PRODUCTION OF B-VITAMINS BY PLANKTONIC BACTERIA ISOLATED FROM THE MESOTROPHIC LAKE JASNE

Microbiology
PRODUCTION OF B-VITAMINS BY
PLANKTONIC BACTERIA ISOLATED
FROM THE MESOTROPHIC LAKE JASNE
WOJCIECH DONDERSKI*
BARBARA NOWACKA*
SUMMARY : Studies were carried out on the occurrence of organisms producing vitamins of group
B among the heterotrophic planktonic bacteria of the littoral and pelagic zone of the mesotrophic lake
Jasne. In the littoral zone the most numerous group were bacteria producing nicotinic acid, pantothenic
acid and riboflavin. In the pelagic zone the number of B-vitamins producers was much smaller, and the
predominant group were strains synthesizing folic acid, biotin and thiamin. The percentage of B-
vitamins producers in the pelagic zone was higher in spring than in autumn. In the littoral zone the
reverse situation was stated. The majority of planktonic bacteria of both zones were able to produce 2-
4 vitamins. Only a few strains synthesized only one vitamin.
Key words : Planktonic bacteria, heterotrophic bacteria, littoral zone, pelagic zone, B-vitamins.
INTRODUCTION
Heterotrophic bacteria living in water environment
the present work was to detect among the heterotrophic
transform matter and energy cycling in it. This is so,
planktonic bacteria inhabiting various zones and layers of
among other things, because of their abilities, including
water in the mesotrophic lake Jasne strains synthesizing
the ability to synthesize vitamins of group B, contained
some vitamins of group B and to estimate the amounts of
in many enzymes participating in the processes of
vitamins produced by individual strains depending on the
transformation and mineralization of various organic
place and time of their isolation.
compounds (1,13,19, 21).
The presence of vitamins in water environment is one
MATERIALS AND METHODS
of the factors conditioning the growth and development
The subject of the study were heterotrophic bacteria isolated
of many groups of organisms, such as phytoplankton,
from the water of the littoral and pelagic zone of the mesotrophic,
zooplankton, macrophytes and bacteria themselves.
tending towards distrophy, lake Jasne. The lake is a small, forest
According to Ohwada and Taga (20) and Nishijima and
surrounded water basin, 10.7 ha in area, situated in the north-
Hata (17), many algal species need for their develop-
eastern part of Poland, in the Ilawa Lakeland. Its maximum length
is 500 m, its width about 300 m, and its maximum depth 19.8 m.
ment vitamins of group B, separately or in various
The water pH ranges from 4.3 to 4.8. More data about the lake are
combinations. After Hagedorn (11), Gillespie and Morita
given by Zytkowicz (26).
(9) and Kurata et al. (14), quantitative and qualitative
variations in the vitamins in water environment may not
Sampling
only affect growth and productivity of phytoplankton and
Water samples for study were taken at the time of homoth-
macrophytes, but also the seasonal successions of
ermy of the water mass, in spring (April 14) and in autumn
various groups of algae as well.
(October 21) 1990. The samples were taken in the littoral and
The main source of vitamins in water basins is micro-
pelagic zone, from the surface layer (about 30 cm below water
biological synthesis, claim Burkholder (3), Hagedorn (10),
surface), from the medial layer (half way down, at 3.5 m in the
Rheinheimer (22) and Hishijima and Hata (18). The aim of
littoral and 9.0 m in the pelagic zone) and from the near
bottom layer (about 30 cm above sediment - at 7.0 m and 18.0
* From Department of Water Microbiology and Biotechnology,
m deep respectively). The water was collected by means of an
Institute of Biology, Nicolaus Copernicus University, Torun, Poland.
