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Home , Leptospira, Leptospiraceae, Leptospira noguchii

LETTERS

Leptospira noguchii previously isolated from animals such titer of 25 against saprophytic sero- as armadillo, toad, spiny , opossum, var Andamana by MAT. Both patients and Human and nutria, the least weasel (Mustela niva- were from the rural area of Pelotas. Animal , lis), cattle, and the oriental fi re-bellied Unfortunately, convalescent-phase se- Southern Brazil toad (Bombina orientalis) in Argen- rum samples were not obtained from tina, Peru, Panama, Barbados, Ni- these patients. To the Editor: Pathogenic lep- caragua, and the United States (1,6). A third isolate (Hook strain) was tospires, the causative agents of lep- Human leptospirosis associated with obtained from a male stray with tospirosis, exhibit wide phenotypic L. noguchii has been reported only in , lethargy, weight loss, disori- and genotypic variations. They are the United States, Peru, and Panama, entation, diarrhea, and vomiting. The currently classifi ed into 17 and with the isolation of strains Autum- animal died as a consequence of the >200 serovars (1,2). Most reported nalis Fort Bragg, Tarassovi Bac 1376, disease. The isolate was obtained from cases of leptospirosis in Brazil are of and Undesignated 2050, respectively a tissue culture. No temporal urban origin and caused by (1,6). The Fort Bragg strain was iso- or spatial relationship was found be- interrogans (3). Brazil underwent a lated during an outbreak among troops tween the 3 cases. dramatic demographic transformation at Fort Bragg, North Carolina. It was Serogrouping was performed by due to uncontrolled growth of urban identifi ed as the causative agent of an using a panel of rabbit antisera. Bo- centers during the last 60 years. Urban illness characterized by , head- nito, Cascata, and Hook strains were slums are sites of poor sanitation that ache, myalgia, and a pretibial rash— classifi ed as Autumnalis, Bataviae, favors rat-borne transmission of lep- Fort Bragg fever (7). We were not able and Australis, respectively. Sero- tospirosis among humans. Thus, this to obtain data regarding the other 2 groups were confi rmed by the strong may explain the major involvement of human isolates. and specifi c reaction of hyperim- serovar Copenhageni (L. interrogans). We report the isolation of 3 ad- mune sera against these isolates, with The predominance of L. interrogans ditional L. noguchii strains from Bra- the reference strains of the respec- is likely due to the underestimation of zil, including 2 from cases of human tive serogroups. Species identifi ca- rural cases of leptospirosis. leptospirosis. The fi rst isolate (Bonito tion was accomplished by sequenc- Pelotas is a coastal city in Rio strain) was obtained from the ing nearly the full length of the 16S Grande do Sul State, in southern Bra- culture of a 34-year-old man who rRNA gene, as previously described zil, with ≈400,000 inhabitants. This exhibited fever, headache, myalgia, (5). The sequences of the Hook, Cas- state has a typical temperate climate. hemorrhages, , abdominal cata, and Bonito strains were depos- However, the incidence of human pain, diarrhea, and vomiting. The pa- ited in GenBank under accession nos. leptospirosis is high (12.5/100,000 tient reported contact with , farm EU349494–EU349496. inhabitants in 2001) compared with animals, and before the onset of In addition, the rpoB gene se- the mean incidence in areas of Brazil illness. Laboratory tests at admission quence was determined and used for where tropical and subtropical cli- to the Hospital Santa Casa de Miser- further confi rmation of the species. mates predominate (3.5/100,000 in the icórdia, Pelotas, showed an elevated The rpoB sequence for the strains same year). Most cases in Rio Grande level of serum (total 21 mg/ Hook, Cascata, Bonito, and the L. do Sul State (69%) occur in rural areas dL, direct 16 mg/dL) and a slight in- noguchii reference strains were de- where the spatial distribution suggests crease in liver levels (alanine posited in GenBank under accession an association with areas of rice fi eld aminotransferase 2×, aspartate ami- nos. EU349497–EU349505. BLAST activities. Pelotas had an annual inci- notransferase 1.5× above reference (www.ncbi.nlm.nih.gov/blast/Blast. dence of >50 cases per 100,000 inhab- levels). An acute-phase serum sample cgi) alignment confi rmed the new iso- itants in 2001, which placed it among showed a titer of 25 against serovars lates as L. noguchii. The 16S rRNA the cities with the highest incidence of Autumnalis and Bratislava by micro- gene sequence was also used for tax- leptospirosis in southern Brazil (4). scopic agglutination test (MAT). onomic analysis of L. noguchii (Fig- Before 2007, pathogenic serovars The second isolate (Cascata ure). The topology-based dendrogram and strains in Brazilian collections in- strain) was obtained from the blood demonstrates sequence relatedness cluded the following species: L. san- culture of a 16-year-old boy who ex- among strains isolated in Pelotas and tarosai, L. interrogans, L. kirshneri, hibited headache, fever, fl ulike symp- the L. noguchii Autumnalis, Australis, and L. borgpetersenii. However, our toms, and myalgia. He reported previ- and Bataviae strains deposited in Gen- research group has recently reported ous contact with rats and dogs. The Bank (Figure). No molecular or sero- the isolation of L. noguchii in Brazil patient was not hospitalized, and an logic characterization at the serovar from sheep (5). This species had been acute-phase serum sample showed a level was performed.

Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 15, No. 4, April 2009 621 LETTERS

We report herein the occurrence the isolates was performed at the Gonçalo Éverton F. Silva, of L. noguchii species in southern Bra- Moniz Research Centre. Gustavo M. Cerqueira, zil. The 3 isolates obtained belong to Núbia Seyffert, This work was supported by Co- distinct serogroups. Information pre- Fabiana K. Seixas, ordenação de Aperfeiçoamento de Pessoal sented here places L. noguchii among Daiane D. Hartwig, de Nível Superior Foundation (Brazilian the prevalent Leptospira species that Daniel A. Athanazio, Government), Bio-Manguinhos, Oswaldo are able to cause human and animal Luciano S. Pinto, Cruz Foundation, Brazilian Ministry of leptospirosis in southern Brazil. Adriano Queiroz, Health (09224-7), the Brazilian National Albert I. Ko, Claudiomar S. Brod, Research Council (300.861/96-6), and the and Odir A. Dellagostin Isolation procedures and DNA se- National Institutes of Health (AI-052473, Author affi liations: Universidade Federal de quencing were conducted at the Federal TW-00919). Pelotas, Pelotas, Brazil (É.F. Silva, G.M. University of Pelotas. Serogrouping of Cerqueira, N. Seyffert, F.K. Seixas, D.D. Hartwig, L.S. Pinto, C.S. Brod, O.A. Della- gostin); Universidade Federal da Bahia, Salvador, Brazil (D.A. Athanazio); Funda- ção Oswaldo Cruz, Salvador (D.A. Athana- zio, A. Queiroz, A.I. Ko); and Weill Medical College of Cornell University, New York, New York, USA (A.I. Ko)

DOI: 10.3201/eid1504.071669

References

1. Brenner DJ, Kaufmann AF, Sulzer KR, Steigerwalt AG, Rogers FC, Weyant RS. Further determination of DNA relatedness between serogroups and serovars in the family with a proposal for sp. nov. and four new Leptospira genomospecies. Int J Syst Bacteriol. 1999;49:839–58. 2. Salaun L, Merien F, Gurianova S, Ba- ranton G, Picardeau M. Application of multilocus variable-number tandem- repeat analysis for molecular typing of the agent of leptospirosis. J Clin Micro- biol. 2006;44:3954–62. DOI: 10.1128/ JCM.00336-06 3. Pereira MM, Matsuo MG, Bauab AR, Vas- concelos SA, Moraes ZM, Baranton G, et al. A clonal subpopulation of sensu stricto is the major cause of leptospirosis outbreaks in Brazil. J Clin Microbiol. 2000;38:450–2. 4. Barcellos C, Lammerhirt CB, de Almeida MA, Santos E. Spatial distribution of lep- tospirosis in Rio Grande do Sul, Brazil: recovering the ecology of ecological stud- ies [in Portuguese].Cad Saude Publica. Figure. Dendogram constructed by using the neighbor-joining algorithm, based on a 2003;19:1283–92. DOI: 10.1590/S0102- 1,180-bp sequence of the 16S rRNA gene demonstrating the position of the Brazilian 311X2003000500007 strains (Bonito, Cascata, and Hook) within the species. This 5. Silva EF, Brod CS, Cerqueira GM, Bours- dendogram summarizes, by bootstrap-based topology, the evolutionary relationship cheidt D, Seyffert N, Queiroz A, et al. Iso- among L. noguchii strains. The bootstrap consensus values are indicated over each root. lation of Leptospira noguchii from sheep. The initial lowercase letters indicate the respective species to which each strain belongs: Vet Microbiol. 2007;121:144–9. DOI: i, L. interrogans; k, L. kirschneri; n, L. noguchii; b, L. borgpetersenii; w, L. weilii; s, L. 10.1016/j.vetmic.2006.11.010 santarosai; a, L. alexanderi; f, L. fainei; in, L. inadai; br, L. broomi; bif, L. bifl exa. (T) 6. Faine SB, Adler B, Bolin C, Perolat P. indicates the type-strain for each species. The GenBank accession number follows the Leptospira and leptospirosis, 2nd ed. Mel- strain identifi cation. bourne (Australia): MediSci; 1999.

