Blackwell Science, LtdOxford, UKEMIEnvironmental Microbiology 1462-2920Blackwell Publishing Ltd, 200359719729Review ArticleDiversity of the genus BurkholderiaT.

Coenye and P. Vandamme

Environmental Microbiology (2003) 5(9), 719–729 doi:10.1046/j.1462-2920.2003.00471.x

Minireview

Diversity and significance of Burkholderia species

occupying diverse ecological niches

Tom Coenye* and Peter Vandamme gene sequences, showing the positions of all of the
Laboratory of Microbiology, Ghent University, K. L. Burkholderia species and representatives of related gen-
Ledeganckstraat 35, B-9000 Gent, Belgium. era, is shown in Fig. 1.

Summary Diversity of the genus Burkholderia

Members of the genus Burkholderia are versatile The genus Burkholderia was created by Yabuuchi et al.
organisms that occupy a surprisingly wide range of (1992) to accommodate the former rRNA group II
ecological niches. These bacteria are exploited for pseudomonads, excluding Pseudomonas pickettii and
biocontrol, bioremediation and plant growth promo- Pseudomonas solanacearum, which were transferred to
tion purposes, but safety issues regarding human the genus Ralstonia (Yabuuchi et al., 1995). Traditionally,
infections, especially in cystic fibrosis patients, have Burkholderia species are known as plant pathogens and
not been solved. This minireview gives an overview of soil bacteria with two important exceptions, B. mallei
the taxonomic and ecological diversity of the genus and B. pseudomallei, which are primary pathogens for
with particular emphasis on strains belonging to the humans and animals. Our present knowledge on the nat-
Burkholderia cepacia complex and addresses the ural diversity of members of this genus indicates that the
important question whether ‘good’ and ‘bad’ strains range of interactions between these bacteria and their
are actually the same. hosts is more complex, diverse, and, often, contradictory.
The interactions of some species seem restricted to one
type of host, whereas others have a much wider host
The genus Burkholderia contains over 30 species, which
range. The type of interaction may be that of a pathogen
occupy remarkably diverse ecological niches, ranging
but can also be symbiotic, or both.
from contaminated soils to the respiratory tract of humans.
Burkholderia caryophylli, Burkholderia plantarii,
The Burkholderia cepacia complex is ubiquitous in nature
Burkholderia glumae and Burkholderia andropogonis are
and can be found in soil, water (including sea water), the
the species that are known as plant pathogenic Burkhold-
rhizosphere of plants, in humans and various animal spe-
eria species. Burkholderia caryophylli is pathogenic for
cies and in the hospital environment. Burkholderia cepa-
carnations (Dianthus caryophyllus) but also causes onion
cia complex isolates have been exploited for various
rot (Ballard et al., 1970; Palleroni, 1984). Burkholderia
purposes, including biological control of plant pathogens,
plantarii causes seedling blight of rice and forms the dis-
bioremediation of recalcitrant xenobiotics and plant
ease-causing substance tropolone, a non-benzoid aro-
growth promotion. Unfortunately, some Burkholderia spe-
matic compound with a seven-membered ring (Azegami
cies have been involved in human infections, and safety
et al., 1987; Urakami et al., 1994). Burkholderia glumae
issues regarding these human infections are hampering
causes rot of rice grains and seedlings and has emerged
the wide-spread biotechnological applications. The pur-
during the past two decades as the most important bac-
pose of this minireview is to give an overview of the
terial pathogen of rice in Japan, Korea and Taiwan (Goto
remarkable diversity of the genus Burkholderia and to
and Ohata, 1956; Kurita and Tabei, 1967; Uematsu et al.,
shed some light on the important question whether or not
1976; Urakami et al., 1994). Burkholderia andropogonis
clinical and environmental B. cepacia complex isolates
is the causative agent of stripe disease of sorghum
are the same. A phylogenetic tree based on 16S rRNA
(Andropogon sp.) and leaf spot of velvet bean (Stizolo-
bium deeringianum). Two different pathovars (B. andro-
pogonis pv. andropogonis and pv. stizolobii) have been
Received 19 February, 2003; accepted 4 April, 2003. *For correspon-
dence. E-mail Tom.Coenye@UGent.be; Tel. (+32) 9 2645114; Fax described (Palleroni, 1984). Pseudomonas woodsii, an
(+32) 9 2645092. important pathogen of carnation (Smith, 1911), was
© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd
720 T. Coenye and P. Vandamme
recently shown to be a junior synonym of B. andropogonis
(Coenye et al., 2001a).