Isatchenko sampler into sterile glass ampoules, which were
Journal of Islamic Academy of Sciences 5:1, 32-38, 1992
32

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BACTERIA ISOLATED FROM THE MESOTROPHIC LAKE JASNE
DONDERSKI, NOWACKA
placed in an ice container (where the temperature was about
Table 1: Number of heterotrophic bacteria synthesizing B-vita-
±7°C), taken to the laboratory and analyzed immediately. The
mins in water of the lake Jasne*.
time between sampling and analyzing did not exceed eight
hours.
Vitamin
Source of
Littoral zone
Pelagic zone
produced
sampling 14.04.90 21.10.90 14.04.90 21.10.90
Isolation and counting of bacteria
a
1.65
19.62
0.17
0.04
For the isolation and counting of bacteria the spread plate
Nicotinic
acid
b
2.33
14.25
0.43
1.40
method was applied. Water samples diluted with sterile buffer
c
2.29
8.99
0.024
0.65
water (4), were plated on the surface iron-peptone agar medium,
after Ferrer, Stapert, Sokolski (8).
a
1.24
15.35
0.07
0.12
Pantothenic
After 10 days of incubation at 20°C the bacterial colonies were
acid
b
1.27
7.98
0.27
1.83
counted, then from each sample 50 colonies were randomly
c
1.31
2.62
0.012
1.29
picked and transferred to a semisolid iron-peptone agar medium
a
0.70
18.76
0.07
0.16
(5.0 g agar/litre). The strains were stored at ±4°C and transferred
Riboflavin
b
1.27
16.52
0.17
1.31
to a fresh semisolid medium every second month to be used for
c
1.31
11.23
0.015
2.21
further studies.
a
0.75
18.76
0.39
1.00
Folic acid
Identification of bacteria
b
1.08
13.66
1.37
9.91
The bacteria were identified according to the scheme
c
1.21
7.12
0.10
12.35
proposed by Shewan, Hobbs and Hodgkins (23), Hendrie, Mitchell
a
0.12
13.63
0.22
0.31
and Shewan (12) and Buchanan and Gibbons (2).
Biotin
b
0.24
6.26
0.67
4.46
Vitamins assay
c
0.05
3.74
0.06
2.48
The methods used to study the synthesis of B-vitamins by the
a
0.16
5.13
0.05
0.26
Thiamine
bacteria under study were the same as described earlier by Strzel-
b
0.13
3.99
0.13
1.42
czyk and Donderski (25).
c
0.05
3.00
0.012
5.77
a
1.98
30.70
0.45
1.24
RESULTS
Total number
In the littoral zone of the lake, in spring the most
of bacteria
b
3.00
23.36
1.50
11.90
numerous group were bacteria producing nicotinic acid
c
2.62
17.97
0.12
17.78
and pantothenic acid (Table 1). In autumn the most
*: number of bacteria x103/cm3 of water a. water from surface layer
numerous were also bacteria synthesizing nicotinic acid,
(about 30 cm depth), b: water middle layer of littoral and pelagic
then riboflavin and folic acid. The least numerous group
zone (respectively: 3,5 and 9 m depth), c: water from the near
both in spring and in autumn were bacteria synthesizing
bottom sediments (about 30 cm above the bottom).
thiamin and biotin but their number in autumn, particularly
of those producing thiamin, was 30-60 times higher than
The quality of vitamins production by the bacteria
in spring in the littoral zone, and 5-480 times in the pelagic
under study was much the same in both lake zones, but
zone. In the vertical plane of the water mass, in the littoral
the quantity varied depending on the place and time of
zone of the lake, in spring the percentage of producers of
isolation of the strains. The bacteria isolated from the
particular vitamins was much alike, while in autumn the
water of the littoral zone, both in spring and in autumn,
numbers of the vitamins producers was generally
produced the largest amounts of pantothenic acid and
decreasing with the depth.
nicotinic acid, and the smallest amounts of thiamin and
In the pelagic zone the number of B-vitamins
biotin (Table 2).
