Genus

Proteobacteria/Betaproteobacteria/Hydrogenophilales/Hydrogenophilaceae/

Thiobacillus
Beijerinck 1904b, 597AL
..........................................................................................................................................................................................
Donovan P. Kelly, University of Warwick, Department of Biological Sciences, Coventry CV4 7AL, United Kingdom
Ann P. Wood, Microbiology Research Group, Kings College, London, Division of Life Sciences, Franklin-Wilkins Building,
150 Stamford Street, London SE1 8WA, United Kingdom
Erko S. Stackebrandt, Deutsche Sammlung von Mikroorganismen und Zellkulturen, GmbH, and GBF, Forschung GmbH2,
Mascheroder Weg 1b, Braunschweig D-38124, Germany

Thi.o.ba.cillus. Gr. n. thios sulfur; L n bacillus a small rod;
M. L. masc n. Thiobacillus sulfur rodlet.
Small, Gram-negative, rod-shaped cells (0.30.5
0.94.0 m). Some species are motile by means
of polar flagella. No resting stages known. Energy
is derived by the oxidation of one or more reduced
sulfur compounds, including sulfides, sulfur, thiosul-
fate, polythionates, and thiocyanate. Sulfate is the
end product of sulfur-compound oxidation, but sul-
fur, sulfite, and polythionates may be accumulated by
most species, sometimes transiently. All species can fix
carbon dioxide by means of the BensonCalvin cycle
and are capable of autotrophic growth; some species
are obligately chemolithotrophic, while others are
chemoorganotrophic. The genus currently includes
obligate aerobes and facultative denitrifiers. Optimal
pH of 28 with optimal temperature of 2843 C. Dis-
tribution is seemingly ubiquitous in marine, freshwater,
and soil environments, especially where oxidizable sul-
fur is abundant (e.g., sulfur springs, sulfide minerals,
sulfur deposits, sewage treatment areas, and sources
of sulfur gases, such as sediments or anaerobic soils
releasing H2 S).
The mol% G + C of the DNA is: 6267.
Type species: Thiobacillus thioparus Beijerinck 1904b,
597.
..................................................................................
......................................................................................................................................................................................................
Small, Gram-negative, rod-shaped cells (0.30.5 0.94.0 m).
Some species are motile by means of polar flagella. No rest-
ing stages known. Energy is derived by the oxidation of
one or more reduced sulfur compounds, including sul-
fides, sulfur, thiosulfate, polythionates, and thiocyanate.
Sulfate is the end product of sulfur-compound oxidation,
but sulfur, sulfite, and polythionates may be accumulated
by most species, sometimes transiently. All species can fix
carbon dioxide by means of the BensonCalvin cycle and are
capable of autotrophic growth; some species are obligately
chemolithotrophic, while others are chemoorganotrophic.
The genus currently includes obligate aerobes and facultative
denitrifiers. Optimal pH of 28 with optimal temperature
of 2843 C. Distribution is seemingly ubiquitous in marine,
freshwater, and soil environments, especially where oxidiz-
able sulfur is abundant (e.g., sulfur springs, sulfide minerals,
sulfur deposits, sewage treatment areas, and sources of sulfur
gases, such as sediments or anaerobic soils releasing H2 S).
The mol% G + C of the DNA is: 6267.
Type species: Thiobacillus thioparus Beijerinck 1904b, 597.
Number of validated species: 6
Further descriptive information
The chemolithotrophic, sulfur-compound-oxidizing, Gram-
negative, rod-shaped members of the genera and species
described (Table 1) are usually 0.5 1.04.0 m in size,
occurring singly, in pairs or in short chains; some are motile
by means of single polar flagella; some possess pili and other

Bergeys Manual of Systematics of Archaea and Bacteria, Online 2015 Bergeys Manual Trust. This article is 2005 Bergeys Manual Trust.
DOI: 10.1002/9781118960608.gbm00969. Published by John Wiley & Sons, Inc., in association with Bergeys Manual Trust.
 2 Bergeys Manual of Systematics of Archaea and Bacteria

TABLE 1. Differentiation of the genera of chemolithotrophic, sulfur-oxidizing, rod-shaped bacteria

Character Thiobacillus Acidiphilium Acidithiobacillus Halothiobacillus Paracoccus Starkeya Thermithiobacillus Thiomonas

