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Francisella Novicida–Causing Two Samples of Blood Cultures from Peripheral Lines Bacteremia in a Woman from Thailand Who Was Receiving Chemotherapy for Ovarian Cancer

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Francisella Novicida–Causing Two Samples of Blood Cultures from Peripheral Lines Bacteremia in a Woman from Thailand Who Was Receiving Chemotherapy for Ovarian Cancer she was treated with lamivudine. A follow-up visit in early Emergence of September showed that her liver function biochemistry re- sults had returned to within normal limits. Chemotherapy Francisella with carboplastin and paclitaxel was then initiated. At the time of admission, 25 days after the start of novicida chemotherapy, the patient had fever (39oC), blood pressure 90/60 mm Hg, and pulse rate 75 beats/min. She also had Bacteremia, an episode of gastrointestinal hemorrhage with melena. It Thailand was believed that fever and gastrointestinal bleeding were complications from chemotherapy; thus, microbiologic Amornrut Leelaporn, Samaporn Yongyod, investigation was not promptly initiated. Abnormal labo- Sunee Limsrivanichakorn, Thitiya Yungyuen, ratory fi ndings included anemia (hemoglobin 80 g/L) and and Pattarachai Kiratisin leukocytosis with marked neutrophilia (Figure). Urine and stool cultures showed insignifi cant growth. We report isolation of Francisella novicida–causing Two samples of blood cultures from peripheral lines bacteremia in a woman from Thailand who was receiving chemotherapy for ovarian cancer. The organism was iso- were obtained using BacT/Alert FA bottles (bioMérieux, lated from blood cultures and identifi ed by 16S rDNA and Durham, NC, USA) on day 10 of hospital admission and PPIase gene analyses. Diagnosis and treatment were de- incubated in the continuous monitoring BacT/Alert 3D sys- layed due to unawareness of the disease in this region. tem (bioMérieux). Both blood culture bottles grew small pleomorphic gram-negative coccobacillus after incubation for 2 days. Samples from positive bottles were subcultured rancisella novicida, a rare human pathogen, has recent- onto 5% (vol/vol) sheep blood agar, MacConkey agar, and Fly been considered to be a subspecies of F. tularensis chocolate agar. A slow-growing bacterium was recovered on the basis of DNA similarity (1,2). The reservoir and on both blood agar and chocolate agar after 2-day incuba- transmission route of F. novicida were not clearly defi ned. o tion at 35 C with 5% CO2. The organism was negative for Since the fi rst isolation of F. novicida, to our knowledge, catalase and oxidase. Bacterial identifi cation was delayed only 5 patients with suspected infection have been reported because the organism was unidentifi able based on the con- (3–5). F. novicida, however, has neither been isolated nor ventional biochemical keys and the Vitek 2 system (bio- associated with human disease in Thailand. We report a Mérieux, Marcy L’Etoile, France). Using the API 20NE case of F. novicida infection in a Thai patient who was un- (bioMérieux), the organism was identifi ed as Mannheimia dergoing chemotherapy. haemolytica/Pasteurella trehalosi with good confi dence level (94.5% probability). The Study The identifi cation of this isolate was further determined In October 2007, a 37-year-old woman from Thailand by the 16S rDNA gene analysis that used screening prim- sought treatment at Siriraj Hospital (a 2,400-bed university ers SQE1 and SQE3 (6) and the 1,445-bp gene sequencing hospital in Bangkok, Thailand) with a history of fever for 1 procedure as described elsewhere (7). The DNA sequence week. She was a hairdresser residing in a suburban area of was analyzed by using the standard nucleotide-nucleotide Prachuap Khiri Khan, a southern province of Thailand. She BLAST algorithm (www.ncbi.nlm.nih.gov). The 16S denied history of blood transfusion, animal contact, and rDNA sequence of this isolate, called strain RB401, was travel abroad. She had not been aware of being bitten by most closely related (99.9% homology) to the novicida-like insects recently. There was no incidence of unusual animal death in the area in which she resided. Five months before 96.2% 3 seeking treatment, she received a diagnosis of advanced 40 40 /mm 3 C 91.3% o stage clear cell adenocarcinoma of the ovary with metas- 30 39 tasis to peritoneum, spleen, uterus, and multiple abdominal 90.2% 89.2% 89.0% 20 38 lymph nodes. Chemotherapy was planned. Initial labora- 92.0% Onset of Hospital Blood culture Hospital Patient tory screening showed increased liver enzyme levels and Temperature, 10 fever admission positive discharge died 37 abnormal hepatitis markers confi rming chronic active hep- Leukocyte count, x10 atitis B virus infection. Chemotherapy was delayed while 0 5 10 15 20 25 30 Course of illness, d Author affi liation: Siriraj Hospital, Mahidol University, Bangkok, Thailand Figure. Course of illness in a Thai patient with Francisella novicida bacteremia. Solid line, leukocyte count with percentage of DOI: 10.3201/eid1412.080435 neutrophils; dashed line, temperature. Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 14, No. 12, December 2008 1935 DISPATCHES subspecies of F. novicida strain 3523 (GenBank accession There is no clear explanation regarding route of ac- no. AY243028) and was deposited to the GenBank data- quisition and the pathogenic role of this organism. Addi- base under the accession no. EU365864. tionally, there is no evidence for human-to-human trans- Identifi cation of strain RB401 was further confi rmed mission. In our case, clinicians and microbiologists did not by analyzing DNA sequences of TUL4 and PPIase genes as suspect F. novicida infection. Given that our patient had previously described for strain 3523 (5). The fragment con- severe complicated underlying diseases while the poten- taining TUL4 amplicon (363 nt) from strain RB401 shared tially low-virulence organism was recovered, it was thus the highest similarity (98%) to the matching region from diffi cult to claim that the patient’s death was solely due to strain 3523. Comparing the 267-nt coding region sequence F. novicida infection. Notably, the 16S rDNA, TUL4, and of the 3′ end of PPIase gene nt 733259–733525 to the ge- PPIase sequences of our isolate are most closely related to nome sequence of F. novicida strain U112 (GenBank ac- the strain earlier reported from Australia (5). These areas cession no. CP000439) showed the highest similarity to all are not known to be endemic for F. novicida, and therefore, subspecies of F. tularensis, e.g., F. tularensis subsp. tularen- these strains represent the emergence of F. novicida in the sis strain WY96-3418 (GenBank accession no. CP000608), Asia-Pacifi c region. The earlier case-patient was success- and to F. novicida strain U112. The PPIase gene sequence fully treated with fl ucloxacillin and doxycycline, followed of strain 3523 was not available for the alignment. Howev- by dicloxacillin and doxycycline (5). er, the above fi ndings also supported that our isolate was F. F. novicida is not considered to have a fastidious novicida. The sequences of TUL4 and PPIase genes from growth requirement. The standard protocol of 5-day incu- strain RB401 were deposited to GenBank under accession bation for automated blood culture (9) is supposedly suf- nos. EU786119 and EU786120, respectively. fi cient for detection of F. novicida bacteremia. Identifi ca- Antimicrobial drug susceptibility testing was per- tion of F. novicida, however, is often diffi cult because the formed by disk diffusion method and interpreted based on bacterium can be easily misidentifi ed as a non-Francisella the CLSI criteria for Acinetobacter spp. (8). Results showed species or as a highly pathogenic F. tularensis. It was also that strain RB401 was susceptible to piperacillin/tazobac- indicated that this organism could not be detected by a di- tam, third-generation cephalosporins, cefepime, imipenem, rect fl uorescent antibody test used for the identifi cation of meropenem, aminoglycosides, fl uoroquinolones, and tetra- F. tularensis types A and B (5). As a preliminary result, our cycline, but resistant to co-trimoxazole. Due to the delay in 16S rDNA sequence was closely related to subspecies of F. organism identifi cation and lack of awareness of its signifi - tularensis, and thus doxycycline prophylaxis for the organ- cance, antimicrobial treatment (piperacillin/tazobactam 4.5 ism-exposed laboratory personnel could not be avoided. g intravenously every 8 h) was not initiated until day 19. There is high risk for laboratory-acquired tularemia when However, on day 21, the patient requested to be referred handling F. tularensis cultures (10). to her hometown hospital and died 2 days after referral. In retrospect, prophylactic treatment may have been The fi nal microbiology report was released 2 days after her unnecessary because our isolate proved to be F. novicida, death. The serologic test for antibodies against Francisella which is not known to cause laboratory-acquired infec- spp. was not available. All laboratory technicians who pro- tions. Rapid methods for Francisella species identifi ca- cessed culture and identifi ed the bacterium received doxy- tion are needed for better consideration of antimicrobial cycline (100 mg twice a day for 14 days) for prophylaxis, prophylaxis. Most F. tularensis strains were susceptible and none reported fever or abnormal symptoms. to aminoglycosides, quinolones, and tetracyclines (11). Our isolate was similarly susceptible to these agents, Conclusions which suggests that they may be considered as therapeu- F. novicida, often referred to as F. tularensis subsp. tic options for Francisella spp. Initial treatment with pip- novicida, is rarely attributed to human infection and is eracillin/tazobactam was based on the preliminary report not readily recognized in most clinical laboratories. Be- of gram-negative bacilli, but later antimicrobial suscepti- cause of the unreliable results of
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