# Novel Extended-spectrum $\beta$-Lactamase in Shigella sonnei

Agnes Lefort, Guillaume Arlet, Olivier Join-Lambert, Marc Lecuit, Olivier Lortholary

► To cite this version:

Agnes Lefort, Guillaume Arlet, Olivier Join-Lambert, Marc Lecuit, Olivier Lortholary. Novel Extended-spectrum $\beta$-Lactamase in Shigella sonnei. Emerging Infectious Diseases, 2007, 13 (4), pp.653-654. 10.3201/eid1304.061160. pasteur-03204194

HAL Id: pasteur-03204194

https://pasteur.hal.science/pasteur-03204194

Submitted on 21 Apr 2021

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Distributed under a Creative Commons Attribution 4.0 International License
Acknowledgments

We dedicate this work to the late Andrew Spielman, our coauthor and beloved mentor. We thank J. Rivero, I. Pérez, M. Méndez, I. Matheus, M. Aguiar de Bracho, I. Carreño, J.M. Hernández, A. Nagy, A. Suarez, N. Moncada, M. Kilpatrick, E. Rodriguez, E. Marquez, E. Marian, B. Hernández; C. Rivero-Blanco, M. Azar, J. Rodriguez, H. Montañez, F. Alfonzo, and G. Rangel for their contributions to this study; and the Centro de Investigaciones Biomédicas, Universidad de Carabobo and the Cell Culture Core of Wadsworth Center, New York State Health Department, for their support. We also thank ProFauna and the National Institute of Parks in Venezuela for permission to obtain mosquito and bird samples.

This study was supported by grant AI45440 from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, and an International Collaborations in Infectious Disease Research opportunity pool grant.

Irene Bosch,* Flor Herrera,†

Juan-Carlos Navarro,‡

Miguel Lentino,§ Alan Dupuis,¶#

Joseph Maffei,¶# Matthew Jones,¶#

Ernesto Fernández,**

Nelson Pérez,†† Jorge Pérez-Emán,‡

Anthony Érico Guimarães,‡‡

Roberto Barrera,§§

Nereida Valero,¶¶ Johnny Ruiz,†

Glenda Velásquez,### Juán Martinez,‡

Guillermo Comach,†

Nicholas Komar,***

Andrew Spielman,††††

and Laura Kramer¶#

Agrícolas, Maracay, Venezuela; ‡‡Instituto
Oswaldo Cruz, Rio de Janeiro, Brazil;
§§Centers for Disease Control and
Prevention, San Juan, Puerto Rico, USA;
¶¶Universidad del Zulia, Maracaibo,
Venezuela; ##Ministerio de Salud Insalud,
Carabobo, Venezuela; ***Centers for
Disease Control and Prevention, Fort
Collins, Colorado, USA; and †††† Harvard
School of Public Health, Boston,
Massachusetts, USA

References

1. Hayes EB, Komar N, Nasci RS, Montgomery SP, O'Leary DR. Campbell GL Epidemiology and transmission dynamics of West Nile virus disease. Emerg Infect Dis. 2005;11:1167-73.

2. Komar N, Clark GG. West Nile virus activity in Latin America and the Caribbean. Rev Panam Salud Publica. 2006;19:112-7.

3. Morales-Betoulle ME, Morales H, Blitvich BJ, Powers AM, Davis EA, Klein R, et al. West Nile virus in horses, Guatemala. Emerg Infect Dis. 2006;12:1038-9.

4. Mattar S, Edwards E, Laguado J, Gonzalez M. Alvarez J, Komar N. West Nile virus antibodies in Colombian horses. Emerg Infect Dis. 2005;11:1497-8.

5. Morales MA, Barrandeguy M, Fabbri C, Garcia GB, Vissani A, Trono K, et al. West Nile virus isolation from equines in Argentina, 2006. Emerg Infect Dis. 2006;12:1559-61.

6. Ebel GD, Dupuis AP II, Nicholas D, Young D. Maffei J, Kramer LD. Detection by enzyme-linked immunosorbent assay of antibodies to West Nile virus in birds. Emerg Infect Dis. 2002;8:979-82.

7. Dupuis AP II, Marra PP, Kramer LD. Serologic evidence of West Nile virus transmission, Jamaica, West Indies. Emerg Infect Dis. 2003;9:860-3.