Obligate + + + +
chemolithoautotropha
Heterotrophic growth on +b +c + d
defined media
Mol% G + C of DNA 6267 6368 5264 5667 6371 6768 6667 6167
Class:
Alphaproteobacteria + + +
Betaproteobacteria + +
Gammaproteobacteria + + +
Respiratory ubiquinone Q-8 Q-10 Q-8 Q-8 Q-10 Q-10 Q-8 Q-8
Facultative denitrification +e +c
Optimal temperature ( C) 2843 2537 3045 2830 2537 2530 4345 3050
Optimal pH 6.88.0f 3.03.5 2.03.5 6.58.0 6.59.0 7 6.87.5 5.26.0g
Halophilic or halotolerant +
Contains photosynthetic +
reaction centers
a With inorganic sulfur compounds as sole energy substrates.
b Most species of the genus are not chemolithoautotrophic.
c Not all species exhibit all these features, and only P. denitrificans, P. versutus, and P. pantotrophus are facultative, sulfur-oxidizing

chemolithoautotrophs.
d Most strains will grow on complex rich media, and some grow best mixotrophically with thiosulfate plus organic supplements.
e Denitrification to dinitrogen in one species only: T. denitrificans.
f T. plumbophilus grows only between pH 4.06.5.
g Optimum for T. cuprinus is pH 3.04.0.

specialized surface features. All can obtain energy from the
oxidation of reduced inorganic sulfur compounds, and, in
most cases, elemental sulfur. Some species are obligately
chemolithotrophic and autotrophic, others are faculta-
tively heterotrophic, and, in some strains, optimal growth
occurs mixotrophically. Both electron-transport-dependent
and substrate-level phosphorylation occur during sulfur-
compound oxidation, and reduction of NAD(P) requires
an energy-dependent flow of electrons from cytochrome
c (or b). Carbon dioxide fixation occurs mainly by means
of the BensonCalvin cycle, with some fixation occurring
through pyruvate or phosphoenolpyruvate carboxylation.
The obligately autotrophic species possess an incomplete
tricarboxylic acid cycle (lacking 2-oxoglutarate dehydroge-
nase), which is used as a biosynthetic horseshoe pathway
via oxaloacetate or succinate and 2-oxoglutarate, respectively,
down each arm of the horseshoe. Some species contain
characteristic plasmids, and some have been shown to be
susceptible to introduction of Pseudomonas plasmids. Little
work on the genetics of Thiobacillus spp. is available, but it is
known that autotrophic and drug-resistant mutants can be
obtained and that mutations that decrease autotrophic effi-
ciency can be induced. No species is known to be pathogenic.
Thiobacillus spp. are ubiquitous, with the facultative species
occurring in soil, freshwater, and marine environments as
heterotrophs or mixotrophs. Sulfur-compound-oxidizing
species have been isolated from Arctic, temperate, and trop-
ical waters, soil, salt marshes, freshwater lakes, rivers, canals,
hot springs, sulfur-rich mine or acid-mine wastewaters and
other environments where oxidizable sulfur compounds
occur naturally or anthropogenically.
Enrichment and isolation procedures
Most of the species can be isolated from natural habitats
by the use of mineral media containing elemental sulfur or
thiosulfate as the energy-yielding substrate. Use of media
of different pH will assist in differential selection of the
neutrophilic and acidophilic species, use of acid ferrous
sulfate medium will frequently select for Acidithiobacillus
ferrooxidans, and use of anaerobic thiosulfate medium (pH 7)
supplemented with nitrate will select for T. denitrificans.