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7. Fraser DW, Glosser JW, Francis DP, genomic island 1 (SGI1) were sur- 3. Ribot EM, Wierzba RK, Angulo FJ, Bar- Phillips CJ, Feeley JC, Sulzer CR. Lep- rounded by non–antimicrobial-drug rett TJ. Salmonella enterica Ty- tospirosis caused by serotype Fort-Bragg. phimurium DT104 isolated from humans, A suburban outbreak. Ann Intern Med. resistance DNA. This DNA is homol- United States, 1985, 1990, and 1995. 1973;79:786–9. ogous to DNA sequences in plasmids Emerg Infect Dis. 2002;8:387–91. PASPPFLO and pJA8122 (see Figure 4. Cloeckaert A, Baucheron S, Chaslus-Dan- Address for correspondence: Éverton F. Silva, 1 and Table 2 in reference 5) (5–7). In cla E. Nonenzymatic chloramphenicol resistance mediated by IncC plasmid R55 Universidade Federal de Pelotas, Centro de addition to antimicrobial drug resis- is encoded by a fl oR gene variant. Antimi- Biotecnologia, Campus Universitário, Cx. tance genes, PASPPFLO and pJA8122 crob Agents Chemother. 2001;45:2381–2. Postal 354, 96010-900. Pelotas, RS, Brazil; contain cloned DNA segments of in- DOI: 10.1128/AAC.45.8.2381-2382.2001 email: [email protected] digenous R plasmids found in V. dam- 5. Briggs CE, Fratamico PM. Molecular characterization of an resistance sela and V. anguillarum, respectively; gene cluster of Salmonella typhimurium these cloned DNA segments span DT104. Antimicrob Agents Chemother. sequences that extend beyond their 1999;43:846–9. fl orfenicol resistance and tetR/tetG 6. Kim E, Aoki T. Sequence analysis of the fl orfenicol resistance gene encoded in the genes (5–7). For example, the region transferable R-plasmid of a fi sh , of the fl orfenicol resistance gene in Pasteurella piscicida. Microbiol Immu- SGI1 contains 763 nt of the non–an- nol. 1996;40:665–9. Aquaculture timicrobial-drug resistance portion of 7. Zhao J, Aoki T. Nucleotide sequence anal- ysis of the class G resistance and Florfenicol the original V. damsela plasmid; the determinant from Vibrio anguillarum. Mi- Resistance in region of tetR/tetG contains 468 nt of crobiol Immunol. 1992;36:1051–60. the non–antimicrobial-drug resistance Salmonella enterica DNA segment of the P. piscicida plas- Address for correspondence: Felipe C. Cabello, Serovar mid (5–7). Department of Microbiology and Immunology, Typhimurium DT104 The presence of these non–anti- New York Medical College, Valhalla, NY microbial-drug resistance R plasmid 10595, USA; email: [email protected] To the Editor: In a letter re- DNA sequences in SGI1 constitutes cently published in Emerging Infec- a molecular signature that fi rmly es- tious Diseases, Smith (1) discussed tablishes the aquaculture origin of evidence that he mistakenly believes the fl orfenicol resistance and the tetR/ to undermine the hypothesis that the tetG genes in the S. enterica serovar fl orfenicol resistance gene present in Typhimurium DT104 strain studied by In Response: In his letter (1), Ca- some isolates of the epidemic Salmo- Briggs and Fratamico and in the SGI1 bello makes 2 observations regarding nella enterica serovar Typhimurium of other (5). These R plasmid the debate concerning the origin of the DT104 strain originated from a fl o- DNA sequences in SGI1 also confi rm fl oR gene in Salmonella enterica sero- rfenicol resistance plasmid present in direct or indirect horizontal gene trans- var Typhimurium DT104. The fi rst ob- Vibrio damsela (Pasteurella piscicida) fer between bacteria in the aquaculture servation is that the plasmid PASPP- that infected fi sh farms in Japan in the environment and S. enterica serovar FLO contained cloned segments of an 1990s (2). Smith correctly states that Typhimurium DT104 (5–7). indigenous Vibrio damsela plasmid. the fl orfenicol resistance gene was However, PASPPFLO is not the name present in S. enterica serovar Typh- Felipe C. Cabello of a plasmid but is the GenBank locus imurium DT104 strains isolated in the Author affi liation: New York Medical Col- identifi er associated with the sequence United States in 1985, before the gene lege, Valhalla, New York, USA (GenBank accession no. D37826) of was documented in V. damsela in Ja- a 3,745-bp region of the V. damsela DOI: 10.3201/eid1504.081171 pan (1,3). He is also correct in noting plasmid pSP92088 that contained pp- that this particular fl orfenicol resis- fl o (2,3). References tance gene was detected in a plasmid The second observation is that in Klebsiella pneumoniae in France in 1. Smith P. Aquaculture and fl orfenicol resis- sequences fl anking the fl oR gene in S. 1969 (1,4). tance in Salmonella enterica Typhimurium enterica serovar Typhimurium DT104 However, an earlier report by DT104. Emerg Infect Dis. 2008;14:1327–8. (GenBank accession no. AF071555) DOI: 10.3201/eid1412.080162 Briggs and Fratamico (5) clearly es- 2. Angulo FJ, Griffi n PM. Changes in an- are homologous to those fl anking the tablished that the fl orfenicol resistance timicrobial resistance in Salmonella en- pp-fl o gene sequenced from the V. genes and the tetracycline resistance terica serovar Typhimurium. Emerg Infect damsela plasmid pSP92088 (4). On genes tetG and tetR in the Salmonella Dis. 2000;6:436–8. the basis of this homology, he seems

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