producers among the heterotrophic bacteria was much
The strains isolated from the water of the pelagic zone
smaller. Among bacteria of this zone the predominant
(Table 3) in spring also produced pantothenic acid and
group both in spring and autumn were strains producing
nicotinic acid in largest amounts, and biotin and thiamin in
folic acid. In spring the second largest group were bacte-
smallest amounts, but the amounts of vitamins produced
ria producing biotin and nicotinic acid. In this zone of the
of the bacteria of this zone were 2-14 times smaller than
lake the vitamins producers were generally more numer-
those produced by the strains coming from the littoral
ous in spring than in autumn. The vertical distribution of B-
zone. The bacteria isolated from the water of the pelagic
vitamins producers occurring in the surface layer and
zone in autumn produced pantothenic acid, thiamin and
those above the sediments were but small.
folic acid in the largest amounts. Biotin was also produced
33
Journal of Islamic Academy of Sciences 5:1, 32-38, 1992

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BACTERIA ISOLATED FROM THE MESOTROPHIC LAKE JASNE
DONDERSKI, NOWACKA
Table 2: Amount of vitamins produced by the planktonic bacteria isolated from the littoral zone of the lake Jasne (in ug/g d.w.).
14.041990
21.10.1990
Vitamins
X
Y
Range
Mean
X
Y
Range
Mean
Pantothenic acid
a
11
30.91-7850.00
2720.67
a
7
74.84-4695.49
2225.29
b
11
64.26-7277.42
2130.15
b
6
743.36-5323.31
3245.47
c
8
333.33-11532.35
5131.40
c
6
22.35-13452.63
4892.27
Nicotinic
a
7
142.34-604.84
235.98
a
6
126.37-220.17
183.62
acid
b
7
102.27-6062.50
1246.80
b
9
89.04-564.22
242.01
c
7
98.36-5083.33
1172.79
c
11
38.27-1125.56
381.62
Riboflavin
a
7
26.25-332.93
133.83
a
5
66.96-229.65
124.04
b
5
13.95-206.02
74.58
b
6
26.91-265.31
128.09
c
7
17.82-307.63
97.81
c
9
41.71-205.37
107.31
Folic acid
a
19
12.90-192.10
65.82
a
22
12.00-113.60
59.93
b
18
18.20-145.80
75.17
b
24
11.80-117.10
57.43
c
23
12.00-150.30
72.05
c
19
10.20-177.80
103.80
Thiamine
a
2
6.58-80.63
43.61
a
5
4.18-100.49
29.25
b
1
0.41-0.42
0.41
b
7
4.17-72.53
35.24
c
1
88.82-88.85
88.83
c
6
3.93-46.04
22.49
Biotin
a
3
0.25-1.27
0.85
a
16
0.06-0.84
0.50
b
4
0.26-2.12
0.82
b
11
0.18-0.64
0.35
c
1
0.24-0.26
0.25
c
10
0.01-0.42
0.12
X: source of sampling (as footnote in Table 1), Y: number of strains studied.
but in the smallest amounts. The amounts of thiamin
The most active biotin producers in the littoral zone
produced in autumn by the bacteria of the pelagic
were bacteria of the groups Arthrobacter-Corynebacterium
zone were about 2-14 times as large, and of folic acid
and Flavobacterium-Cytophaga. The largest amounts of
about 3 times as large as those produced by the
folic acid were found in the cultures of Achromobacter and
strains isolated from the littoral zone in the same
Pseudomonas, and nicotinic acid was secreted most
season. The production of biotin by bacteria isolated
intensely by bacteria of the genus Pseudomonas and the
from the pelagic zone was 3-14 times lower than by
group Flavobacterium-Cytophaga. Pantothenic acid was
those from the littoral zone. In the littoral zone more
produced most actively by bacteria of the genera
biotin was produced by bacteria in spring, and less in
Pseudomonas and Alcaligenes, of the group Flavobac-
autumn. In the pelagic zone the reverse phenomenon
terium, Cytophaga and of the family Enterobacteriaceae.
was found.