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Bergeys Manual of Systematics of Archaea and Bacteria
A procedure has been described for the enrichment
of facultatively autotrophic, mixotrophic strains, using a
continuous-flow chemostat with both organic and inorganic
substrates (Gottschal and Kuenen, 1980). This provides a
means of avoiding the predomination by heterotrophs in
standard batch enrichment media containing supplements
such as thiosulfate and glucose or thiosulfate and acetate. In
the latter medium, a mixture of obligately chemolithotrophic
thiobacilli and chemoorganotrophs normally develops.
Acidiphilium acidophilum was originally isolated as a com-
mensal of A. ferrooxidans, although T. organoparus (now
considered to be a strain of A. acidophilum) was enriched
directly from an acid mine water environment. Media suit-
able for the different species are summarized in Kelly and
Wood (1998) and are available from the original literature
describing the species. In addition, culture collection cat-
alogues also provide guidance for media suitable for their
cultures.
Most strains are able to produce colonial growth on
appropriate media solidified with agar. Some strains, espe-
cially those of A. ferrooxidans, grow poorly on agar media.
In some cases, this difficulty is due to the toxicity of agar
hydrolysis products and has been overcome in a number of
ways (e.g., Tuovinen and Kelly, 1973). Toxic effects are gen-
erally avoided by the use of a minimal concentration of agar,
screening of suitable brands of purified agars, use of agarose,
and, in the case of A. ferrooxidans, use of a combination of
media with low agar concentrations, at pH 2.22.5, and with
ferrous sulfate at only about 20 mM. The use of silica gel
media as an alternative is normally unnecessary.
Maintenance procedures
After cultivation on suitable media, most species survive stor-
age at 5 C for periods of weeks to months, especially if the
media for neutrophiles have not become too acidic before
storage. A. ferrooxidans survives in culture on pyrite for very
long periods when stored at 5 to 15 C, and T. plumbophilus
survives for at least a year at room temperature on galena
(Drobner et al., 1992). Many strains have been successfully
lyophilized or have survived storage at 20 C, at liquid nitro-
gen temperature, or in glycerol suspension at 20 C.
Differentiation of the genus Thiobacillus from other
genera
At the time of the previous edition of this Systematics, infor-
mation on ubiquinone and fatty acid content, DNA base
composition, and interspecific DNADNA hybridization had
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3
been obtained for the thiobacilli (Kelly and Harrison, 1989).
This enabled division of the species into distinct groups
on the basis of physiological and biochemical characteris-
tics (Harrison, 1982, 1983; Katayama-Fujimura et al., 1982;
Katayama-Fujimura and Kuraishi, 1983). It has also compli-
cated consolidation of species, as DNADNA hybridization
studies in some cases show considerable diversity among
strains regarded as members of the same species, while also
showing very high levels of similarity between putatively
different species. The confirmation by 16S rRNA sequencing
of the phylogenetic diversity of the formally approved species
now forces major revision of this genus.
Primary separation of most of the former 21 species of
Thiobacillus is relatively easy to achieve by virtue of differ-
ences in gross physiological characteristics, such as pH and
temperature requirements for growth, ability or inability
to grow heterotrophically as well as chemolithotrophically,
and differences in the ability to grow anaerobically with
denitrification or to use elemental sulfur. Most species
designated as Thiobacillus may precipitate elemental sulfur
into the medium during growth on sulfide, thiosulfate,
and, in some cases, trithionate or tetrathionate. One of the
exceptions is T. versutus. Sulfur precipitation is not a highly
distinctive diagnostic characteristic, as it is a variable property
influenced by growth conditions, such as oxygen availability
or perturbation of steady state in chemostat culture. The
precipitation of sulfur is superficially comparable to the
extracellular precipitation observed with species of Chloro-
bium and contrasts with the intracellular accumulation of
sulfur by members of the Chromatiaceae such as Chromatium.
It is likely, however, that some species deposit sulfur inter-
nally, subsequently oxidizing it to sulfate. Thus, T. albertensis
grown on thiosulfate is believed to contain a sulfur granule
bounded by a membrane (Bryant et al., 1983), the appear-
ance of which is correlated with the production of a large
amount of extracellular elemental sulfur at the end of the
growth phase. Morphologically, these inclusions resemble
the sulfur found in Beggiatoa and Thiothrix. These granules
do not appear during mid-log growth (Bryant et al., 1983),
and had not been reported in earlier ultrastructural surveys
of other thiobacilli (Mahoney and Edwards, 1966; Shively
et al., 1970), but may have been observed in T. kabobis
(Reynolds et al., 1981), which is now regarded as a synonym
of T. thiooxidans (Kuenen et al., 1992). The possibility of both
extracellular deposition and intracellular accumulation of
elemental sulfur in some thiobacilli is somewhat of a physio-
logical anomaly, possibly indicative of differing mechanisms
or locations of sulfur compound oxidation. Extracellular
sulfur precipitation is best explained as a consequence of the
 4 Bergeys Manual of Systematics of Archaea and Bacteria