8. Figueiredo LT. The Brazilian flaviviruses. Microbes Infect. 2000;2:1643-9.

9. Kuno G, Chang GJ, Tsuchiya KR, Karabatsos N, Cropp CB. Phylogeny of the genus Flavivirus. J Virol. 1998;72:73-83.

10. Calisher CH, Monath TP. Karabatsos N, Trent DW. Arbovirus subtyping: applications to epidemiologic studies, availability of reagents, and testing services. Am J Epidemiol. 1981;114:619-31.

Novel Extended-spectrum β-Lactamase in *Shigella sonnei*

**To the Editor:** A 38-year-old French man with a history of chronic juvenile arthritis was referred to the Necker-Enfants Malades University hospital (Paris, France) with a dysenteric syndrome. The patient had returned the day before from a 1-month stay in Port-au-Prince, Haiti, where he spent most of his time in close contact with young children from an orphanage, most of whom had diarrhea. Clinical examination at admission showed fever (39°C), chills, diffuse abdominal pain, bloody diarrhea, and vomiting. The patient received ceftriaxone, which was stopped on day 4 because initial blood and stool cultures were negative for pathogens and clinical signs had completely resolved.

Ten days later, he reported the
recurrence of diarrhea without fever.
A novel stool culture grew *Shigella
sonnei*. An extended-spectrum β-lac-
tamase (ESBL) was detected by dou-
ble-disk synergy test; the isolate was
also resistant to aminoglycosides
(except amikacin), tetracycline, and
cotrimoxazole. The strain was suscep-
tible to fluoroquinolones and fos-
fomycin. It also appeared susceptible
to azithromycin (MIC 4 µg/mL),
although azithromycin MIC for
*Shigella* spp. should be interpreted
with caution (1). The patient was suc-
cessfully treated with azithromycin at
a dose of 500 mg/day for 5 days.
Azithromycin was preferred to fluoro-
quinolones to avoid the risk for
tendinopathy because of the patient's
history of chronic juvenile arthritis
and because this antimicrobial agent
was shown to be effective in the treat-
ment of shigellosis caused by mul-
tidrug-resistant strains (2).

To identify the molecular basis of
this ESBL, a series of PCR primers

Address for correspondence: Irene Bosch, Center for Infectious Disease and Vaccine Research, University of Massachusetts Medical School, 55 Lake Ave North, Worcester, MA 01655, USA: email: Irene.bosch@umassmed.edu

*University of Massachusetts Medical School, Worcester, Massachusetts, USA; †Universidad de Carabobo Biomed, Maracay, Venezuela; ‡Universidad Central de Venezuela, Caracas, Venezuela; §Coleccion Ornitológica Phelps, Caracas, Venezuela; ¶New York State Department of Health, Albany, New York, USA; #State University of New York at Albany, Albany, New York, USA; **Universidad Central de Venezuela, Maracay, Venezuela; ††Instituto Nacional de Investigaciones

<footnote n="1">¹Deceased.</footnote>
were used for detection of TEM-, SHV-, or CTX-M-type ESBL (3). Only the TEM PCR showed positive results. Sequencing of 2 independent PCR products showed a new allele (www.lahey.org/studies/temtable.asp). Analysis of the deduced amino acid sequence allowed characterization of TEM-137, derived from TEM-1 with 2 substitutions, Arg-16→Ser and Glu-240→Arg. This ESBL (and resistance to aminoglycosides and tetracyclines) was easily transferred to *Escherichia coli* J53-2 by conjugation.

MICs of β-lactams alone or in association with clavulanic acid, were determined by E-test, according to manufacturer's instructions (AB Biodisk, Solna, Sweden). High-level resistance to ceftazidime (MIC 32 µg/mL) and intermediate resistance to cefotaxime (MIC 8 µg/mL) were observed; the strain remained susceptible to cefepime and imipenem (MIC 0.5 and 0.25 µg/mL, respectively). Clavulanic acid did not restore susceptibility to ceftazidime (MIC 4 µg/mL) but did restore susceptibility to cefotaxime (MIC 0.5 µg/mL). With clavulanic acid, the MIC of cefepime was 0.06 µg/mL.

ESBL in *S. sonnei* is rare worldwide. In Argentina, a CTX-M-2 was found in an isolate of *S. sonnei* resistant to cefotaxime but not to ceftazidime (4). In South Korea, TEM-15, TEM-17, TEM-19, TEM-20, TEM-52, and CTX-M-14 were characterized in *S. sonnei* (5); TEM-52 and CTX-M-14 were also widely distributed, particularly in *Salmonella* spp. (6,7). In Turkey, an isolate of *S. sonnei* producing CTX-M-3 was reported (8). In Hong Kong, sequencing of 2 *S. sonnei* isolates showed the presence of CTX-M-14 and CTX-M-15 (9). Finally, in Bangladesh, 2 isolates of *S. sonnei* with a class A ESBL were reported; they were not characterized at the molecular level, but the resistance phenotypes suggested a CTX-M type (10).

