(See the Major Article by Neil et al, on pages 10919 and the Editorial Commentary by Crump, on pages 11079.)
Background. Salmonella enterica serovar Typhi causes an estimated 22 million cases of typhoid fever and
216 000 deaths annually worldwide. We investigated an outbreak of unexplained febrile illnesses with neurologic
findings, determined to be typhoid fever, along the MalawiMozambique border.
Methods. The investigation included active surveillance, interviews, examinations of ill and convalescent
persons, medical chart reviews, and laboratory testing. Classification as a suspected case required fever and $1 other
finding (eg, headache or abdominal pain); a probable case required fever and a positive rapid immunoglobulin M
antibody test for typhoid (TUBEX TF); a confirmed case required isolation of Salmonella Typhi from blood or stool.
Isolates underwent antimicrobial susceptibility testing and subtyping by pulsed-field gel electrophoresis (PFGE).
Results. We identified 303 cases from 18 villages with onset during MarchNovember 2009; 214 were suspected,
43 were probable, and 46 were confirmed cases. Forty patients presented with focal neurologic abnormalities,
including a constellation of upper motor neuron signs (n 5 19), ataxia (n 5 22), and parkinsonism (n 5 8). Eleven
patients died. All 42 isolates tested were resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole;
4 were also resistant to nalidixic acid. Thirty-five of 42 isolates were indistinguishable by PFGE.
Conclusions. The unusual neurologic manifestations posed a diagnostic challenge that was resolved through
rapid typhoid antibody testing in the field and subsequent blood culture confirmation in the Malawi national
reference laboratory. Extending laboratory diagnostic capacity, including blood culture, to populations at risk for
typhoid fever in Africa will improve outbreak detection, response, and clinical treatment.
Typhoid fever is a systemic illness that often presents Typhi), which is transmitted by the fecal-oral route.
with fever, headache, and abdominal pain. The etiologic Multiple severe complications can occur, including
agent is Salmonella enterica serovar Typhi (Salmonella intestinal hemorrhage, intestinal perforation, hepatitis,
pneumonia, abscess, and neuropsychiatric abnormali-
ties [1]. An estimated 22 million cases and 216 000
Received 20 June 2011; accepted 13 October 2011; electronically published 22
February 2012.
deaths occur worldwide each year [2].
a
Present affiliation: New York State Department of Health, Albany. Typhoid fever is endemic in Malawi and Mozambique
Correspondence: Eric Mintz, MD, MPH, Division of Foodborne, Waterborne, and
Environmental Diseases, CDC, Mailstop C-09, 1600 Clifton Rd, Atlanta, GA 30333
[37]. Both are considered medium-incidence countries
(emintz@cdc.gov). with estimated rates of 10100 per 100 000 persons
Clinical Infectious Diseases 2012;54(8):11006 per year [2]. Multidrug-resistant strains of Salmonella
Published by Oxford University Press on behalf of the Infectious Diseases Society of
America 2012.
Typhi, defined as resistant to ampicillin, chloram-
DOI: 10.1093/cid/cis012 phenicol, and trimethoprim-sulfamethoxazole, have
Figure 1. Location of the typhoid outbreak in villages on the MalawiMozambique border, in the center of the circle, 2009.
Typhoid Fever With Neurologic Findings d CID 2012:54 (15 April) d 1101
capacity to collect and transport specimens to reference labo- and Escherichia coli by using presence-absence broth with
ratories was established, confirmatory blood and stool cultures 4-methylumbelliferyl-b-D-glucuronide (Hach Company,
were performed at the Community Health Study Unit, Malawis Loveland, Colorado).
national reference laboratory. Blood specimens were inoculated
to commercially prepared BD BACTEC or BBL Septi-Check Ethics
blood culture bottles according to manufacturers specifications This investigation was initiated and conducted in response to
for manual subculture (BD, Franklin Lakes, New Jersey). When an illness outbreak. Human subjects research designees at
available, stool and rectal swab specimens were collected for CDC determined that the activities constituted public health
culture. All specimens were transported to the reference labo- response and program evaluation rather than research. Verbal
ratory at times ranging from immediately after collection to consent for specimen collection was obtained from patients or
1 week, depending on availability of transportation. Specimens guardians.