conversion of sulfide or the sulfane groups of thiosulfate (or TABLE 2. Species of Thiobacillus recognized in the first
polythionates) to sulfur at the surface of the cell, presumably edition of this Manual (Kelly and Harrison, 1989), or
described subsequently, showing those species assigned,
in the periplasmic space and catalyzed by enzyme systems or proposed for assignment, to new or different genera
located in or external to the bounding membrane of the cell.
This seems to be more plausible than intracellular genera- Revised genus designations
tion of sulfur and its excretion to the outside (as has indeed Basonym (published or proposed)
been suggested). The physicochemical nature of the sulfur Thiobacillus thioparusa Thiobacillus thioparusT
at the moment of formation is uncertain, and, of course, the
Thiobacillus denitrificansb Thiobacillus denitrificans
cell wall would likely be a barrier to the excretion of large
Thiobacillus aquaesulisc Thiobacillus aquaesulis
granules from inside the cell or even to their generation
in the periplasmic space. The production of intracellular Thiobacillus plumbophilusd Uncertain (currently Thiobacillus)

sulfur from a soluble substrate (thiosulfate) implies intracel- Thiobacillus intermediuse
lular oxidation of that substrate and transport of S0 across Thiobacillus perometabolise
the membrane. Although it is clear that this process would Thiobacillus cuprinuse
involve membrane-associated electron transport systems in Thiobacillus thermosulfatuse
the oxidation of sulfur compounds to sulfate, the mechanism
Thiobacillus delicatusf
remains to be elucidated. Clearly, the present stage of our
Thiobacillus neapolitanusg
knowledge of the mechanism of transport of sulfur and its
compounds and of the biochemistry of their conversion Thiobacillus halophilusg
Thiomonas intermediaT
Thiomonas perometabolis
Thiomonas cuprina
Thiomonas thermosulfata
Thiomonas delicata
Halothiobacillus neapolitanusT
Halothiobacillus halophilus

to sulfate is inadequate. Therefore, characteristics such as Thiobacillus hydrothermalisg Halothiobacillus hydrothermalis

intracellular sulfur accumulation cannot be employed as Thiobacillus tepidariusg Thermithiobacillus tepidariusT
reliable taxonomic features. Thiobacillus thiooxidansg Acidithiobacillus thiooxidansT
Thiobacillus ferrooxidansg Acidithiobacillus ferrooxidans
Taxonomic comments
Thiobacillus caldusg Acidithiobacillus caldus
Since the description of Thiobacillus was published in the Thiobacillus albertensisd (Acidithiobacillus)
first edition of the Systematics (Kelly and Harrison, 1989), the Thiobacillus prosperusd Uncertain (Acidihalobacter)
genus has undergone a number of revisions and emenda- Thiobacillus novellush Starkeya novella
tions. These changes are summarized in Table 1 and Table 2.
Thiobacillus acidophilusi Acidiphilium acidophilum
Table 2 also presents the DNA base composition and class
Thiobacillus versutusj Paracoccus versutus
within the Proteobacteria to which each species is now assigned,
a Kelly and Harrison, 1989.
based on sequence analysis of either partial or complete 16S b Kelly and Wood, 2000a; see text.
rRNA genes and recently proposed reassignments to new c McDonald et al., 1997.
d See text.
genera (Kelly and Wood, 2000a, b; Kelly et al., 2000). A
e Moreira and Amils, 1997.
phylogenetic tree illustrating the new and different genera f Requires investigation.
to which most of the Thiobacillus species have been assigned g Kelly and Wood, 2000b.

is shown in Figure 1. h Kelly et al., 2000.