The most active producers of riboflavin in littoral zone were
It follows from Table 4 that different strains show
bacteria of the genus Pseudomonas and of the group
different abilities to synthesize vitamins of group B. The
Aeromonas-Vibrio, and of thiamin those of the genera
majority of bacteria referred to particular genera or groups
Achromobacter and Pseudomonas.
were able to synthesize 2-4 vitamins simultaneously.
Among bacteria isolated from the water of the pelagic
Strains of the genus Achromobacter, Pseudomonas and
zone most strains produced biotin in similar amounts,
of the group Flavobacterium-Cytophaga in the littoral
but the strains isolated in autumn produced somewhat
zone, as well as those of the family Enterobacteriaceae in
more of it than those isolated in spring. Folic acid was
the pelagic zone were able to produce and secrete
produced most intensely by bacteria of the genus Acine-
simultaneously all six mentioned vitamins. On the other
tobacter, of the group Flavobacterium-Cytophaga and of
hand, bacteria of the genus Bacillus isolated from the
the family Enterobacteriaceae. The most active produc-
water of the littoral zone and those of the genus Alcali-
ers of nicotinic acid proved to be bacteria of the group
genes isolated from the water of the pelagic zone were
Flavobacterium-Cytophaga, the family Enterobacteri-
able to produce only one B-vitamin.
aceae and of the genus Pseudomonas. The largest
Journal of Islamic Academy of Sciences 5:1, 32-38, 1992
34

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BACTERIA ISOLATED FROM THE MESOTROPHIC LAKE JASNE
DONDERSKI, NOWACKA
Table 3: Amount of vitamins produced by the planktonic bacteria isolated from the pelagic zone of the lake Jasne (in ug/g d.w.).
14.04.1990
21.10.1990
Vitamins
X
Y
Range
Mean
X
Y
Range
Mean
Pantothenic acid
a
6
69.24-3592.08
1114.97
a
5
495.65-2427.27
1098.15
b
6
400.00-3975.00
1266.64
b
6
865.38-1243.63
1017.55
c
7
485.52-1640.00
845.36
c
6
51.21-1833.33
1114.44
Nicotinic
a
7
39.21-1309.09
422.90
a
5
25.30-376.47
122.73
acid
b
7
1.54-1650.00
328.72
b
8
26.66-1273.33
353.23
c
8
52.01-800.00
307.72
c
5
148.81-61.53
32.27
Riboflavin
a
7
38.37-1871.55
402.80
a
8
0.74-124.52
38.85
b
5
31.32-150.00
66.07
b
8
24.32-215.87
72.55
c
8
51.02-800.00
307.72
c
5
14.81-61.53
32.27
Folic acid
a
27
18.20-882.40
224.40
a
26
16.90-937.50
189.56
b
25
6.10-708.30
132.50
b
24
19.60-659.40
149.12
c
25
11.40-550.00
186.08
c
25
17.10-860.00
289.48
Thiamine
a
4
5.86-214.29
82.67
a
10
179.14-617.90
407.25
b
3
24.67-211.59
87.74
b
10
38.79-343.75
138.34
c
4
51.56-227.06
169.29
c
8
12.50-124.04
53.86
Biotin
a
7
0.01-0.32
0.09
a
12
0.02-027
0.13
b
8
0.01-0.44
0.06
b
18
0.05-0.51
0.14
c
6
0.01-0.20
0.07
c
6
0.06-0.44
0.23
X: source of sampling (as footnote in Table 1), Y: number of strains studied.
amounts of pantothenic acid were secreted by bacteria of
Kurata, Saraceni and Kadota (15), who, while studying
the family Enterobacteriaceae, of the genus Achromobac-
over the year the content of thiamin, biotin and vitamin
ter and also of the group Flavobacterium-Cytophaga. The
B12 in the horizontal plane of the eutrophic lake Biwa,
most active producers of riboflavin were bacteria of the
found that besides the seasonal variations in their
genus Pseudomonas. Thiamin was produced most
content there were always larger amounts of those
intensely by strains of the groups Aeromonas-Vibrio and
vitamins in the littoral zone than in the pelagic one. This
Flavobacterium-Cytopha-ga and of the genera Acineto-
must explicitly suggest a higher number of producers of
bacter and Achromobacter.