The majority of Burkholderia species however, are
known as soil bacteria, which exhibit different types of
non-pathogenic interactions with plants. The ecological
role of species like Burkholderia glathei, Burkholderia
graminis, Burkholderia phenazinium, Burkholderia car-
ibensis, Burkholderia caledonica, Burkholderia hospita,
Burkholderia terricola and Burkholderia sacchari is largely
unknown. Burkholderia glathei was proposed for strains
isolated from fossil lateritic soils in Germany (Zolg and
Ottow, 1975). Burkholderia phenazinium strains were iso-
lated from soil and were characterized by the production
of iodinin (Bell and Turner, 1973; Viallard et al., 1998).
Strains belonging to B. graminis were first isolated by
Viallard et al. (1998) from the rhizosphere of wheat, corn
and pasture grasses during a field survey of French and
Australian soils. Burkholderia caribensis was described by
Achouak et al. (1999) for a group of exopolysaccharide-
© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology, 5, 719–729
Fig. 1. 16S rDNA based phylogenetic tree
showing the positions of all of the Burkholderia
species and representatives of related genera.
Scalebar indicates 5% sequence dissimilarity.
producing strains isolated from a vertisol on the island of
Martinique. These bacteria constituted the dominant cul-
tivable population in this soil. B. caledonica strains were
isolated from the rhizosphere of different plants in Scot-
land (Coenye et al., 2001b). Some of these soil-borne
Burkholderia species were detected in several soil types
as transconjugants that acquired the plasmids pJP4 or
pEMT1 (containing genes required for the degradation of
2,4-dichlorophenoxyacetic acid) following inoculation of a
donor strain (Newby et al., 2000; Goris et al., 2002). A
more detailed study of some of these transconjugants
entailed the description of B. hospita and B. terricola
(Goris et al., 2002). Both species were isolated as pJP4
or pEMT1 containing transconjugants of agricultural soil
in Belgium; the latter has also been isolated from soil
samples in Scotland, Italy and Spain (P. Vandamme,
unpubl. obs.). The name B. sacchari was proposed for a
single strain recovered from the soil of a sugar-cane plan-
tation in Brazil (Brämer et al., 2001). This organism can

produce polyhydroxyalkanoates from a wide range of car-
bon sources (Brämer et al., 2001; 2002).
Several Burkholderia species have developed intimate
beneficial interactions with plants and colonize roots,
stems and leaves. The ability to fix atmospheric nitrogen
has been established in several burkholderias including
named species like Burkholderia vietnamiensis (see
below) and Burkholderia kururiensis [the name B. kuru-
riensis was proposed for a single strain isolated from an
acquifer polluted with trichloroethylene (Zhang et al.,
2000)] (Gillis et al., 1995; Magalhaes Cruz et al., 2001;
Estrada-De Los Santos et al., 2001), tentatively named
species like ‘Burkholderia tropicalis’ and ‘Burkholderia
brasilensis’, and unnamed strains probably representing
several novel species (Estrada-De Los Santos et al.,
2001). ‘Burkholderia tropicalis’ and ‘B. brasilensis’ have
not been formally described but strains of these putative
novel species have been recovered from various parts of
banana and pineapple plants in South-America. Burkhold-
eria tuberum and Burkholderia phymatum are novel spe-
cies (Vandamme et al., 2002) reported to be capable of
nitrogen fixation and nodulation in tropical legume plants
(Moulin et al., 2001). The former species was isolated
from root nodules of Aspalathus carnosa in South Africa;
the latter from root nodules of Machaerium lunatum in
French Guiana. The capacity to induce nodule formation
was reported for two B. caribensis and one B. dolosa (B.
