S H O R T C O M M U N I C AT I O N

Bartonella quintana in head louse nits

Emmanouil Angelakis, Jean-Marc Rolain, Didier Raoult & Philippe Brouqui
Faculté de Médecine et de Pharmacie, Université de la Méditerranée, URMITE UMR 6236, CNRS-IRD, Marseille, France

Correspondence: Philippe Brouqui, Faculté Abstract

de Médecine et de Pharmacie, Université de la
The body louse is the principal vector of Bartonella quintana, the causative

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Méditerranée, URMITE UMR 6236, CNRS-IRD,
27 Bd Jean Moulin, 13385 Marseille cedex 05,
France. Tel.: 133 491 38 55 17; fax: 133 491
83 03 90; e-mail:
IMMUNOLOGY & MEDICAL MICROBIOLOGY
organism of trench fever, but B. quintana DNA has also been detected in adult
head lice. Because there are no characteristics that distinguish the body louse from
the head louse, we decided to test head louse nits collected from a homeless man

philippe.brouqui@univmed.fr
Received 23 February 2011; revised 21 March
2011; accepted 23 March 2011.
Final version published online 5 May 2011.
DOI:10.1111/j.1574-695X.2011.00804.x
for the presence of B. quintana DNA. All of the sampled nits tested positive by real-
time PCR, and intergenic spacer region (ITS) gene sequences shared 100%
similarity to the corresponding ITS fragment of the genome of B. quintana. The
role of the head louse in the maintenance and transmission of B. quintana remains
to be determined.

Editor: Ake Forsberg

Keywords
Bartonella quintana; head louse; homeless; nits.

Head and body lice have been recognized as human parasites
for thousands of years (Light et al., 2008). Head lice live on
the scalp and lay eggs at the base of hair shafts, whereas body
lice live in the clothes on the surface of the body and lay their
eggs on clothing. The body louse is medically relevant as this
organism can transmit life-threatening bacteria to humans.
Entre
Among these bacteria are Rickettsia prowazekii, which causes
epidemic typhus, Bartonella quintana, the agent of trench
fever, and Borrelia recurrentis, the louse-borne relapsing
fever spirochete (Brouqui, 2011). With the exception of a
comprimento da tíbia
few phenotypic characteristics, such as the length of the tibia
on the second pair of legs, studies of the morphological
characters and primary endosymbionts of head and body
lice suggest that these two types of lice are conspecific
(Sasaki-Fukatsu et al., 2006). Moreover, genetic analysis has
not been able to show any differences between these two
subspecies; all molecular data available at this time show no
reciprocal monophyly between head and body lice and
indicate that no known species concept would recognize
these louse morphotypes as separate species (Light et al.,
2008). A study by Takano-Lee et al. (2003) demonstrated
criado através
that head lice can be reared successfully in vitro through a
complete life cycle. It has also been proposed that body louse
heterozygosity would be retained and would transform into
the head louse phenotype if the organism were relocated to
the head or reared under head louse conditions (Levene &
Dobzhansky, 1959). In both the homeless of developed
countries and in Nepalese children, dual infestation by both
body and head lice was found in up to 50% of the cases
examined, and under these circumstances, it is impossible to
differentiate between the head and the body lice (Brouqui,
2011). Finally, with the current state of knowledge, head lice
are defined by their location on the head and by the
attachment of their eggs (called nits) to the base of the hair
shaft, while body lice are defined by their location in the
folds of clothes. Head lice infestation occurs most often in
school children, but has also been reported in the homeless
and other impoverished populations (Sasaki et al., 2006).
Although Fournier and colleagues successfully identified B.
quintana and R. prowazekii in body lice collected from
people living in poor conditions, such as the homeless and
refugees (Badiaga et al., 2008), their earlier attempts to
identify the presence of B. quintana and R. prowazekii in
143 head lice collected from school children across eight
countries were not successful (Fournier et al., 2002). The
first evidence that B. quintana might infect head lice was
shown by Sasaki et al. (2006), who detected the bacterial
DNA in both head and body lice from two heavily infested
Nepalese children who were living on the streets or in slum
areas. Bonilla et al., (2009) reported the detection of
B. quintana DNA in the head lice of a homeless individual
from San Francisco, CA, without any known concurrent

