Environmental Engineering Reference
In-Depth Information
Table 1
Biochemical characteristics of
Bacillus
strains
isolated from oil refi nery waste
Bacilliaceae
. Various studies have been conducted
on the genetic diversity of the genus
Bacillus
.
Using 16S rDNA sequence analysis, Ash et al.
(
1991
) distinguished fi ve phylogenetic groups in
this genus. In turn, Nielsen et al. (
1994
) described
a sixth belonging to the alkaliphilic bacilli.
Then, Xu and Côté (
2003
), using both 16S rDNA
and 16S-23S ITS nucleotide sequences, ana-
lysed the phylogenetic relationship between
bacilli species and divided them into ten distinct
clusters.
Since the genus
Bacillus
includes species that
are of industrial, biotechnological, and environ-
mental interest, as well as clinically important
species, their biochemical characteristics have
been intensively studied. They are widely distrib-
uted in the environment and tolerate various
environmental conditions. The majority is meso-
philes, with an optimal temperature of 30-45 °C;
however, some are thermophiles, growing at
even 65 °C, or psychrophiles, able to grow at
0 °C. They are found growing over a range of pH
from 2 to 11. In general, bacilli are capable of
using simple organic compounds, such as amino
acids, sugars, organic acids, or carbohydrates, as
well as various other substances.
The
Bacillus
strains named T-1, T
Strains
T-1
Test
T
′
-1 I
′
-1a
Gram-staining
+
+
+
Growth temperature:
30 °C
+
+
+
37 °C
+
+
+
45 °C
+
+
+
65 °C
−
−
−
Sporulation
+
+
+
Salinity (% NaCl):
2
+
+
+
4
+
+
+
8
+
+
+
12
+
−
+
Utilization of sodium acetate
+
+
+
Tween 20
+
+
+
Tween 80
+
+
+
Indol production
+
+
+
Glucose (acidifi cation)
+
+
+
Arginine (arginine dihydrolase)
−
−
−
Urea (urease)
−
−
−
Esculin (hydrolysis
ʲ
-glucosidase)
+
+
+
Gelatine (hydrolysis)
+
+
+
Glucose (assimilation)
+
+
+
Arabinose (assimilation)
+
+
+
Mannowe (assimilation)
+
+
+
′
-1, and
Mannitol (assimilation)
+
+
+
I
-1a were isolated from 100-year-old oil refi nery
sludge in Czechowice-Dziedzice (Poland) as
described by Berry et al. (
2006
). The morpho-
logical and biochemical properties of these
strains are listed in Table
1
.
′
N-acetyl-glucosamine (assimilation)
+
+
+
Maltose (assimilation)
+
+
+
Gluconate (assimilation)
+
+
+
Caprate (assimilation)
−
−
−
Adipate (assimilation)
−
+
−
Malaje (assimilation)
+
+
+
Citrate (assimilation)
+
+
+
3
Identifi cation of
Bacillus
Strains
Alcaline phosphatase
+
+
+
Esterase (C4)
+
+
+
Esterase lipase (C8)
+
+
+
Taxonomic analysis of species of genus
Bacillus
carries some diffi culties (Guinebretiere et al.
2001
; De Paolis and Lippi
2008
). They constitute
closely related taxa, and their differentiation
and identifi cation using one method is often
impossible and requires the combination of
biochemical, metabolic, and molecular methods
(Ash et al.
1993
; Chun and Bae
2000
;
Guinebretiere et al.
2001
; Suihko et al.
2004
;
Hutsebaut et al.
2006
; De Paolis and Lippi
2008
;
Płaza et al.
2012
). In our study, three methods
Lipase (C14)
−
−
+
Acid phosphatase
+
+
+
According to Płaza et al. (
2010
)
+ positive reaction; − negative reaction
were used to identify
Bacillus
strains: 16S rDNA
sequencing, the Biolog system, and fatty acid
analysis.
Only one isolate (I
-1a) was successfully
identifi ed as
B. subtilis
by the 16S rDNA gene
sequence analysis, while T-1 and T
′
′
-1 were
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