those vitamins in the littoral zone. Among the planktonic
bacteria of the littoral zone (Figure 1) of lake Jasne the
DISCUSSION
predominant strains were those producing nicotinic acid
Many bacterial strains isolated from lakes and seas
and riboflavin. This ability was exhibited by 87.5% and
are able to produce various vitamins of group B (3, 5, 6,
70.7% of bacteria respectively. In the pelagic zone
9,15,16, 24, 25).
(Figure 2) the most numerous group were folic acid
Our studies have demonstrated that in the surface
producers (up to 91.1%), next producers of biotin (up to
water of the littoral zone of lake Jasne bacteria produc-
51.4%), nicotinic acid (37.2%) and thiamin (32.6%). In
ing B-vitamins averaged about 38-49%, and in the
the eutrophic lake Jeziorak, according to Donderski and
water of the pelagic zone about 25-35% of the total
Sokol (6), the producers of nicotinic acid and riboflavin
microflora. In the eutrophic lake Jeziorak, according to
constituted the least numerous group (on average
Donderski and Sokol (6), in the littoral zone there were
about 20% of the total number of bacteria) in the littoral
about 50% of such bacteria, and in the pelagic zone of
zone, while the predominant group consisted of strains
the same lake there were about 40% of them, as
synthesizing biotin and folic acid (40% each on the
reported by Donderski (7). The larger numbers of B-
average). In the pelagic zones of lake Jeziorak (7)
vitamins producers in the littoral zone than in the
similarly as in the lake Jasne, the most numerous group
pelagic zone are in agreement with the data reported by
was that of folic acid producers. This ability was
35
Journal of Islamic Academy of Sciences 5:1, 32-38, 1992

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BACTERIA ISOLATED FROM THE MESOTROPHIC LAKE JASNE
DONDERSKI, NOWACKA
Table 4: Quantity of vitamins produced by different bacteria isolated from the littoral and the pelagic zone of the lake Jasne (in ug/g
d.w.-mean).
Vitamins produced in the littoral zone
Number of
Bacteria
strains
Biotin
Folic acid
Nicotinic acid Pantothen acid
Riboflavin
Thiamine
S
A
S
A
S
A
S
A
S
A
S
A
S
A
Achromobacter sp.
0.368
0.323
74
86
216
337
2716
1851
21
115
88
31
56
65
Acinetobacter sp.
-
-
-
29
-
418
-
-
-
-
-
-
-
3
Aeromonas-Vibrio
-
-
-
67
102
-
1831
-
332
95
-
-
3
2
Alcaligenes sp.
-
-
-
-
355
-
3592
-
-
41
-
-
3
1
Arthrobacter-
1.27
-
58
-
-
-
-
-
-
-
-
-
2
-
Corynebacterium
Bacillus sp.
-
-
-
-
-
-
988
-
-
-
-
-
1
-
Enterobacteriaceae
-
0.412
83
54
369
390
3077
1787
62
109
-
-
22
42
Flavobacterium-
1.19
0.014
56
-
1055
187
3306
-
53
100
-
6
37
4
Cytophaga
Pseudomonas sp.
-
0.156
90
53
1667
154
4342
5567
145
152
40
31
16
74
Vitamins produced in the littoral zone
Achromobacter sp.
0.081
0.206
160
213
234
91
799
1367
66
43
5
273
99
45
Acinetobacter sp.
-
-
-
399
-
43
-
873
-
58
-
398
-
25
Aeromonas-Vibrio
0.045
0.091
8.3
122
-
-
-
495
41
45
-
490
4
7
Alcaligenes sp.
