe-cies recently were shown to survive within amebae (5). We aimed to study the population of ameba-associated mycobacterial species in 3 cooling towers using a co-coculture method.These cooling towers (E, H, and O), located in downtown Paris, France, were sampled in May 2006. Water was taken in a sample point of the cooling circuit located just before the entrance of the tower basins. Because these cooling towers are regularly treated by oxidizing biocide BCDMH (1-bromo-3-chloro-5, 5-dimethylhydantoin) to prevent development of Legionella spp., no Legionella spp. or Legionella-like amebal pathogens were isolated.Two liters of water samples were fi ltrated through 0.22-μm pore-sized fi lters that were injected onto amebal microplates as previously described (6) and incubated at 32°C. We screened amebal microplates by examination un-der inverted microscope and by Ziehl-Nielsen, Gram, and Gimenez staining. Positive wells were subcultured on ax-enic medium and incubated at 32°C for 10 days. Acid-fast isolates were identi-fi ed by using partial rpoB gene amplifi -cation and sequencing with Myco-F and Myco-R primers (7). These primers did not allow amplifi cation of partial rpoB gene for the 6 M. phocaicum strains. We thus used Myco-F/Myco-Rbon (5′-AGCGGCTGCTGGGTGATCAT- 3′) primer pair for M. phocaicum (8). We compared these sequences with the rpoB gene sequence of Mycobac-teria type strains available in the Gen-eBank database by using BLASTn on the NCBI website (www.ncbi.nlm.nih.gov).We observed bacteria growing in amebae. Subculture on axenic media led to polymicrobial cultures and al-lowed isolation of 33 mycobacterial strains (Table). All these strains were submitted to molecular identifi cation. The 33 isolates corresponded to 5 mycobacterial species: M. fortuitum, M. conceptionense, M. chelonae, M. chimaera, and M. phocaicum (Table). Some of these mycobacteria already had been shown to survive in free-living amebae and to be implicated in human diseases, such as M. chelonae (5). The same author demonstrated recently that 26 environmental myco-bacteria survived in the trophozoites and cysts of Acanthamoeba polypha-ga (5). The recently described spe-cies M. phocaicum was isolated only in samples from humans and was as-sociated with chronic pneumonia (9). However, the natural source of this species is still unknown. M. fortuitum was described as an opportunistic My-cobacterium associated with dissemi-nated infections in a leukemia patient or in furunculosis after footbath in nail  Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 15, No. 1, January 2009 121 Table. Identification of isolated nontuberculous mycobacteria according to their rpoB gene sequence similarity and GenBank accession numbers Closest officially described species (rpoB) Isolates (no.), N = 33 Range in % gene similarity to type strain (GenBank accession no.) Accession numbers ofsequences of isolates Mycobacterium chelonae O (2), E (2) 99.7 (AY147163) EU770577M. conceptionense O (12) 99.2–99.9 (AY859695) EU770583, EU770584 M. fortuitum E (2), H (5), O (3) 99.5–99.7 (AY147165) EU770578, EU770579, EU770580 M. chimaera H (1) 99.7 (EF521908) EU770576M. phocaicum E (5), O (1) 98.3–98.6 (AY859693) EU770581, EU770582 

Evidence of maternal-fetal transmission of Parachlamydia acanthamoebae.

David Baud

Genevieve Goy

Stefan Gerber

Yvan Vial

Patrick Hohlfeld

Gilbert Greub

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Evidence of Maternal–Fetal Transmission of Parachlamydia acanthamoebae

10.1128/JCM.02302-07 120 Emerging Infectious Diseases •

Chlamydia-like obligate parasite of free-living amoebae.

Chlamydiales in guinea-pigs and their zoonotic potential.

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Pathogenic potential of novel Chlamydiae and diagnostic approaches to infections due to these obligate intracellular bacteria. Clin Microbiol Rev.

Waddlia chondrophila, a potential agent of human fetal death. Emerg Infect Dis.

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Evidence of maternal-fetal transmission of Parachlamydia acanthamoebae.

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