In our case, little information on antimicrobial drug resistance could be obtained from Haiti because no systematic investigation on resistance in *Enterobacteriaceae* is performed. Nevertheless, the emergence of TEM-137 (GenBank accession no. AM286274) harbored by this imported *S. sonnei* isolate clearly demonstrates that ESBL-associated shigellosis has emerged in Haiti and that potentially large and severe shigellosis outbreaks could occur, for which the use of azithromycin could be beneficial, as illustrated in our patient. Because treating shigellosis is becoming problematic, it is essential to focus on prevention measures such as simple rules of personal hygiene that might drastically decrease the risk of transmission.

This study was supported in part by a grant from the European Community, contract LSHM-CT 2003-503335.

Agnes Lefort,*† Guillaume Arlet,‡ Olivier F. Join-Lambert,*†

Marc Lecuit,*† and Olivier Lortholary*†

*Hôpital Necker-Enfants Malades, Université Paris V, Paris, France; †Centre d'Infectiologie Necker-Pasteur, Paris, France; and ‡Unité de Formation et de Recherche de Médecine Pierre et Marie Curie, Université Paris VI, Paris, France

References

1. Jain SK, Gupta A, Glanz B, Dick J, Siberry GK. Antimicrobial-resistant *Shigella sonnei*: limited antimicrobial treatment options for children and challenges of interpreting in vitro azithromycin susceptibility. *Pediatr Infect Dis J* 2005;24:494-7.

2. Khan WA, Seas C, Dhar U, Salam MA, Bennish ML. Treatment of shigellosis: V. Comparison of azithromycin and ciprofloxacin. A double-blind, randomized, controlled trial. *Ann Intern Med* 1997;126:697-703.

3. Eckert C, Gautier V, Saladin-Allard M, Hidri N, Verdet C, Ould-Hocine Z, et al. Dissemination of CTX-M-type beta-lactamases among clinical isolates of *Enterobacteriaceae* in Paris, France.

Antimicrob Agents Chemother. 2004;48: 1249-55.

4. Radice M, Gonzelez C, Power P, Vidal MC, Gutkind G. Third-generation cephalosporin resistance in *Shigella sonnei*, Argentina. *Emerg Infect Dis*. 2001;7:442-3.

5. Kim S, Kim J, Kang Y, Park Y, Lee B. Occurrence of extended-spectrum beta-lactamases in members of the genus *Shigella* in the republic of Korea. *J Clin Microbiol*. 2004;42:5264-9.

6. Lee K, Yong D, Yum JH, Kim HH, Chong Y. Diversity of TEM-52 extended-spectrum β-lactamase-producing non-typhoidal *Salmonella* isolates in Korea. *J Antimicrob Chemother*. 2003;52:493-6.

7. Yong D, Lim YS, Yum JH, Lee H, Lee K, Kim EC, et al. Nosocomial outbreak of pediatric gastroenteritis caused by CTX-M-14-type extended-spectrum β-lactamase-producing strains of *Salmonella enterica* serovar London. *J Clin Microbiol*. 2005;43:3519-21.

8. Acikgoz ZC, Gulay Z, Bicmen M, Gocer S, Gamberzade S. CTX-M-3 extended-spectrum beta-lactamase in a *Shigella sonnei* clinical isolate: first report from Turkey. *Scand J Infect Dis*. 2003;35: 503-5.

9. Cheung TK, Chu YW, Tsang GK, Ngang JY, Hui IS, Kam KS. Emergence of CTX-M-type beta-lactam resistance in *Shigella* spp. in Hong Kong. *Int J Antimicrob Agents*. 2005;25:350-2.

10. Rahman M, Shoma S, Rashid H, Siddique AK, Nair GB, Sack DA. Extended-spectrum beta-lactamase-mediated third-generation cephalosporin resistance in *Shigella* isolates in Bangladesh. *J Antimicrob Chemother*. 2004;54:846-7.

Address for correspondence: Agnes Lefort, Service de Médecine Interne, Hôpital Beaujon, 100 Bd du Général Leclerc, 92110 Clichy, France; email: aglefort@yahoo.com

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