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Novel Chlamydiaceae Disease in Captive Salamanders

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Novel Chlamydiaceae Disease in Captive Salamanders LETTERS health concern in industrialized and Author affi liations: Institute of Tropical 8. Ewers C, Grobbel M, Stamm I, Kopp PA, resource-poor settings. Few reports Medicine, Antwerp, Belgium (H. De Boeck, Diehl I, Semmler T, et al. Emergence of human pandemic O25:H4-ST131 CTX- J. Jacobs); Institut National de Recherche are available from Africa, although M-15 extended-spectrum-β-lactamase– hospital-associated ESBL producers Biomédicale, Kinshasa, Democratic Republic producing Escherichia coli among have been described in Cameroon of the Congo (B. Miwanda, O. Lunguya- companion animals. J Antimicrob Che- and the Central African Republic Metila, J.J. Muyembe-Tamfum); Maastricht mother. 2010;65:651–60. http://dx.doi. org/10.1093/jac/dkq004 University Medical Center, Maastricht, the (6,7). ESBL-producing bacteria have 9. Warren RE, Ensor VM, O’Neill P, Butler been recovered from different sources Netherlands (E. Stobberingh); and Cliniques V, Taylor J, Nye K, et al. Imported chicken in the community, including food Universitaires Université Catholique de meat as a potential source of quinolone- and companion animals (8,9), and 1 Louvain de Mont-Godinne, Yvoir, Belgium resistant Escherichia coli producing ex- tended-spectrum β-lactamases in the UK. (Y. Glupczynski) recent study from India reported that J Antimicrob Chemother. 2008;61:504–8. a substantial number of tap water DOI: http://dx.doi.org/10.3201/eid1806.111214 http://dx.doi.org/10.1093/jac/dkm517 samples were contaminated with 10. Walsh TR, Weeks J, Livermore DM, carbapenemase bla producing Toleman MA. Dissemination of NDM-1 NDM-1 References positive bacteria in the New Delhi envi- organisms (10). ronment and its implications for human Kinshasa is the second-largest 1. Okeke IN, Laxminarayan R, Bhutta ZA, health: an environmental point prevalence city in sub-Saharan Africa. In Duse AG, Jenkins P, O’Brien TF, et al. study. Lancet Infect Dis. 2011;11:355–62. Antimicrobial resistance in developing 2008, of its estimated 8.7 million http://dx.doi.org/10.1016/S1473-3099 countries. Part I: recent trends and current (11)70059-7 inhabitants, only 46%had access status. Lancet Infect Dis. 2005;5:481–93. http://dx.doi.org/10.1016/S1473-3099 to safe drinking water, and 23% Address for correspondence: Hilde De Boeck, had access to improved sanitation (05)70189-4 2. Vlieghe E, Phoba MF, Tamfun JJ, Ja- Institute of Tropical Medicine, Nationalestraat facilities according to the World cobs J. Antibiotic resistance among 155, Antwerp, Belgium; email: hdeboeck@itg. Bank. Opportunistic pathogens in bacterial pathogens in central Africa: a be drinking water and poor sanitary review of the published literature be- conditions may increase the risk of tween 1955 and 2008. Int J Antimicrob Agents. 2009;34:295–303. http://dx.doi. developing infectious enterocolitis org/10.1016/j.ijantimicag.2009.04.015 for consumers, especially for those 3. Clinical and Laboratory Standards Institute. who are immunocompromised. Performance standards for antimicrobial It can eventually lead to chronic susceptibility testing: 21st informational supplement. CLSI document M100-S21. intestinal carriage of multidrug- Wayne (PA): The Institute; 2011. resistant organisms. The presence 4. Endimiani A, Hujer AM, Hujer KM, Novel of ESBL producers in the intestinal Gatta JA, Schriver AC, Jacobs MR, et Chlamydiaceae fl ora could also lead to horizontal al. Evaluation of a commercial microar- ray system for detection of SHV-, TEM-, Disease in Captive transfer of drug resistance genes from CTX-M-, and KPC-type -lactamase genes commensal fl ora to enteric pathogens. in gram-negative isolates. J Clin Micro- Salamanders This emergence of ESBL-producing biol. 2010;48:2618–22. http://dx.doi. bacteria and further community- org/10.1128/JCM.00568-10 To the Editor: Although 2 major 5. Bradford PA. Extended-spectrum associated infections poses a public diseases of amphibians, chytridiomy- β-lactamases in the 21st century: char- cosis and ranavirosis, have been rela- threat, especially in low-resource acterization, epidemiology, and detec- countries where surveillance is tion of this important resistance threat. tively well studied, enigmatic amphib- suboptimal and empiric treatment Clin Microbiol Rev. 2001;14:933–51. ian disease and death not attributable http://dx.doi.org/10.1128/CMR.14.4.933- of invasive infections often includes to any of the known amphibian diseas- 951.2001 es frequently occur (1). We describe third-generation cephalosporins. 6. Gangoué-Piéboji J, Bedenic B, Koul- la-Shiro S, Randegger C, Adiogo D, an apparently new disease in salaman- Hilde De Boeck, Ngassam P, et al. Extended-spectrum- ders that is associated with a novel ge- Berthe Miwanda, β-lactamase-producing Enterobacteria- nus within the family Chlamydiaceae. ceae in Yaounde, Cameroon. J Clin Mi- The salamanders seen in our clinic Octavie Lunguya-Metila, crobiol. 