First, T. versutus and T. acidophilus (both members of
the class Alphaproteobacteria) have been formally transferred
from Thiobacillus into other genera (Katayama et al., 1995;
Hiraishi et al., 1998; see the chapter by Hiraishi and Imhoff
on the genus Acidiphilium in the family Acetobacteraceae in this
volume), and each appears elsewhere within this edition. The
remaining species group within the classes Alphaproteobacteria,
Betaproteobacteria, and Gammaproteobacteria. The type species,
T. thioparus, falls in the Betaproteobacteria, along with the
closely related T. denitrificans and the moderate thermophile
T. aquaesulis (McDonald et al., 1997; Kelly and Wood, 2000a,
b). These three species are the only ones that can be justified
i Hiraishi et al., 1998.
j Katayama et al., 1995.
based on molecular criteria to be members of the genus
Thiobacillus. A fourth validly named species, Thiobacillus
delicatus, is physiologically similar to members of the genus
Thiomonas and may be reassigned to that genus when more
data are available (Y. Katayama, personal communication).
Thiobacillus denitrificans has been regarded as a close relative
of T. thioparus since it was originally isolated by Beijerinck
(1904a, b), and that view is now strongly supported by the
confirmation that it is a member of the Betaproteobacteria

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Bergeys Manual of Systematics of Archaea and Bacteria
FIGURE 1. Phylogenetic tree based on 16S rRNA gene
sequences of representatives of the genera to which species
formerly described as Thiobacillus are now assigned (Thiobacil-
lus, Acidiphilium, Starkeya, Thermithiobacillus, Acidithiobacillus,
Halothiobacillus, and Thiomonas). Reference species are taken
from the Ribosomal Database Project (RDP) or from the
sequence database libraries. T indicates the type strain of
the species. Bar = 10 inferred nucleotide changes per 100
nucleotides. Numbers at some nodes indicate the number of
times the species to the right occurred in 100 bootstrap repli-
cates.
(Lane et al., 1992; H.G. Trper, personal communication,
1999; Kelly and Wood, 2000a). Of the remaining species,
only Thiobacillus novellus has been shown to be a member of
the Alphaproteobacteria. T. novellus is facultatively autotrophic
(as are the Alphaproteobacteria species formerly classified as
T. versutus (now Paracoccus versutus) and T. acidophilus (now
Acidiphilium acidophilum), as well as Thiosphaera pantotropha
(now Paracoccus pantotrophus) and has been proposed to
be the type species of a new genus, Starkeya (Kelly et al.,
2000). Of the other species in the Betaproteobacteria, four
have been reassigned to the new genus Thiomonas (Moreira
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5
and Amils, 1997). A fifth acidophilic, lead-sulfide-leaching
species reportedly also in the Betaproteobacteria, Thiobacillus
plumbophilus (Drobner et al., 1992), has been too little stud-
ied to know if it should also be reassigned to Thiomonas, so for
the present time it is retained in the genus Thiobacillus. Of the
species within the Gammaproteobacteria, a broad division can
be seen between two principal groups: one comprising the
proposed new genus Halothiobacillus (McDonald et al., 1997;
Kelly et al., 1998a; Kelly and Wood, 2000b), and the other
(Acidithiobacillus) containing at least three of the acidophilic
species, some of which also oxidize Fe2+ and sulfide min-
erals (Table 2; Figure 1). The former Thiobacillus tepidarius
also seems to be somewhat distantly related to this group
(McDonald et al., 1997; Goebel et al., 2000), but we consider
its moderate thermophily and lack of acidophily sufficient
grounds to regard it as a distinct genus (Thermithiobacillus)
at this time (Kelly and Wood, 2000b). Thiobacillus albertis
(whose specific epithet is here corrected to albertensis) is
also acidophilic and strictly chemolithotrophic, making it
physiologically similar to Acidithiobacillus thiooxidans, but it
differs from that species in possessing a glycocalyx and having
a mol% G + C of 61.5, which is significantly higher than that
of A. thiooxidans (5052 mol% G + C). Based on 16S rRNA
sequences, we confirm the placement of T. albertensis in
Acidithiobacillus (Table 2), whose members have DNA mol%
G + C values ranging between 5064 (Table 2). T. prosperus
is more difficult to define taxonomically, as it appears to be
completely unrelated to the T. ferrooxidansT. thiooxidansT.
caldus cluster based on 16S rRNA sequence comparisons
(Goebel et al., 2000) and shows negligible DNADNA
hybridization with other thiobacilli. We have reported the
acidophilic species T. ferrooxidans, T. thiooxidans, and T. cal-
dus, as well as T. tepidarius, to fall in the Gammaproteobacteria,
whereas some reports in the literature assign these to the
Betaproteobacteria. It seems clear to us from recent studies that
these species, while close to the betagamma separation, are
rightly placed in the Gammaproteobacteria (McDonald et al.,
1997; Goebel et al., 2000; Kelly and Wood, 2000b).
Descriptions of the former Thiobacillus species T. versutus
and T. acidophilus are found in the chapters on Paracoccus and
Acidiphilium, respectively. Some characteristics of the genus
Thiobacillus of the newly created genera, and of Paracoccus and
Acidiphilium are given in Table 1.
Differentiation of the species of the genus
Thiobacillus
The characteristics of each of the six species are summarized
in Table 3.
 6 Bergeys Manual of Systematics of Archaea and Bacteria