cepacia genomovar VI, Vermis et al., 2003) isolate as well
(Moulin et al., 2001). However, recent experiments with
this B. dolosa isolate revealed that the nodA and nif genes
could not longer be detected (C. Boivin-Masson, pers.
comm.) suggesting that the isolate lost this capacity or
that the original observation was based on experimental
error. Although all of these species can be considered
endosymbionts, they are all culturable on agar slants in
normal laboratory conditions. Plant endosymbionts
belonging to the genus Burkholderia, which remain uncul-
tured despite numerous attempts have been reported in
several studies. Endosymbionts of leaf galls of Psychotria
kirkii were classified as Candidatus Burkholderia kirkii
(Van Oevelen et al., 2002). Uncultured bacterial endosym-
bionts of the arbuscular mycorrhizal fungi belonging to the
Gigasporaceae (Bianciotto et al., 1996; 2000) were
recently formally classified as Candidatus Glomeribacter
gigasporarum (Bianciotto et al., 2003). These bacteria are
close relatives of the genus Burkholderia and were also
reported to contain putative nif genes (Minerdi et al.,
2001). Most remarkable, Van Borm et al. (2002) recently
showed that the microflora inhabiting a pouch-shaped
organ at the junction of the midgut and the intestine of
Tetraponera ants partially consists of Burkholderia spe-
cies, which are most likely involved in the oxidative recy-
cling of nitrogen-rich metabolic waste. These organisms
are phylogenetically closely related to B. fungorum and B.
© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology, 5, 719–729
Diversity of the genus Burkholderia 721
caledonica but have so far not been cultured and their
exact taxonomic position remains uncertain.
Interactions between burkholderias and humans or ani-
mals are traditionally known for B. mallei and B.
pseudomallei. The former causes glanders in horses,
mules and donkeys and was one of the first biological
weapons of the 20th century, being used by Germany
during World War I (Wheelis, 1998). Glanders in humans
is acquired from infected animals or by contact with organ-
isms causing glanders via ingestion or inhalation (Palle-
roni, 1984). Burkholderia pseudomallei is the aetiological
agent of melioidosis in humans and animals, and is
endemic to South-east Asia, Northern Australia and tem-
perate regions that border the equator (Dance, 1991).
Humans can be infected by soil contamination of skin
abrasions, ingestion or inhalation (Dance, 2000). Sporad-
ically the organism is acquired via person-to-person or
animal-to-person spread (Dance, 2000). Burkholderia
pseudomallei is a saprophytic organism that routinely can
be isolated from environmental niches like water, moist
soil and rice paddies (Brook et al., 1997). These two spe-
cies were shown to represent a single genomic species
by DNA–DNA hybridization criteria (Rogul et al., 1970).
However, the major differences in their biochemical activ-
ities and in the clinical symptoms and epidemiology justify
this two-species concept. Recently, several environmental
B. pseudomallei-like organisms were formally classified
as Burkholderia thailandensis (Brett et al., 1998). In con-
trast to B. pseudomallei, B. thailandensis strains are not
correlated with human disease and are avirulent in the
Syrian golden hamster animal model (Brett et al., 1997).
One of the B. thailandensis isolates which was obtained
from a surface soil along a roadside in Thailand, was later
reclassified as Burkholderia ubonensis (Yabuuchi et al.,
2000). Phylogenetically, the latter species belongs to the
B. cepacia complex (see below, Fig. 1).
Finally, several Burkholderia species occupy multiple
niches, may have both pathogenic and symbiotic interac-
tions with plants, and have become known as opportunis-
tic pathogens in humans. Although they are not
considered important pathogens for the normal human
population, some are considered serious threats for spe-
cific patient groups such as cystic fibrosis patients. These
species include all Burkholderia cepacia complex bacte-
ria, Burkholderia gladioli and Burkholderia fungorum.