were subcultured according to CDC standard protocols for
isolation of Salmonella Typhi from blood and stool specimens RESULTS
[16]. Isolates were biochemically confirmed to be Salmonella
and serotyped as Typhi by using specific antisera for Vi, O9, and Epidemiologic Investigation
Hd antigens at CDC. Antimicrobial susceptibility testing of A total of 303 persons with illness onset during 1 March13
Salmonella Typhi isolates using an automated broth micro- November 2009 met the case definition; 214 suspected, 43
dilution assay (Sensititre, Trek Diagnostics, Cleveland, Ohio) probable, and 46 confirmed cases were identified. The outbreak
and pulsed-field gel electrophoresis (PFGE) were also performed appeared to peak in September 2009 (Figure 2). The median age
at CDC [17]. of patients was 21 years (range, 181 years); 125 of 299 (42%)
patients with known age were aged 519 years. Overall, 166 of
Case Definition
299 (56%) patients in whom sex was known were female;
We defined a suspected case of typhoid fever as illness in
females outnumbered males in all but 2 age groups (Figure 3).
a resident of Neno District, Malawi, or Tsangano District,
Eighty-one persons (27%) were hospitalized, and 11 died
Mozambique, with onset on or after 1 March 2009, who pre-
(case fatality rate: 4%). Clinical data for the 303 cases are
sented with fever (subjective or with a documented temperature
displayed in Table 1.
$38C), and at least 1 of the following signs and symptoms:
Forty persons (13%) had objective, focal neurologic findings
gastrointestinal illness (abdominal pain, vomiting, diarrhea, or
documented in the medical chart or elicited on examination;
constipation), joint pain, muscle pain, headache, neck pain
27 (68%) of these patients were hospitalized; 5 (13%) died. An
or stiffness, difficulty walking or talking, arms held in flexion,
additional 27 persons had altered mental status but no focal
or mental status changes, with no alternative explanation for
neurologic findings. Twenty-six of the 40 persons with focal
the illness. A probable case required fever and a positive rapid
neurologic signs met criteria for a suspected case, 10 for
typhoid test (TUBEX TF), and a confirmed case required either
a probable case, and 4 for a confirmed case. Patients with focal
a blood culture yielding Salmonella Typhi, or fever and a stool
neurologic findings did not differ from those without neuro-
culture yielding Salmonella Typhi. Patients with positive
logic findings by age (P 5 .29) or sex (P 5 .68). The median age
malaria smears were counted as cases only if there was no
of patients with neurologic findings was 18 years (range, 357
response to an initial course of antimalarial medication.
years), and 53% were female. Neurologic signs and symptoms
A neurologic case was defined as illness in a patient with
among all hospitalized patients are displayed in Table 2.
objective, focal neurologic findings documented in the medical
Nineteen of the 40 persons (48%) with objective neurologic
chart or elicited on examination. The presence of altered mental
manifestations had a constellation of findings indicative of
status or dysarthria did not satisfy the definition of a neurologic
upper motor neuron dysfunction, defined as documentation of
case because in this setting, these findings could not reliably
at least 2 of the following findings: limb hyperreflexia, spas-
differentiate true neurologic deficits from symptoms referable
ticity, presence of Babinski sign, or sustained ankle clonus.
to severe systemic illness. Similarly, reports of hearing loss or
Other notable neurologic features included ataxia (22, 55%)
vertigo did not satisfy the definition of a neurologic case
and features of parkinsonism (gait instability, global slowness
because of the necessarily subjective nature of the complaints in
of movement, facial masking) (8, 20%). Subjective hearing loss
this setting.
(9, 23%) and vertigo (2, 5%) were noted among the 40 patients
Water Source Assessment with objective, focal neurologic findings and also among
Water samples from unprotected springs in 2 villages and from patients without objective findings (Table 1). Magnetic reso-
a semiprotected spring in 1 village, all of which were used as nance imaging of the brain and spinal cord was performed on
drinking water sources, were tested for total coliform bacteria 3 persons with focal neurologic findings at the time of the
scan. No focal abnormalities were present, although all 3 was negative for acute infection with all viruses tested. All
demonstrated generalized cortical and cerebellar atrophy viral cultures were negative. Autopsy specimens from a decedent
disproportionate to age. who had focal neurologic findings revealed only patchy ne-
crosis in the liver; central nervous system samples showed no
Laboratory Investigation significant histopathologic changes.
A total of 412 individual tests for viral, bacterial, parasitic, and Seventeen patients (8 neurologic) underwent cerebrospinal
rickettsial pathogens were performed on specimens taken at the fluid examination; all parameters tested were normal. Human
beginning of the investigation from persons meeting the case immunodeficiency virus serologic testing was performed on
definition (Supplementary Table). All pathogen-specific PCR, 14 patients (5 neurologic); serology was positive in 1 non-
random-primer PCR, and 16S ribosomal RNA sequencing neurologic patient. Malaria smears were performed in 44
results were negative for potential pathogens. Serologic testing persons (14 neurologic); 8 (18%) were positive (3 neurologic).
