Genus: Bacillus. Streptomyces. Pseudomonas. Azotobacter.

Structural
characteristi
cs:
Gram-positive.
1
Rod/cocci-shaped.
1
Size: !m in "idth to #
!m in len$th.

Gram-positive.
1
%on-motile.
1
&orm round chal'y
colonies.
(
)ontains branchin$
*lamentous hyphae.
1
Gram-ne$ative.
1
Rod-shaped.
1
)ontains motile polar
+a$ella.
1
Size: ,.- to ,.. !m to
1.- to /., !m.
10
Gram-ne$ative.
1
Rod/cocci-shaped.
1
1ostly contains no +a$ella
2non-motile3.
1
Size: -1, !m.
1
Physiologica
l
characteristi
cs:
)an be obli$ate aerobe
or 4acultative anaerobe.
)ontains supero5ide
dismutase 2S6783 but
only some contains
catalase 2)at83.
1
&orm endospores.
1
)an under$o
4ermentation: produces
$lycerol and
butanediol.

9ner$y source:
)hemical compounds
such as amino acids.
/
)arbon source: or$anic
compounds such as
$lucose.
0
6bli$ate aerobe:
)ontains both
supero5ide
dismutase 2S6783
and catalase 2)at83
enzymes.
1
%on-photosynthetic.
1
Anti-biotic resistant.
1,
9ner$y source:
6r$anic compounds
such as $lucose.
1
)arbon source:
6r$anic compounds:
typically soil-derived
carbohydrates.
1
;ypically obli$ate
aerobe: )ontains both
supero5ide dismutase
2S6783 and catalase
2)at83 enzymes.
1
%on-photosynthetic.
1
Produces acid 4rom
$lucose.
1
7oes not produce
endospores.
1-
9ner$y source:
6r$anic compounds
such as $lucose.
1(
)arbon source:
<nor$anic salts and
or$anic sources such
as citrate and
acetate.
1(
;ypically obli$ate aerobe:
)ontains both supero5ide
dismutase 2S6783 and
catalase 2)at83 enzymes.
1
%on-photosynthetic.
1
7oes not produce
endospores.
1
)an produce cysts.
/
)an under$o nitro$en
*5ation.

9ner$y source: 6r$anic

compounds such as salts
o4 or$anic acids.
/
)arbon source: 6r$anic
compounds such as
su$ars and alcohols.
/
Example of
species.
Bacillus megaterium.
5
Bacillus subtilus.
5
Streptomyces
coelicolor.
13
Streptomyces
antibioticus.
13
Pseudomonas
putida.
19
Pseudomonas
aeruginosa.
16
Pseudomonas
Azotobacter chrooccum.
2
Azotobacter nigricans.
2
!uorescens.
2"
Biochemical Tests:
Bacillus. Streptomyces. Pseudomonas. Azotobacter.
Gram-stainin$.
7ar' purple.
=
7ar' purple.
1
>i$ht pin'.
1.
>i$ht pin'.
1
65idase ;est.
8
#
8
1
8
1.
8
/
)atalase ;est.
8/-
.
8
1
8
1.
8
/
Penicillin ;est.
-
.
8
1
8
1.
-
/
Detailed descriptions of media requirements:
Genus: 7escription o4 media re?uirements:
Bacillus. Bacillus species can $ro" in non-speci*c media: so it can be cultured in nutrient a$ar that is rich in $lucose 24or
carbon and ener$y source3 and nitrate
1
. Althou$h nutrient broth can also be used: it is not ideal because it "ould be
hard 4or Bacillus species to be isolated in the li?uid medium: so the media should be an a$ar plate to ma'e it easier.
7urin$ incubation: the media should be in aerobic condition

. ;his is because althou$h some strains o4 Bacillus are

4acultative anaerobe: they 4unction better in the presence o4 o5y$en and as a result: increases the chance o4
colonies bein$ 4ormed
1
. 1ost Bacillus species are also mesophiles: and can $ro" in the temperature ran$e o4 /,-
0-@): so the media can be incubated at /#@)
=
. &urthermore: they can $ro" in a "ide pA ran$e o4 bet"een -11 so
colonies can still 4orm i4 the nutrient a$ar is neutral 2pA B #.,3
.
.
Streptomyces. Sensitivity a$ar 2S9%S3 is the most appropriate a$ar to culture Streptomyces colonies because the species are
4astidious
1
. Some o4 these speci*c nutrients such as starch and casein are re?uired 4or $ro"th o4 Streptomyces
species: and these components are already in the S9%S media
(
.
Chen incubatin$: the media should be aerobically at -@) and a pA ran$e bet"een ..,-(.,
1
. ;hese are the
optimum conditions re?uired 4or Streptomyces that are 4ound in soil: and because o4 its ecolo$ical habitat: these
ran$es can be very narro".
Pseudomonas. %utrient a$ar is selected because Pseudomonas can $ro" on very minimal nutrients and the species are not
4astidious: so its non-selectivity ma'es it an appropriate media.
1(
Derosine is also added to the nutrient a$ar because
Pseudomonas can brea' the alcohol do"n: and it is ?uite tolerant to alcohol
1-
. As a result: this ma'es it eEective 4or
strains o4 Pseudomonas colonies to be isolated in the media.
;he media should be incubated aerobically at a temperature o4 /,-/@) and the pA should be #., because these
are the optimum conditions that Pseudomonas species can $ro" at.
1(
;his also allo"s us to narro" do"n the
diEerent number o4 colonies $ro"in$ in the media as other bacterial species may not $ro" optimally in these
conditions.
Azotobacter. Azotobacter is 4astidious and re?uires nitro$en in the media to $ro".
1
As a result: the most appropriate media to
culture Azotobacter species is to use Bur'Fs medium because it has nitro$en-containin$ compounds.
1
;o ensure the $ro"th o4 Azotobacter: cultured Bur'Fs medium should be incubated aerobically 2as Azotobacter
species are obli$ate aerobes3
1
: and the temperature should be /,@) 4or colonies to 4orm. ;his is the optimum
temperature that Azotobacter species in the soil metabolise and $ro"

. &urthermore: they have a "ide pA ran$e o4

-.--..- that they can $ro" in: but ideally: Bur'Fs medium should be incubated "ith a pA ran$e o4 #.,-#.- to ensure
optimum conditions 4or the Azotobacter species

.
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