TABLE 3. Basic characteristics of species of the genus Thiobacillus

Characteristic T. thioparus T. aquaesulis T. denitrificans T. plumbophilus T. delicatus T. prosperus

Mol% G + C 6263 66 63 66 6667 64
Cell size (m) 0.5 1.7 0.3 0.9 0.5 1.03.0 0.25 3.0 0.40.6 0.71.6 0.3 34
Motility + + + + +
Carboxysomes + nd nd nd +
Obligately chemolithoautotrophic + + + +
Optimal pH 6.67.2 7.58.0 6.87.4 4.06.5 5.56.0 nd
pH limits 4.57.8 6.59.0 4.58.0 4.06.5 5.07.0 1.04.5
Optimal temperature ( C) 28 4050 2832 2134 3035 37
Nitrate reduction:
To nitrite + + + + nd
To N2 + nd
Growth on:
Hydrogen sulfide + nd + + nd +
Thiosulfate + + + + +
Thiocyanate + + +
PbS nd nd nd + nd +
Methylated sulfides + nd nd nd
Complex media +a nd nd
a Unable to grow on common sugars, organic acids or one-carbon organic substrates.

with growth occurring between pH 4.5 and 7.8. Some strains

List of species of the genus Thiobacillus
Thiobacillus thioparus
Beijerinck 1904b, 597AL
...................................................................................
thi.opar.us. G. n. thios sulfur; M.L. adj. thioparus sulfur
producing.
Rods, averaging 0.5 1.7 m. Motile with a polar flag-
ellum. Gram negative. Colonies grown on thiosulfate agar
(12 mm in diameter) are circular and whitish-yellow due to
precipitated sulfur. Turn pink, then brown on aging, espe-
cially in the center of old colonies. In static culture in liquid
thiosulfate medium, sulfur is precipitated, and the medium
becomes turbid with a pellicle of sulfur and cells. Sulfur
granules and tetrathionate and/or trithionate may accu-
mulate, accompanied by a pH drop to 4.5. In well-aerated
or chemostat culture, sulfur oxidation of thiosulfate to sul-
fate may occur without precipitation. Some strains oxidize
thiocyanate, thiosulfate, trithionate, tetrathionate, sulfur,
and sulfide. Obligately chemolithotrophic and autotrophic.
Ammonium salts and nitrates used as nitrogen sources.
Aerobic. Optimal temperature: 28 C. Optimal pH: 6.67.2,
are claimed to grow at pH 10.0. Found in mud, soil, canal
water, and other freshwater sources. Presumably widely
distributed.
The mol% G + C of the DNA is: 6263 (Bd, Tm ).
Type strain: ATCC 8158, DSM 505.
GenBank accession number (16S rRNA): M79426.
Thiobacillus aquaesulis
Wood and Kelly 1995, 418VP (Effective publication:
Wood and Kelly 1988, 342.)
...................................................................................
a.quae.sulis. L. n. aquae waters; L. n. Sulis pertaining to the
Temple of Sulis Minerva (Minerva, the Roman goddess of wis-
dom); M.L. adj. aquaesulis from the waters of Sulis Minerva.
Short rods, 0.3 0.9 m, containing some polyphos-
phate inclusions. Motile. Gram negative. Non-sporeforming.
Chemolithoautotrophic growth on thiosulfate, trithionate, or
tetrathionate. Colonies on thiosulfate agar at 43 C are small
(12 mm), circular, convex, and smooth, becoming white
or yellow with precipitated sulfur. In liquid batch culture,
sulfur precipitation and a drop in pH without tetrathionate
accumulation occur. Initiates growth at pH 79 (3055 C),