Typhoid Fever With Neurologic Findings d CID 2012:54 (15 April) d 1103
Table 2. Neurologic Signs and Symptoms Among Hospitalized were distinguished from the predominant pattern only by the
Typhoid Fever Patients (n 5 81), Neno District, Malawi, and second enzyme (BlnI); 2 differed by the first enzyme (XbaI) but
Tsangano District, Mozambique, 2009 not the second; 1 differed by both enzymes.
Typhoid Fever With Neurologic Findings d CID 2012:54 (15 April) d 1105
Potential conflicts of interest. A. P. received support for travel to meet- Salmonella, and Shigella for PulseNet. Foodborne Pathog Dis 2006;
ings for the study or for other purposes from the government of Malawi; 3:5967.
R. L. received salary support; D. Talkington reported royalties for an 18. Theobald L, Halpin J, Pecic G, et al. Diversity of Salmonella enterica
invention not related to this study and travel and expenses paid by CDC. serovar Typhi isolates collected from eastern African countries based
All authors have submitted the ICMJE Form for Disclosure of Potential on pulsed-field gel electrophoresis and antimicrobial susceptibility
Conflicts of Interest. Conflicts that the editors consider relevant to the testing. Poster presented at the International Conference on Emerging
content of the manuscript have been disclosed. Infectious Diseases (ICEID). Atlanta, Georgia, 2010.
19. Osler W. The principles and practice of medicine. New York:
References D Appleton and Co, 1892.
20. Osuntokun BO, Bademosi O, Ogunremi K, Wright SG. Neuropsychi-
1. Parry CM, Hien TT, Dougan G, White NJ, Farrar JJ. Typhoid fever. atric manifestations of typhoid fever in 959 patients. Arch Neurol 1972;
N Engl J Med 2002; 347:177082. 27:713.
2. Crump JA, Luby SP, Mintz ED. The global burden of typhoid fever. 21. Khosla SN, Srivastava SC, Gupta S. Neuro-psychiatric manifestations
Bull World Health Organ 2004; 82:34653. of typhoid. J Trop Med Hyg 1977; 80:958.
3. Archibald LK, McDonald LC, Nwanyanwu O, et al. A hospital-based 22. Sawhney IM, Prabhakar S, Dhand UK, Chopra JS. Acute cerebellar
prevalence survey of bloodstream infections in febrile patients in ataxia in enteric fever. Trans R Soc Trop Med Hyg 1986; 80:856.
Malawi: implications for diagnosis and therapy. J Infect Dis 2000; 23. Ali G, Rashid S, Kamli MA, Shah PA, Allaqaband GQ. Spectrum of
181:141420. neuropsychiatric complications in 791 cases of typhoid fever. Trop
4. Gordon MA, Graham SM, Walsh AL, et al. Epidemics of invasive Med Int Health 1997; 2:31418.
Salmonella enterica serovar enteritidis and S. enterica serovar Typhi- 24. Parmar RC, Bavdekar SB, Houilgol R, Muranjan MN. Nephritis and
murium infection associated with multidrug resistance among adults cerebellar ataxia: rare presenting features of enteric fever. J Postgrad
and children in Malawi. Clin Infect Dis 2008; 46:9639. Med 2000; 46:1846.
5. Walsh AL, Phiri AJ, Graham SM, Molyneux EM, Molyneux ME. 25. van Wolfswinkel ME, Lahri H, Wismans PJ, Petit PL, van Genderen PJ.
Bacteremia in febrile Malawian children: clinical and microbiologic Early small bowel perforation and cochleovestibular impairment as
features. Pediatr Infect Dis J 2000; 19:31218. rare complications of typhoid fever. Travel Med Infect Dis 2009;
6. Gordon MA, Walsh AL, Chaponda M, et al. Bacteraemia and mortality 7:2658.
among adult medical admissions in Malawipredominance of non- 26. Wadia RS, Ichaporia NR, Kiwalkar RS, Amin RB, Sardesai HV. Cere-
Typhi salmonellae and Streptococcus pneumoniae. J Infect 2001; 42: bellar ataxia in enteric fever. J Neurol Neurosur PS 1985; 48:6957.