-
-
-
-
-
-
-
-
-
-
115
179
2
1
Arthrobacter-
0.101
-
67
-
-
-
-
-
-
-
-
-
2
-
Corynebacterium
Enterobacteriaceae
0.080
0.151
270
116
484
107
1151
1603
132
43
115
200
70
86
Flavobacterium-
0.013
0.121
213
213
-
661
1282
664
65
131
211
298
13
21
Cytophaga
Pseudomonas sp.
0.132
-
166
69
562
-
764
-
615
175
150
124
22
4
S: spring, A: autumn.
exhibited by about 90% strains in spring and 80% in
synthesized nicotinic acid and riboflavin in largest
autumn. Folic acid producers were also the most numer-
amounts, and biotin, folic acid or pantothenic acid in
ous group in the bottom sediments of both, lake Jeziorak
smallest amounts. The data obtained by Strzelczyk and
(25) and lake Jasne (5).
Donderski (25) for benthic bacteria of the lake Jeziorak
The present studies on the quantities of vitamins of
indicate that they produced nicotinic acid and folic acid
B-group have shown that those varied depending on the
in the largest amounts. On the other hand, the benthic
time and place of sampling and on the genus or group of
bacteria of lake Jasne, according to Donderski and
the bacteria. This corresponds with the results obtained
Strzelczyk (5), secreted the largest amounts of folic
by Donderski and Sokol (6) and Donderski (7) in their
acid and riboflavin. The benthic bacteria of both lakes,
studies on the planktonic bacteria of lake Jeziorak and
according to mentioned authors, produced biotin and
those obtained by Strzelczyk and Donderski (25) and
thiamin in smallest amounts, like in the case of
Donderski and Strzelczyk (5) on benthic bacteria of
planktonic bacteria (Figure 1 and Figure 2).
lakes of different trophies.
It follows from the analysis of the data that amounts
Our studies have demonstrated that in both zones
of vitamins secreted by the planktonic bacteria of the
of the lake Jasne pantothenic acid and nicotinic acid
mesotrophic lake Jasne were in most cases 2-13 times
were secreted in largest amounts, and biotin and
higher than those produced by the benthic bacteria of
thiamin in smallest ones. According to Donderski and
that lake (5), whereas, the amounts of vitamins pro-
Sokol (6), and Donderski (7), the planktonic bacteria of
duced by bacteria isolated from those two environments
the littoral and pelagic zone of lake Jeziorak
in the eutrophic lake Jeziorak were similar (5, 7, 25).
Journal of Islamic Academy of Sciences 5:1, 32-38, 1992
36

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BACTERIA ISOLATED FROM THE MESOTROPHIC LAKE JASNE
DONDERSKI, NOWACKA
Figure 1: Vitamins synthesized by the planktonic bacteria isolated
minative Bacteriology. Williams and Wilkins Comp, Baltimore,
from the littoral zone of the lake Jasne.
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Springfield III, p 133, 1963.
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Bratislava,1967.
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production by heterotrophic benthic bacteria of lakes of
different trophy. II. Mesotrophic lake Jasne. Acta Microbiol
Polon, 4:357, 1987.
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the lake Jeziorak. J Islam Acad Sci, 3:209, 1990.
a: surface water (about 30 cm depth), b: water from the middle layer
7. Donderski W : Production of B-vitamins by heterot-
(about 3.5 m depth), c: water from the near bottom (about 30 cm
above the bottom), N: nicotinic acid, P: pantothenic acid, F: folic
rophic planktonic bacteria isolated from pelagic zone of the
acid, R: riboflavin, T: thiamine, B: biotin.
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Figure 2: Vitamins synthesized by the planktonic bacteria isolated
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Journal of Islamic Academy of Sciences 5:1, 32-38, 1992
38

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Document Outline

• JIAS Volume 5, No 1
• Production of B-Vitamins by Planktonic Bacteria Isolated from The Mesotrophic Lake Jasne
• Summary
• Introduction
• Materials and Methods
• Results
• Discussion
• References

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