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Bergeys Manual of Systematics of Archaea and Bacteria
dropping the pH to 67. No growth at pH 6.4 or 9.4 or
at 26 C or 58 C. Chemostat cultures do not accumulate
sulfur or other intermediates during growth on thiosulfate,
trithionate, or tetrathionate at pH 7.6 and 43 C. Facultatively
heterotrophic on complex media (yeast extract or nutrient
broth) but unable to grow on common sugars, organic
acids, formate, or methylamine as single substrates. Uses
ammonium salts as nitrogen sources. Capable of anaerobic,
autotrophic growth during thiosulfate-dependent denitrifi-
cation in batch culture, producing nitrite and sulfur from
thiosulfate and nitrate. Optimal temperature: 4050 C.
Optimal pH: 7.58.0. Contains ubiquinone-8. Isolated from
the thermal springs at Bath, Avon, England.
The mol% G + C of the DNA is: 65.7 (Tm ).
Type strain: ATCC 43788, DSM 4255.
GenBank accession number (16S rRNA): U58019.
Thiobacillus delicatus
(ex Mizoguchi, Sato and Okabe 1976)VP
Katayama-Fujimura, Kawashima, Tsuzaki and Kuraishi
1984a, 142.
...................................................................................
del.i.catus. L. masc. adj. delicatus delicate.
Rods, usually single, rarely in pairs, 0.40.6 0.71.6 m.
Nonmotile. Colonies grown on yeast extractthiosulfate
agar (1 mm. diameter) are smooth and circular and change
from transparent to whitish-yellow with sulfur accumulation.
Facultatively chemolithotrophic and mixotrophic. Grow
autotrophically with sulfur, thiosulfate, or tetrathionate, but
not with thiocyanate. Accumulates tetrathionate and trithion-
ate transiently during growth on thiosulfate. Incapable of
heterotrophic growth on single-carbon compounds. Grows
mixotrophically in thiosulfate media supplemented with tri-
carboxylic acid cycle intermediates or amino acids. Optimal
growth requires both organic and inorganic substances and
thiosulfate or sulfur. Facultatively anaerobic; reduces nitrate
and produces nitrite in mixotrophic and autotrophic media
with thiosulfate or tetrathionate. Ammonium salts, nitrate,
urea, glutamate, or aspartate can be used as the nitrogen
source.
Optimal temperature 3035 C, range 1542 C (no
growth at 10 or 45 C). Optimal pH 5.56.0, range 5.07.0.
Isolated from mine water. Distribution unknown.
The mol% G + C of the DNA is: 6667 (Tm , chemical analy-
sis).
Type strain: IAM 12624.
Additional Remarks: Until 16S rDNA sequence data become
available for this species, it cannot be firmly reassigned to the
genus Thiomonas. Physiologically, the species is more similar
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7
to Thiomonas perometabolis than to the type species Thiobacillus
thioparus.
Thiobacillus denitrificans
(ex Beijerinck 1904b) Kelly and Harrison 1989, 1855
emend. Kelly and Wood 2000a, 548VP
...................................................................................
de.ni.trifi.cans. M.L. v. denitrifico denitrify; M.L. part. adj. deni-
trificans denitrifying.
Short rods, 0.5 1.03.0 m. May be motile by means of
a polar flagellum. Under anaerobic conditions, colonies are
clear or weakly opalescent when grown on thiosulfatenitrate
agar, On aging, colonies may become white with sulfur. Vig-
orous nitrogen production when grown under anaerobic
conditions, leading to splitting of agar when solid media
are used. Facultatively anaerobic. Grows autotrophically
and aerobically on thiosulfate or tetrathionate, on which it
produces growth yields approximately double those of T.
thioparus or T. neapolitanus. Grows anaerobically on thio-
sulfate, tetrathionate, or sulfide by using nitrate, nitrite, or
nitrous oxide as the terminal electron acceptor. Oxidizes
sulfur, sulfide, thiosulfate, tetrathionate, and probably sul-
fite, but not thiocyanate. Chemostat culture can be switched
easily and repeatedly between aerobic and anaerobic growth
modes, with adaptation involving derepression of nitrate and
nitrite reductase synthesis. Ammonium salts and, in at least
some strains, nitrate are used as nitrogen sources.
Obligately chemolithotrophic and autotrophic. Optimal
temperature: 2832 C. Optimal pH: 6.87.4. Found in soil,
