LETTERS

Evidence of The pregnancy ended prematurely at Medicine of the University of Lausanne 35 weeks and 6 days with the vaginal (Prix FBM 2006), and Coopération Euro- Maternal–Fetal delivery of a 2,060-g newborn (<5th péenne dans le Domaine de la Recherche Transmission of percentile). The mother and child had Scientifi que et Technique action 855. G.G. Parachlamydia an uneventful hospital course. is supported by the Leenards Foundation The role of Parachlamydia as the through a career award entitled “Bourse acanthamoebae etiologic agent of premature labor and Leenards pour la relève académique en To the Editor: Parachlamydia intrauterine growth retardation is like- médecine clinique à Lausanne.” This study acanthamoebae is a recently identi- ly because 1) all vaginal, placental, was approved by the ethics committee of fi ed agent of pneumonia (1–3) and has and urinary cultures were negative; the University of Lausanne. been linked to adverse pregnancy out- 2) results of routine serologic tests comes, including human miscarriage were negative; and 3) only Parachla- David Baud, Genevieve Goy, and bovine abortion (4,5). Parachla- mydia was detected in the amniotic Stefan Gerber, Yvan Vial, mydial sequences have also been de- fl uid. Intrauterine infection caused by Patrick Hohlfeld, tected in human cervical smears (4) Parachlamydia spp. may be chronic and Gilbert Greub and in guinea pig inclusion conjunc- and asymptomatic until adverse preg- Author affi liations: Institute of Microbiology, tivitis (6). We present direct evidence nancy outcomes occur (4). Lausanne, Switzerland (D. Baud, G. Goy, of maternal–fetal transmission of P. The infection of this pregnant G. Greub); and University Hospital of Lau- acanthamoebae. woman might have occurred through sanne, Lausanne, Switzerland (D. Baud, We tested 78 amniotic fl uid sam- zoonotic contact through her work as a S. Gerber, Y. Vial, P. Hohlfeld) ples from patients who delivered pre- butcher in a rural area known for cattle DOI: 10.3201/eid1501.080911 maturely (defi ned as spontaneous de- breeding. Of interest, a recent study of livery before 37 weeks of gestation) 482 healthy Swiss men found 3 who References at the Centre Hospitalier Universitaire were seropositive for Parachlamydia sp., and all 3 came from the same ru- Vaudois, Lausanne, Switzerland, from 1. Birtles RJ, Rowbotham TJ, Storey C, Mar- 2003 to 2006. DNA was extracted by ral area near Lausanne (within <20- rie TJ, Raoult D. Chlamydia-like obligate using the QIAampDNA Mini kit (QIA- km radius) (9). Moreover, the patient parasite of free-living amoebae. Lancet. 1997;349:925–6. DOI: 10.1016/S0140- GEN, Hilden, Germany) and was test- owns 2 guinea pigs, potential vectors of the bacterium (6). Other modes of 6736(05)62701-8 ed by using a specifi c Parachlamydia 2. G. Greub G, Raoult D. Parachlamydia real-time PCR (7). One positive sample transmission are possible, e.g., con- acanthamoebae, a potential emerging (threshold cycle of 34.2) was confi rmed taminated water (free-living amebae pathogen? Emerg Infect Dis. 2002;8: 625–30. by using the 16SigF-Rp2Chlam PCR, may serve as hosts for Parachlamydia spp. and are widespread in water net- 3. Corsaro D, Greub G. Pathogenic poten- which targets a large DNA segment of tial of novel and diagnostic the 16S rRNA gene (8). Because these works) and ingestion of undercooked approaches to infections due to these ob- 2 PCRs target different DNA segments, meat or contaminated cow milk. ligate intracellular . Clin Micro- biol Rev. 2006;19:283–97. DOI: 10.1128/ the positive PCR results were not due A plausible pathogenic scenario for this case of possible maternal– CMR.19.2.283-297.2006 to PCR contamination with amplicons. 4. Baud D, Regan L, Greub G. Emerging The sequence exhibited 99.6% similar- fetal transmission of P. acanthamoe- role of and Chlamydia-like or- ity with P. acanthamoebae strain Hall’s bae might include bacteremia in the ganisms in adverse pregnancy outcomes. Curr Opin Infect Dis. 2008;21:70–6. coccus (1) (GenBank accession no. context of a lung infection. This could have resulted in intrauterine infection 5. Baud D, Thomas V, Arafa A, Regan L, AF366365). Greub G. chondrophila, a po- The sample was obtained from a and intrauterine growth restriction. tential agent of human fetal death. Emerg 29-year-old woman during her second Infect Dis. 2007;13:1239–43. Acknowledgments 6. Lutz-Wohlgroth L, Becker A, Brugnera pregnancy, at 16 weeks of gestation. We warmly thank Gabrielle Gerelle E, Huat ZL, Zimmermann D, Grimm F, et Amniocentesis was performed because and Philip Tarr for reviewing the manu- al. Chlamydiales in guinea-pigs and their a fi rst-trimester test suggested a Down script and Sandrine Gremlich-Irrausch and zoonotic potential. J Vet Med A Physiol syndrome risk of 1/100. At the time of Pathol Clin Med. 2006;53:185–93. DOI: Françoise Damnon for technical help. 10.1111/j.1439-0442.2006.00819.x amniocentesis, the patient had cough 7. Casson N, Posfay-Barbe KM, Gervaix A, Research performed at the Center for and fl u-like symptoms of 3 weeks’ du- Greub G. New diagnostic real-time PCR ration, which resolved spontaneously Research on Intracellular Bacteria is main- for specifi c detection of Parachlamydia in a few weeks. Cytogenetic analysis ly supported by grants from the Swiss Na- acanthamoebae DNA in clinical samples. tional Science Foundation (no. FN3200B0- J Clin Microbiol. 2008;46:1491–3. DOI: showed a normal 46XX karyotype; 10.1128/JCM.02302-07 amniotic fl uid culture remained sterile. 116445), the Faculty of Biology and