449. 27. Thapa R, Banerjee P, Akhtar N, Jain TS. Transient dysautonomia and
7. Sigauque B, Roca A, Mandomando I, et al. Community-acquired cerebellitis in childhood enteric fever. J Child Neurol 2008; 23:10812.
bacteremia among children admitted to a rural hospital in Mozambi- 28. Bhunia R, Hutin Y, Ramakrishnan R, Pal N, Sen T, Murhekar M.
que. Pediatr Infect Dis J 2009; 28:10813. A typhoid fever outbreak in a slum of South Dumdum municipality,
8. Mills-Robertson F, Addy ME, Mensah P, Crupper SS. Molecular West Bengal, India, 2007: evidence for foodborne and waterborne
characterization of antibiotic resistance in clinical Salmonella Typhi transmission. BMC Public Health 2009; 9:11522.
isolated in Ghana. FEMS Microbiol Lett 2002; 215:24953. 29. Farooqui A, Khan A, Kazmi SU. Investigation of a community out-
9. Wasfy MO, Frenck R, Ismail TF, Mansour H, Malone JL, Mahoney FJ. break of typhoid fever associated with drinking water. BMC Public
Trends of multiple-drug resistance among Salmonella serotype Typhi Health 2009; 9:47681.
isolates during a 14-year period in Egypt. Clin Infect Dis 2002; 35: 30. Aypak A, C xelik AK, Aypak C, Cxikman O. Multidrug resistant typhoid
12658. fever outbreak in Ercek villageVan, Eastern Anatolia, Turkey: clinical
10. Kariuki S, Revathi G, Muyodi J, et al. Characterization of multidrug- profile, sensitivity patterns, and response to antimicrobials. Trop Doct
resistant typhoid outbreaks in Kenya. J Clin Microbiol 2004; 42: 2010; 40:1602.
147782. 31. Olsen SJ, Kafoa B, Win NS, et al. Restaurant-associated outbreak
11. Akinyemi KO, Smith SI, Bola Oyefolu AO, Coker AO. Multidrug re- of Salmonella Typhi in Nauru: an epidemiological and cost analysis.
sistance in Salmonella enterica serovar Typhi isolated from patients Epidemiol Infect 2001; 127:40512.
with typhoid fever complications in Lagos, Nigeria. Public Health 2005; 32. Neil K, Sodha S, Luswago L, et al. Outbreak of typhoid fever with high
119:3217. rate of intestinal perforations, Kasese District, Uganda20082009.
12. Muyembe-Tamfum JJ, Veyi J, Kaswa M, Lunguya O, Verhaegen J, Presented at the 58th Annual Meeting of the American Society of
Boelaert M. An outbreak of peritonitis caused by multidrug-resistant Tropical Medicine and Hygiene. Washington, DC, 2009.
Salmonella Typhi in Kinshasa, Democratic Republic of Congo. Travel 33. Nkemngu NJ, Asonganyi EDN, Njunda AL. Treatment failure in a ty-
Med Infect Dis 2009; 7:403. phoid patient infected with nalidixic acid resistant S. enterica serovar
13. Coovadia YM, Gathiram V, Bhamjee A, et al. An outbreak of mul- Typhi with reduced susceptibility to ciprofloxacin: a case report from
tiresistant Salmonella Typhi in South Africa. Q J Med 1992; 82: Cameroon. BMC Infect Dis 2005; 5:4951.
91100. 34. Parry CM, Threlfall EJ. Antimicrobial resistance in typhoidal and
14. Mandomando I, Sigauque B, Morais L, et al. Antimicrobial drug re- nontyphoidal salmonellae. Curr Opin Infect Dis 2008; 21:5318.
sistance trends of bacteremia isolates in a rural hospital in southern 35. Crump J, Kretsinger K, Gay K, et al. Clinical response and outcome of
Mozambique. Am J Trop Med Hyg 2010; 83:1527. infection with Salmonella enterica serotype Typhi with decreased suscep-
15. Feasy NA, Archer BN, Heyderman RS, et al. Typhoid fever and invasive tibility to fluoroquinolones: a United States FoodNet multicenter retro-
nontyphoid salmonellosis, Malawi and South Africa. Emerg Infect Dis spective cohort study. Antimicrob Agents Chemother 2008; 52:127884.
2010; 16:144851. 36. Lewis MD, Serichantalergs O, Pitarangsi C, et al. Typhoid fever:
16. World Health Organization. Manual for the laboratory identification a massive, single-point source, multidrug-resistant outbreak in Nepal.
and antimicrobial susceptibility testing of bacterial pathogens of Clin Infect Dis 2005; 40:55461.
public health importance in the developing world. Available at: http:// 37. Mermin JH, Villar R, Carpenter J, et al. A massive epidemic of
www.who.int/csr/resources/publications/drugresist/WHO_CDS_CSR_ multidrug-resistant typhoid fever in Tajikistan associated with con-
RMD_2003_6/en/. Accessed 11 February 2011. sumption of municipal water. J Infect Dis 1999; 179:141622.
17. Ribot E, Fair M, Gautom R, et al. Standardization of pulsed-field gel 38. World Health Organization. Typhoid vaccines: WHO position paper.
electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Wkly Epidemiol Rec 2008; 83:4960.