Burkholderia gladioli strains have been isolated from
decayed onions, Gladiolus sp., Iris sp. and rice, for which
this species is believed to be pathogenic, and from various
human clinical sources (Palleroni, 1984; Miyagawa and
Kimura, 1989; Ura et al., 1996). Burkholderia gladioli con-
sists of two pathovars (pv. gladioli and pv. allicola) as it
appeared that it comprised organisms with different
pathogenic capacities (Palleroni, 1984). Strains belonging
to Burkholderia cocovenenans (also known as
722 T. Coenye and P. Vandamme
‘Pseudomonas farinofermentans’) produce toxoflavin and
bongkrekic acid and are responsible for cases of food
poisoning caused by consumption of fermented corn flour
(Zhao et al., 1995). Burkholderia cocovenenans was
shown to be a junior synonym of B. gladioli (Coenye et al.,
1999); the latter having nomenclatural priority. Similarly,
also Pseudomonas antimicrobica [a name proposed for a
single isolate that was antagonistic to a wide range of
phytopathogenic bacteria and fungi (Attafuah and Brad-
bury, 1989; Walker et al., 1996)] was demonstrated to be
a junior synonym of B. gladioli (Coenye et al., 2000).
Burkholderia fungorum too has been recovered from a
wide range of ecological niches, including root nodules of
plants, fungi, and clinical specimens of humans and ani-
mals (Coenye et al., 2001b; P. Vandamme, unpubl. obs.).
The Burkholderia cepacia complex
Like many other burkholderias, B. cepacia was originally
known as a plant pathogenic pseudomonad. Pseudomo-
nas cepacia was described by Burkholder in 1950 as the
causative agent of bacterial rot of onion bulbs, causing a
disease called sour skin (Burkholder, 1950). In subse-
quent years however, the group of human opportunistic
pathogens known as ‘eugonic oxidisers group 1¢ and,
later, Pseudomonas kingii, and the environmental organ-
ism known as Pseudomonas multivorans (Morris and
Roberts, 1959; Stanier et al., 1966; Jonsson, 1970), were
shown to represent the same species (Ballard et al., 1970;
Snell et al., 1972; Samuels et al., 1973; Sinsabaugh and
Howard, 1975). During the past decade, international col-
laborative polyphasic-taxonomic studies (Coenye et al.,
2001c) have demonstrated that strains identified as B.
cepacia actually represent a complex of closely related
species or genomovars. This group, collectively referred
to as the B. cepacia complex, comprises at least nine
species (Table 1) sharing a high degree of 16S rDNA (98–
100%) and recA (94–95%) sequence similarity, and mod-
erate levels of DNA–DNA hybridization (30–50%) (Van-
damme et al., 1997; 2000; 2002; 2003; Coenye et al.,
Table 1. Overview of the Burkholderia cepacia complex.
Name Other designation
B. cepacia B. cepacia genomovar I
B. multivorans B. cepacia genomovar II
B. cenocepacia B. cepacia genomovar III
B. stabilis B. cepacia genomovar IV
B. vietnamiensis B. cepacia genomovar V
B. dolosa B. cepacia genomovar VI
B. ambifaria B. cepacia genomovar VII
B. anthina B. cepacia genomovar VIII
B. pyrrocinia B. cepacia genomovar IX
B. ubonensisa
a. The taxonomic position of B. ubonensis within the B. cepacia complex has not been fully established
© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology, 5, 719–729
2001d; 2001e). Apart from B. cepacia, also genomovars
V and IX were identified as established Burkholderia spe-
cies, i.e. B. vietnamiensis (Gillis et al., 1995) and B. pyr-
rocinia (Imanaka et al. 1965) respectively. Isolates from B.
multivorans (genomovar II), B. cenocepacia (genomovar
III) and B. stabilis (genomovar IV) were present in histor-
ical P. multivorans or P. kingii strain collections (Stanier
et al., 1966; Ballard et al., 1970; Samuels et al., 1973) but
were only considered representing novel species with the
application of species delineation standards as currently
defined (Wayne et al., 1987). Until now, strains from
genomovars VI (B. dolosa), VII (B. ambifaria) and VIII (B.
anthina) have not been found in historical collections.