120 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 15, No. 1, January 2009 LETTERS

8. Thomas V, Casson N, Greub G. Cribla- water, brook waters, and swamps enic medium and incubated at 32°C for mydia sequanensis, a new intracellular already were reported to contain dif- 10 days. Acid-fast isolates were identi- Chlamydiales isolated from Seine river water using amoebal co-culture. Envi- ferent species of NTM. Constructed fi ed by using partial rpoB gene amplifi - ron Microbiol. 2006;8:2125–35. DOI: environments, such as hospital hot cation and sequencing with Myco-F and 10.1111/j.1462-2920.2006.01094.x water systems, aerosols from showers, Myco-R primers (7). These primers did 9. Baud D, Kebbi C, Greub G. Seropreva- ice machines, swimming pools, dental not allow amplifi cation of partial rpoB lence of different Chlamydia-like organ- isms in an asymptomatic population. Clin unit water, endoscopes, and broncho- gene for the 6 M. phocaicum strains. Microbiol Infect. In press 2008. scopes are other sources (2). Other We thus used Myco-F/Myco-Rbon studies have shown members of MAC (5′-AGCGGCTGCTGGGTGATCAT- Address for correspondence: Gilbert Greub, in drinking-water distribution systems 3′) primer pair for M. phocaicum (8). Center for Research on Intracellular Bacteria, (3). We compared these sequences with Institute of Microbiology, University Hospital A 1999 study in South Africa re- the rpoB gene sequence of Mycobac- Center and University of Lausanne, Bugnon ported the presence of mycobacteria in teria type strains available in the Gen- 48, 1011 Lausanne, Switzerland; email: gilbert. cooling towers but reported no details eBank database by using BLASTn on [email protected] about species (4). In 2003, cooling the NCBI website (www.ncbi.nlm.nih. towers were reported to be a potential gov). source of slow-growing mycobacterial We observed bacteria growing in species (1). Some mycobacterial spe- amebae. Subculture on axenic media cies recently were shown to survive led to polymicrobial cultures and al- within amebae (5). We aimed to study lowed isolation of 33 mycobacterial Emerging the population of ameba-associated strains (Table). All these strains were Mycobacteria spp. mycobacterial species in 3 cooling submitted to molecular identifi cation. in Cooling Towers towers using a co-coculture method. The 33 isolates corresponded to 5 These cooling towers (E, H, and mycobacterial species: M. fortuitum, To the Editor: The importance of O), located in downtown Paris, France, M. conceptionense, M. chelonae, M. nontuberculous mycobacteria (NTM) were sampled in May 2006. Water was chimaera, and M. phocaicum (Table). in various clinical situations recently taken in a sample point of the cooling Some of these mycobacteria already has increased. Members of the My- circuit located just before the entrance had been shown to survive in free- cobacterium avium complex (MAC) of the tower basins. Because these living amebae and to be implicated in cause a high percentage of infections cooling towers are regularly treated by human diseases, such as M. chelonae in persons with acquired immunode- oxidizing biocide BCDMH (1-bromo- (5). The same author demonstrated fi ciency syndrome. Some species are 3-chloro-5, 5-dimethylhydantoin) to recently that 26 environmental myco- considered emerging pathogens, par- prevent development of Legionella bacteria survived in the trophozoites ticularly those of the M. chelonae–M. spp., no Legionella spp. or Legionella- and cysts of Acanthamoeba polypha- abscessus group. They occur not only like amebal pathogens were isolated. ga (5). The recently described spe- in immunocompromised persons but Two liters of water samples were cies M. phocaicum was isolated only also in persons without predispos- fi ltrated through 0.22-μm pore-sized in samples from humans and was as- ing conditions. Although sources of fi lters that were injected onto amebal sociated with chronic pneumonia (9). infection are considered to originate microplates as previously described (6) However, the natural source of this from the human environment, until and incubated at 32°C. We screened species is still unknown. M. fortuitum now cooling towers were not clearly amebal microplates by examination un- was described as an opportunistic My- demonstrated to be one of these con- der inverted microscope and by Ziehl- cobacterium associated with dissemi- tamination sources, despite being sus- Nielsen, Gram, and Gimenez staining. nated infections in a leukemia patient pected (1). Natural streams, ground- Positive wells were subcultured on ax- or in furunculosis after footbath in nail

Table. Identification of isolated nontuberculous mycobacteria according to their rpoB gene sequence similarity and GenBank accession numbers Closest officially described Range in % gene similarity to type Accession numbers of species (rpoB) Isolates (no.), N = 33 strain (GenBank accession no.) sequences of isolates Mycobacterium chelonae O (2), E (2) 99.7 (AY147163) EU770577 M. 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) EU770576 M. phocaicum E (5), O (1) 98.3–98.6 (AY859693) EU770581, EU770582

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