FEMS Immunology & Medical Microbiology  c 2011 Federation of European Microbiological Societies FEMS Immunol Med Microbiol 62 (2011) 244–246
Published by Blackwell Publishing Ltd. No claim to original French government works
Bartonella quintana in head lice
Fig. 1. Nits collected from the hair of a homeless individual. Living larvae
within the nit sheathes and one larvae hatching (white arrow) are shown.
The photograph was taken using a Nikon light microscope at a
magnification of  20.
body louse infestation. Because the body and the head louse
are phenotypically indistinguishable, we decided to test head
louse nits for B. quintana DNA.
Since 2000, we have collaborated with homeless shelters
in Marseille, France, to investigate arthropod-borne diseases
in this socially displaced and deprived population (Badiaga
et al., 2008; Brouqui, 2011). The study was approved by our
ethical committee under No. 10-005. We regularly visited
the shelters and looked for body lice. During this survey, we
met a homeless male who presented with a massive head lice
infestation. While vagabond’s leukomelanoderma, a hall-
mark of past body louse infestation, was noted, no presence
of evolving scratching lesions was recorded. A conscientious
clinical examination revealed neither adult head nor adult
body lice on the removed clothes. Unfortunately, a blood
sample was not collected at the time of sampling head louse
nits. After obtaining the patient’s consent, we shaved the
patient and found only head louse nits. Some nits were
found about 3–3.5 cm from the hair follicle. Most nits were
full of larvae, and some had begun hatching (Fig. 1). DNA was
extracted from three sets of pooled nits using a QIAamp tissue
kit (Qiagen, Hilden, Germany). The DNA was used as a
template in a real-time PCR assay targeting a portion of the
Bartonella 16S–23S intergenic spacer region (ITS), as de-
scribed previously (Fournier et al., 2002), and two specific B.
quintana genes (fabF3 and yopP) that encode for 3-oxoacyl-
[acyl-carrier-protein] synthase and a hypothetical intracellular
effector, respectively. The reactions were performed with the
following primer sequences: (Bqui05300F/FabF3/5 0 -GCT-
GGC-CTT-GCT-CTT-GAT-GA-3 0 , Bqui05300R/FabF3/5 0 -
GCT-ACT-CTG-CGT-GCC-TTG-GA-30 , Bqui11580F/yopP/
5 0 -TAA-ACC-TCG-GGG-GAA-GCA-GA-3 0 , Bqui11580R/
yopP/50 -TTT-CGT-CCT-CAA-CCC-CAT-CA-3 0 ) and probes
(Bqui05300P/FabF3/6FAM-TGC-AGC-AGG-TGG-AGG-AGA-
FEMS Immunol Med Microbiol 62 (2011) 244–246
245
ACG-TG-TAMRA, Bqui11580P/yopP/6FAM-CGT-TGC-CGA-
CAA-GAC-GTC-CTT-GC-TAMRA). The specificity of these
encoding genes was verified in silico as well as on a panel of 14
different Bartonella species (data not shown). All three sets were
positive by real-time PCR. Amplicons were then purified using
the QIAquick Spin PCR purification kit (Qiagen, Courtaboeuf,
France) and sequenced on an ABI 3100 automated sequencer
(Perking Elmer, Courtaboeuf, France) using the dRhodamine
Terminator cycle-sequencing ready reaction kit (PE Applied
Biosystems, Les Ulis, France), according to the manufacturer’s
instructions. Sequences obtained after sequencing of the ITS
gene shared 100% similarity to the corresponding ITS fragment
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of the genome of B. quintana (Toulouse strain) (GenBank
accession number NC_005955). To cultivate B. quintana, a
fourth set of pooled nits was triturated in brain–heart medium
before inoculation onto Columbia 5% sheep blood agar plates
(BioMerieux, Marcy l’Etoile, France). Plates were placed in
polyethylene bags and incubated at 37 1C in 5% CO2 (Genbag
CO2 system; BioMerieux) (La Scola & Raoult, 1999). This assay
was repeated independently three times, and the agar plates were
examined weekly. After 1 month, no evidence of growth was
observed.
As the speed of hair growth is roughly 1.25 cm month1,
we can estimate that the head louse infestation in the
examined homeless individual occurred at least 3 months
back; however, because larvae were apparent through the
nit’s sheaths, the infestation appeared to be ongoing (Fig. 1).
To avoid the potential of misinterpretation of the PCR data
due to laboratory contamination, three different genes were
assessed, and all of them were successfully detected. The
source of the Bartonella DNA could have been the larvae of
the louse, the nit’s sheath or the cement deposit from the
accessory glands of an infected female head louse. DNA
contamination of the hair by means of body louse feces
containing B. quintana is unlikely, as no adult body lice were
found on the homeless individual’s clothes, and no scratch-
ing lesions were noted, indicating that the individual was
not currently infested with body lice.
We were not able to cultivate B. quintana from nits or
larvae. This result suggests that either the number of living
microorganisms in the tested pool was too small to be
isolated by culturing or that only DNA, but no living
organisms, was present in the nits.
Interestingly, all attempts to detect B. quintana in the
head lice of school children have failed (Fournier et al.,
2002). Bartonella quintana DNA has been detected exclu-
sively in head lice collected from impoverished populations
such as the homeless or Nepalese children living in slums or
on the streets, who are usually infested by both head and
body lice (Sasaki et al., 2006; Bonilla et al., 2009). In
alcoholic homeless individuals, B quintana bacteremia can
be prolonged for weeks (Foucault et al., 2006), and under
these circumstances, one could hypothesize that head lice
FEMS Immunology & Medical Microbiology  c 2011 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. No claim to original French government works
246
might be contaminated by blood that is infected with
B. quintana. Nonetheless, the role of the head louse in the
maintenance and transmission of B. quintana remains to be
determined.
Acknowledgements
This study was supported by CNRS UMR 6236. None of the
authors have a conflict of interest relevant to this study.
Statement
This article has been edited in English by the American
Journal Expert under No. WN29KQY3.
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