All of these B. cepacia complex species have been
isolated from environmental and human clinical sources.
Animal infections caused by B. cepacia complex have
been reported (Berriatua et al., 2001) but in general, its
distribution in animal species and infection therein, is not
well-documented. Burkholderia cepacia complex bacteria
are universal contaminants of cosmetic and pharmaceu-
tical solutions (for reviews see Pankhurst and Philpott-
Howard, 1996; Jimenez, 2001) and can cause contami-
nation of water supplies (see for example Koenig and
Pierson, 1997), sterile solutions (like disinfectant solu-
tions, mouth wash and anaesthetics), and disposable
equipment leading to nosocomial infections and pseu-
doepidemics (for a review see Coenye and LiPuma,
2003). Burkholderia cepacia complex organisms have
also been recovered from unpasteurised bovine milk
(Moore et al., 2001) and from gelatine, a product of animal
origin often used in the food industry (De Clerck and De
Vos, 2002). In humans B. cepacia complex bacteria have
been associated with a wide variety of infections, most
often in patients with an underlying disabling illness
(including persons with chronic granulomatous disease).
However, severe community acquired infections (including
endocarditic and pneumonia) have been reported as well
(for a review see Coenye and LiPuma, 2003). More impor-
tantly, patients with cystic fibrosis (CF) are particularly
susceptible to infections caused by these bacteria.
References
Vandamme et al. (1997)
Vandamme et al. (1997)
Vandamme et al. (1997; 2003)
Vandamme et al. (1997; 2000)
Gillis et al. (1995); Vandamme et al. (1997)
Coenye et al. (2001d); Vermis et al. (2003)
Coenye et al. (2001e)
Vandamme et al. (2002)
Vandamme et al. (2002)
Yabuuchi et al. (2000)

B. cepacia complex and CF
Cystic fibrosis is the most frequent hereditary disease in
Caucasian populations, affecting approximately 1 in 2570
live births (Rosenstein and Zeitlin, 1998). Cystic fibrosis
symptoms are caused by a mutation in the cystic fibrosis
transmembrane conductance regulator (CFTR) gene, a
cAMP dependent transepithelial chloride channel. The
consequences of mutant CFTR protein are complex but
the resultant altered epithelial surface liquid in some way
predisposes the respiratory tract of affected persons to
bacterial infections (Dinwiddie, 2000). Airway infections in
CF patients are characterized by intercurrent acute exac-
erbations with fever, weight loss, increased cough, change
in volume, colour or appearance of sputum, increased
respiratory rate, dyspnea and appearance of infiltrates on
chest radiographs (Gilligan, 1991). Typically, periods of
relative well-being are followed by episodes of these pul-
monary exacerbations, which result in a progressive dete-
rioration of the lung function. The improvement of
antimicrobial therapy has contributed significantly to the
increased median survival age but chronic pulmonary
infection is still the leading cause of death among CF
patients (Liou et al., 2001). Common bacterial pathogens
in young CF patients include Staphylococcus aureus and
Haemophilus influenzae. Pseudomonas aeruginosa infec-
tion becomes more common during adolescence and by
adulthood almost 80% of CF patients are infected with this
organism (Lyczak et al., 2002).