2005;43:3273–7. http://dx.doi. Jean-Jacques Muyembe-Tamfum, org/10.1128/JCM.43.7.3273-3277.2005 belonged to 1 of the following species: Ellen Stobberingh, 7. Frank T, Arlet G, Gautier V, Talar- Salamandra corsica, the Corsican fi re Youri Glupczynski, min A, Bercion R. Extended-spectrum salamander (5 animals from 1 collec- β-lactamase-producing Enterobacteria- tion); Neurergus crocatus, the yellow and Jan Jacobs ceae, Central African Republic. Emerg Infect Dis. 2006;12:863. http://dx.doi. spotted newt (11 animals from 3 col- org/10.3201/eid1205.050951 lections); or N. strauchii, Strauch’s 1020 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 18, No. 6, June 2012 LETTERS spotted newt (6 animals from 2 collec- in all 5 Corsican fi re salamanders; in mander species specimens (GenBank tions). All salamanders were captive 4/7, 1/3, and 1/1 yellow spotted newts; accession no. JN392919). These se- bred; housed in breeding colonies in and in 4/5 and 1/1 Strauch’s spotted quence differences point to the exis- private collections in Elsloo and Eind- newts. For taxon identifi cation, the tence of multiple strains with possible hoven, the Netherlands, Munich, Ger- 16S rRNA gene of the Chlamydiales host adaptation. many, and Brugge, Belgium; and 1–3 bacteria was amplifi ed and sequenced We determined the phylogenetic years of age. from the livers from 2 yellow spotted position of the novel taxon, named Disease was characterized by newts (1 from the collection in Elsloo, Candidatus Amphibiichlamydia sala- anorexia, lethargy, edema, and mark- the Netherlands and 1 from the collec- mandrae (online Technical Appendix), edly abnormal gait. Mortality rate tion in Munich, Germany), 1 Strauch’s identifi ed by using neighbor-joining was 100%. Animals in these collec- spotted newt, and 5 Corsican fi re sala- analysis with Kodon software (Ap- tions had no histories of disease. All manders. plied Maths, Sint-Martens-Latem, animals were in good nutritional con- The sequences shared >90% nt Belgium). The novel Chlamydiales dition. Necropsy did not yield any identity with the 16S rRNA gene of C. forms a distinct branch in the well- macroscopic lesions. All animals had abortus B577 (GenBank accession no. supported monophyletic clade with mild intestinal nematode or protozoan D85709) and therefore can be identi- the genera Chlamydia and Candidatus infections. Results of real-time PCRs fi ed as a member of the family Chla- Clavochlamydia salmonicola (family for iridoviruses in liver and skin (2) or mydiaceae (5). The closest 16S rRNA Chlamydiaceae) (Figure). Maximum Batrachochytrium dendrobatidis fun- similarity (92%) was observed with C. parsimony and unweighted pair group gus of skin (3) were negative for all psittaci strain CPX0308 (AB285329). with arithmetic mean analyses yielded animals. The sequence obtained from all spot- cladograms with the same topology We placed liver suspensions from ted newt species specimens was (results not shown). Previous reports of the dead salamanders on Columbia identical (GenBank accession no. members of the family Chlamydiaceae agar with 5% sheep blood and tryptic JN392920) but differed slightly (1%) in amphibians concerned species oc- soy agar and then incubated the sam- from that obtained from the fi re sala- curring in other vertebrate taxa as well: ples up to 14 days at 20°C. No con- sistent bacterial growth was observed. Histologic examination of 2 Corsican fi re salamanders and 1 yellow spot- ted newt revealed hepatitis in 1 of the Corsican fi re salamanders and the yel- low spotted newt. Hepatitis was char- acterized by high numbers of melano- macrophages and a marked infi ltration of granulocytic leukocytes. Immuno- histochemical staining for chlamydia (IMAGEN Chlamydia; Oxoid, Bas- ingstone, UK) showed cell-associated fl uorescently stained aggregates in liver tissue, suggestive of Chlamydi- ales bacteria. Transmission electron microscopic examination of the liver of a yellow spotted newt revealed intracellular inclusions containing particles matching the morphology of reticulate or elementary bodies of Chlamydiaceae (online Technical Ap- pendix, wwwnc.cdc.gov/EID/pdfs/11- 1137-Techapp.pdf). Figure. Topology of the novel amphibian Chlamydiaceae (Candidatus Amphibiichlamydia A PCR (4) to detect the 16S rRNA salamandrae) within the phylogenetic tree obtained by neighbor-joining and based on of all Chlamydiales bacteria, per- 16S rRNA gene data from representative species. Numbers show the percentage of times each branch was found in 1,000 bootstrap replicates. The tree has been rooted with formed on liver tissue samples from Verrucomicrobium spinosum as outgroup. Scale bar indicates nucleotide substitutions per all animals, yielded positive results site. Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 18, No. 6, June 2012 1021 LETTERS C. psittaci, C. pneumoniae, C. abortus, 2. Mao J, Hedrick RP, Chichar VB. Molecular Novel Variant and C. suis (6–10). To our knowledge, characterization, sequence analysis, and taxonomic position of newly isolated fi sh of Beilong this member of the family Chlamydi- iridoviruses. Virology. 1997;229:212–20. aceae has been seen in amphibians, but http://dx.doi.org/10.1006/viro.1996.8435 Paramyxovirus not in other vertebrate hosts.
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