mud, and freshwater and marine sediments, especially under
anoxic conditions. Probably very widely distributed.
The original isolation of Beijerinck (1904a) may not have
been a pure culture (Vishniac and Santer, 1957), and a viable
sample is not available in the Delft Culture Collection (L.A.
Robertson, personal communication) or in any other culture
collection. Later work has demonstrated unambiguously that
Beijerincks designation was of a legitimate species exhibiting
stable physiological characteristics (Lieske, 1912; Baalsrud
and Baalsrud, 1954; Taylor et al., 1971; Justin and Kelly,
1978; Katayama-Fujimura et al., 1982). Earlier claims that
the capacity of this organism to denitrify was lost on aerobic
subculture and that it was facultatively heterotrophic were
erroneous. The name has therefore been revived by Kelly and
Harrison (1989, in Validation List No. 31), who suggested a
neotype strain. It is now proposed that NCIB 9548 (the strain
isolated by White and Hutchinson and used by Justin and
Kelly, 1978; AB7, ATCC 23644, JCM 3870) be accepted as this
reference strain (Kelly and Wood, 2000a).1
The mol% G + C of the DNA is: 63 (Bd, Tm ).
 8 Bergeys Manual of Systematics of Archaea and Bacteria
Type strain: ATCC 23644, DSM 12475, NCIB 9548.
GenBank accession number (16S rRNA): AJ243144.
Thiobacillus plumbophilus
Drobner, Huber, Rachel and Stetter 1992, 217.
...................................................................................
plum.bophilus. L. neut. n. plumbum lead; Gr. v. philein to love;
M. L. adj. plumbophilus loving lead, referring to its ability to
grow with lead sulfide as a sole energy source.
Cells are rod-shaped, Gram negative, 0.25 3 m, and
motile by one polar flagellum. Optimal growth between
21 C and 34 C, with growth occurring at up to 41 C. Growth
between pH 4.0 and 6.5. Strictly chemolithoautotrophic
and aerobic. Oxidation of galena (PbS), H2 S, and hydro-
gen. Sensitive to ampicillin and rifampicin. Possesses 96.5%
ubiquinone Q-8. Isolated from a uranium mine in Germany.
Insignificant DNA hybridization to A. ferrooxidans and T.
cuprinus.
The mol% G + C of the DNA is: 66 (Tm , HPLC).
Deposited strain: DSM 6690.
Thiobacillus prosperus
Huber and Stetter 1989, 484.
...................................................................................
prospe.rus. L. masc. adj. prosperus prosperous, referring to its
ability to gain precious metals by ore leaching.
Cells are Gram-negative rods, 0.3 34 m. Motile by
one polar flagellum. Optimal growth around 37 C and up to
41 C. Growth from 03.5% NaCl (some strains tolerate 6%
NaCl) and at pH 1.04.5. Strictly chemolithoautotrophic and
aerobic. Growth on sulfidic ores, like pyrite, sphalerite, chal-
copyrite, arsenopyrite, and galena, and on H2 S. Poor growth
on elemental sulfur and ferrous iron. Produces sulfuric acid
from reduced sulfur compounds. Sensitive to ampicillin
and vancomycin. Possesses meso-diaminopimelic acid and
ubiquinone Q-8. Lives in marine sediments in hydrothermal
areas. Insignificant DNA hybridization to T. ferrooxidans, T.
thiooxidans, T. neapolitanus, and T. thioparus. Member of the
class Gammaproteobacteria.
The mol% G + C of the DNA is: 64 (Tm , HPLC).
Deposited strain: DSM 5130.
GenBank accession number (16S rRNA): AY034139.
Additional Remarks: T. prosperus will in due course be
removed from the genus, as recent work has shown its
closest phylogenetic relatives to be Acidihalobacter aerolicus
(DSM 14174) and Acidihalobacter ferrooxidans (DSM 14175),
with both of which it shares 95% 16S rRNA sequence iden-
tity (P.R. Norris, K.B. Hallberg and B. Johnson, personal
communication, 2001).
......................................................................................................................................................................................................
This article is 2005 Bergeys Manual Trust. Published by John Wiley & Sons, Inc., in association with Bergeys Manual Trust.
End note
1. Editorial Note: In the original description, the authors
erroneously designated the neotype strain as NCIMB
8327.
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