Several reports on the recovery of bacteria now known
as B. cepacia complex from CF patients appeared in the
late 1970s and early 1980s (LiPuma, 1998). The first
detailed description of its clinical significance was pub-
lished by Isles et al. (1984), who also described the so-
called ‘cepacia syndrome’, a progressive respiratory fail-
ure with necrotising pneumonia that affected some but not
all patients. Similar increases in incidence were subse-
quently noted in other North-American and European CF
centres (reviewed in Coenye and LiPuma, 2003). From
these initial studies, several risk factors for B. cepacia
complex acquisition could be identified, including age,
severity of underlying lung disease, use of aminoglycoside
antibiotics, previous hospitalization and the presence of a
B. cepacia complex colonized sibling. Clustering of cases
in some centres and the decrease of colonization follow-
ing segregation of colonized and non-colonized patients
in other centres suggested that B. cepacia complex could
be transmitted between CF patients; since then numerous
reports have provided epidemiologic and genotyping evi-
dence for interpatient spread (reviewed in Coenye and
LiPuma, 2003). Compared to P. aeruginosa, B. cepacia
complex infects only a small proportion of CF patients but
nevertheless has a significant impact on survival (Liou
et al., 2001). In addition, it has been shown that B. cepacia
© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology, 5, 719–729
Diversity of the genus Burkholderia 723
complex colonized CF patients have a much poorer out-
come following lung transplantation (the only therapeutic
option for succesfull intermediate term survival for CF
patients with end-stage pulmonary disease) than their
non-infected counterparts (Aris et al., 2001; LiPuma,
2001).
Biocontrol, bioremediation and plant-growth-promotion
applications
The biological control of plant diseases, insects and
nematodes by microorganisms (both bacteria and fungi)
has been proposed as an alternative or a supplement to
chemical pesticides and the use of introduced biological
control could have enormous ecological and economical
benefits (Parke and Gurian-Sherman, 2001; Gerhardson,
2002). The two traditional approaches used for biological
control of soil-borne plant pathogens in the field have
been (i) crop rotation, to allow time for residential antag-
onists to ‘sanitize’ the soil or for propagules of specialized
pathogens to die, and (ii) the addition of organic amend-
ments to soil to stimulate residential antagonists. How-
ever, the greatest progress towards biological control of
soil-borne plant pathogens has been made with antago-
nists introduced with the planting material, i.e. biological
control with plant-associated microorganisms (Cook,
1990). Burkholderia cepacia complex strains can be
used to control seedling and root diseases in vitro and in
field tests, and can replace chemical alternatives. Field
tests have shown that such strains can colonize the
rhizosphere of several crops, including corn, maize, rice,
pea, sunflower and radish, and thereby significantly
increases the crop yield, even in the absence of patho-
gens (Parke et al., 1991; McLoughlin et al., 1992; Bowers
and Parke, 1993; Hebbar et al., 1998; Tran Van et al.,
2000). One of the most-studied biocontrol strains is B.
ambifaria strain AMMDT. It was isolated from the rhizo-
sphere of peas (Pisum sativum) grown in a Aphanomy-
ces root rot nursery in Wisconsin (USA) in 1985. It has
activity against Pythium aphanidermatum (responsible for
pre- and post-emergence damping-off in peas) and Aph-
anomyces euteiches (responsible for root rot in peas)
(Parke, 1990; Parke et al., 1991; Bowers and Parke,
1993; King and Parke, 1993; Heungens and Parke, 2000;
2001; Parke and Gurian-Sherman, 2001).
The exceptional metabolic versatility of B. cepacia com-
plex bacteria and strains belonging to other Burkholderia
species can also be used for bioremediation purposes.
Constituents of crude oils [including polycyclic aromatic
hydrocarbon (PHA) compounds], herbicides (including
2,4-dichlorophenoxyacetic acid and 2,4,5-trichlorophe-
noxyacetic acid, the principal component of Agent
Orange), TCE and ether derivatives used as gasoline
additives can be degraded by Burkholderia isolates (Kil-
724 T. Coenye and P. Vandamme
bane et al., 1982; Folsom et al., 1990; Krumme et al., can have serious implications for potential biotechnologi-
1993; Bhat et al., 1994). Well-characterized biodegrada- cal applications.
tive strains include G4 (Nelson et al., 1987; Folsom et al.,
1990; Shields et al., 1991; Leahy et al., 1996; McClay
Prevalence of members of the B. cepacia complex in CF
et al., 1996; Massol-Deya et al., 1997) and CRE-7 (Muel-
patients and in the environment
ler et al., 1996), which both belong to the B. cepacia
complex. Several Burkholderia strains with potential appli- All nine B. cepacia complex species have been identified
cations in bioremediation are extremely well-studied with in CF sputum cultures, but several recent studies indicate
regard to their biotechnological potential but are taxonom- that they are unequally represented. Surveys form the US
ically poorly characterized. For example, strain NF100 (LiPuma et al., 2001), Canada (Speert et al., 2002), Italy
(Hayatsu et al., 2000) and strain LB400 (Haddock et al., (Agodi et al., 2001) and Belgium (De Boeck and P. Van-
1993; Seeger et al., 1995; Billingsley et al., 1997; Master damme, unpubl. obs.) indicated that, although there are
and Mohn, 1998) are phylogenetically closely related to national differences, most patients are infected with B.
B. glathei and B. fungorum, respectively, but their exact cenocepacia or B. multivorans (Table 2).
taxonomic status remains to be determined. An overview There are much less systematic studies regarding the
of Burkholderia strains capable of degrading recalcitrant distribution of the B. cepacia complex species in environ-
xenobiotics can be found in the Biodegradative Strain mental samples. Balandreau et al. (2001) recovered 22 B.
Database (Urbance et al., 2003; http://bsd.cme.msu.edu/ cepacia complex isolates from the rhizosphere of wheat
bsd/index.html). and maize and from roots and shoots of wheat and lupine,
grown in France and Australia. Of these, 21 belonged to
B. cenocepacia, whereas a single isolate belonged to B.
Identification and misidentification of B. cepacia complex
ambifaria. Fiore et al. (2001) recovered 120 B. cepacia
isolates
complex isolates from maize rhizosphere. The majority
Identification of members of the B. cepacia complex is not belonged to B. cepacia, B. cenocepacia and B. ambifaria.
straighforward (see for example Shelly et al., 2000; Henry In a similar study, Bevivino et al. (2002) recovered 75
et al., 2001; Brisse et al., 2002). In a large-scale study, isolates from maize rhizosphere. Only B. cenocepacia
McMenamin et al. (2000) recovered 1051 isolates from (53.4%), B. ambifaria (37.3%), B. pyrrocinia (8.0%) and
608 CF patients receiving care in 115 treatment cities in B. cepacia (1.3%) were isolated. During a study of urban
91 centres in the USA. Of the 770 isolates referred as B. and suburban soils in three large cities in the USA, Miller
cepacia complex, 11% were misidentified. On the other and co-workers isolated B. cepacia complex isolates from
hand, of the 281 isolates received as unidentified or ‘not 14 (15%) of 91 sampled sites. Sixty-eight B. cepacia com-
B. cepacia complex’, 36% actually were B. cepacia com- plex isolates were recovered, mainly belonging to B. amb-
plex. Organisms that have been misidentified as B. cepa- ifaria and B. pyrrocinia (Miller et al., 2001; 2002). 127 B.
cia complex include P. aeruginosa, Stenotrophomonas cepacia complex isolates were recovered from agricultural
maltophilia, Ralstonia pickettii, Achromobacter xylosoxi- soils in the USA by Gonzalez et al. (2001). The majority
dans, Bordetella hinzii, Brevundimonas sp., Chryseobac- belonged to B. ambifaria (42.5%), B. cepacia (24.4%), B.
terium sp. and members of the Enterobacteriaceae. In cenocepacia (22.8%) and B. pyrrocinia (10.2%). Thus far,
addition, among isolates misidentified as B. cepacia com- overall, B. cepacia, B. cenocepacia, B. ambifaria and B.
plex, a variety of novel species mainly belonging to the b- pyrrocinia dominate in soil samples, whereas the other
Proteobacteria have been described. These comprise genomovars are rarely found. Although the knowledge
several novel Burkholderia species (including B. fun-
gorum, B. caledonica, B. terricola and B. hospita), novel
Ralstonia species (including Ralstonia gilardii, Ralstonia Table 2. Prevalence of Burkholderia cepacia complex organisms in
mannitolilytica and Ralstonia taiwanensis) and members CF specimens (in percentage of infected patients).
of the novel genera Pandoraea (Pandoraea apista, Pan-
USA Canada Italy Belgium
doraea pnomenusa, Pandoraea norimbergensis, Pando- Genomovar (n = 606) (n = 445) (n = 62) (n = 33)
raea pulmonicola and Pandoraea sputorum) and
I 2.6 0.2 4.8 –
Inquilinus (Inquilinus limosus). The clinical significance of II 37.8 9.7 4.8 57.8
these novel species is mostly unclear. Anecdotal evidence III 50.0 82.9 72.6 27.3
indicates that several are capable of chronic colonization IV 0.2 3.8 3.2 12.1
V 5.1 1.6 – 3.0
of the CF lung and/or person-to-person transmission. The VI 2.0 – – –
erroneous identification of an organism as B. cepacia VII 0.7 – – –
complex has serious medical, social and psychological VIII 0.2 – – –
IX 0.2 – – –
implications for infected CF patients and misidentification

© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology, 5, 719–729

about the distribution of the different genomovars in envi-
ronmental samples is restricted and although misidentifi-
cation is common, it is obvious that the same species that
occur in the environment, colonize and infect CF patients,
albeit to different degrees.
Is the natural environment the reservoir for B. cepacia
complex infections in CF patients?
Indirect evidence for the speculation that the environment
serves as a reservoir for acquisition of novel B. cepacia
complex species comes from the observation that infec-
tion control measures (including segregation of patients)
have reduced but not eliminated new infections (often
with isolates showing novel fingerprint types). Direct evi-
dence was obtained from genotyping studies, in which
clinical and environmental isolates were compared using
state-of-the-art molecular fingerprinting techniques.
Govan et al. (2000) showed that the B. cepacia type
strain ATCC 25416T.(isolated from rotting onions in the
1940s) was isolated from sputum of a CF patient in the
UK. LiPuma et al (2002) reported that B. cenocepacia
strain PHDC recovered from most CF patients in the mid-
Atlantic region of the USA could also be isolated from
agricultural soils. These findings, although anecdotal,
show that human isolates are not necessarily distinct
from environmental ones.
Concluding remarks
Although phylogenetically well-defined, the genus
Burkholderia is functionally a remarkably diverse genus.
The molecular and physiological background of this diver-
sity and adaptability are largely unknown. In addition, little
is known about factors that determine virulence, which
implies that only correct identification provides a first basis
for risk assessment and infection control. The genomes
of most Burkholderia species (including B. cepacia com-
plex species, B. glumae, B. glathei, B. gladioli and
Burkholderia sp. LB400) consist of two to four circular
replicons (Cheng and Lessie, 1994; Rodley et al., 1995;
Lessie et al., 1996; Wigley and Burton, 2000; Parke and
Gurian-Sherman, 2001). There appears to be a tremen-
dous variation in genome size among these phylogeneti-
cally closely related organisms ranging from 4.7 Mb to
more than 9 Mb. In addition, several Burkholderia species
harbour an extensive array of insertion sequences (Lessie
et al., 1996; Tyler et al., 1996) which may be involved in
genomic rearrangements and regulation of gene expres-
sion. The genomes of strains of several Burkholderia spe-
cies (including B. cenocepacia, B. pseudomallei and B.
mallei) have been completely sequenced and other
sequencing projects are in progress (Burkholderia sp.
LB400) or are planned for the near future (B. vietnamien-
© 2003 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology, 5, 719–729
Diversity of the genus Burkholderia 725
sis G4) (for an overview see http://igweb.integratedge-
nomics.com/GOLD/). It can be expected that knowledge
derived from these genome sequencing projects will allow
us to gain further insights into functional diversity, evolu-
tion and pathogenicity mechanism. This may provide the
scientific basis for important decisions regarding infection
control and the biotechnological use of Burkholderia
strains.
Acknowledgements
T. C. and P. V. are indebted to the Fund for Scientific
Research – Flanders (Belgium) for a position as postdoctoral
fellow and research grants respectively. T.C. also acknowl-
edges the support from the Belgian Federal Government
(Federal Office for Scientific, Technical and Cultural Affairs).
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