Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
_____________________________________________________________________
Algae and Protozoa Communities on Solid Matters in
Jingyuetan Lake, Chuangchun, China
Deming Dong*1, Liang Liu1, Xiuyi Hua1, Fan Yang1, and Shuofei Dong2
1
College of Environment and Resources, Jilin University, 2699 Qianjin Street, Changchun 130012, P. R.
China
2
College of Environment, Beijing University, Beijing 100871, P. R. China
In aquatic environment, microorganisms and their extracellular substances on solid matters as surface
coatings (SC), suspended particulate matters (SPM), and deposited sediments (DS) play important roles in
controlling the transportation and circulation of trace metals in natural waters. In this paper, the microbial
composition of solid matters in Jingyuetan Lake was studied, including identification of algae, protozoa
and metazoa species, and determination of their numbers and preponderant species. In addition, water
qualitiy of Jingyuetan Lake was evaluated based on the microorganisms indication. The results showed
that SC contains abundant algae, protozoa, and metazoa, while both Species and quantity of algae and
protoza on DS were less than SC greatly. As far as SPM, species and quantity of algae and protoza were
in the midist. Evaluation of water quality based on the indication of algae suggested that Jingyuetan Lake
was contaminated with organic pollutants.
Keywords algae; protozoa; SC; SPM; DS
1. Introduction
Solid matters as surface coatings (SC), suspended particulate (SPM) and deposited sediment (DS) were
located in different positions of the same aquatic environment. These solid matters, together with water
phases, constitute the multi-phases system deciding the activities of heavy metals in the aquatic
environment. In aquatic environment, adsorption of heavy metals onto solid matters is very important in
controlling the transportation and circulation of them [1-4]. Owing to the different forming conditions
like temperature, sunlight and hydrodynamic conditions etc, they possessed respective microbial
composition. Microorganisms in the solid matters not only adsorb heavy metals and organic pollutants
directly in aquatic environment, but also influence their behaviors through affecting the forming and
properties of reactive components like Mn oxides, Fe oxides etc in solid matters [5,6]. In addition,
species and quantity of microorganisms, especially algae and protozoa, could reflect the water pollution
well, accordingly they were often used as indicator to study the status of natural water quality. In this
paper, algae and protozoa on the SC, SPM and DS collected from the same water were analyzed
qualitatively and quantitatively.
2. Solid matters in aquatic environment
The SC (biofilms and mineral associated) existed widely on the surface of all kinds of medium in natural
waters. It was generally dominated by vital organic matters like algae and protozoa. It also contains
abiotic organic matters like humus and minerals like Mn oxides and Fe oxides. The oxidation of Mn(II)
on SC is generally kinetically inhibited under circumneutral conditions and requires biological catalysis.
This biological formation mechanism has been shown to strongly influence the resultant adsorption
properties of the Mn oxides [7-9].
The SPM generally suspended in natural waters and 0.45µm micropore membrane is usually employed
to separate SPM in rivers, lakes and oceans [10]. SPM mainly include most minerals, debris of animals
and plants decomposition, bacteria and algae.
*
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©FORMATEX 2007
Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
_____________________________________________________________________
The DS was located at the bottom of natural waters. It generally contained a great deal of minerals and
abiotic organic matters like humus, while content of vital organic matters were relatively less.
3. Identification of algae and protozoa species on solid matters in
Jingyuetan Lake
Here Jingyuetan Lake was chosen as natural waters for collecting SC, SPM and DS in the summer of
2006 to investigate the characteristics of algae and protozoa on solid matters in nature waters. It was
located in the national forest park in the suburb of Changchun City with an area of 4.3 km2.
3.1
Collection of SC, SPM and DS samples
Collection of SC: Precleaned glass microscope slides (5.0×7.5×0.1 cm) were fixed on polypropylene
racks (Chinese patent ZL 01 2 02517.8) and submerged in the water at a depth of approximately 30 cm
for a period of 2 weeks. After retrieval from the lake, SC on the glass slides were scraped off and
transferred into the beaker filled with MMS (Minimal Mineral Salts) solution [11], and then stirred
homogeneously for use.
Collection of SPM: Filter (0.45µm) was used to separate SPM from water, which had been kept still
for a period prior to filtration. MMS solution was used to dilute obtained SPM samples.
The DS samples were collected at a depth from 0 to 2cm, and MMS solution was still used to dilute
obtained DS samples.
Fig. 1 Micrograph of SC, SPM and DS (a —SC，b, c—SPM，d—DS)
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Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
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3.2
Species of algae and protozoa on solid matters in Jingyuetan Lake
Light microscope (COIC，XSJ-2) was used to observe the algae and protozoa on SC, SPM and DS
(16×25), glass slides with SC attached were directly observed under microscope, while for SPM and DS,
suspension of SPM and DS was droped onto the slides and then observed. The algae, protozoa and
metazoan species on three solid matters were identified to genera [12-14]. The results indicated that
species of algae and protozoa included Bacillariophyta division, Chlorophyta division, Cyanophyta
division, Ciliated Protozoa division and Sarcodina division etc. Fig.1 (a, b, c, d) was the micrograph of
solid matters obtained using microcamera and algae and protozoa identified on the SC, SPM and DS
were listed in table1～3. From Fig.1, both appearance and microorganisms species on solid matters were
different. SC contains large amounts of vital organic matters such as algae, protozoa, metazoan, which
could constitute complex minitype ecosystem. Though abundant algae were observed on SPM as well,
the species and quantity of protoza and the metazoan were still less than SC greatly. As far as DS, it was
consisted of minerals particles and organic crumb and only less algae had been found on it.
Tab. 1(a) Identification of algae genera on SC
Division
Bacillariophyta
Chlorophyta
Cyanophyta
Genera
Stephanodiscus Ehr.
Tabellaria Ehr.
Fragilaria Lyngby.
Synedra Ehr.
Achnanthes Bory.
Chlamudomonas Ehr.
Chlorella Beij.
Scenedesmus Mey.
Volvox Linn．
Closterium Nitzsch．
Oscillatoria Vauch．
Gloeocapsa Kütz.
Aphanizomenon Morr.
Stayroneis Ehr.
Navicula Bory.
Pinnularia Ehr.
Cymbella Ag.
Chlorococcum Fries.
Ankistrodesmus Cord．
Pediastrum Mey.
Ulothrix Kütz.
Cosmarium Cord．
Microcystis Kütz.
Merismopedia Mey.
Tab. 1 (b) Identification of protozoa genera on SC
Division
Ciliated Protozoa
Mastigophora
Sarcodina
Genera
Spathidium Dujardin
Litonotus
Paramecium Hill
Zoothamnium Bory
Spirostomum Ehrenberg
Euglena viridis
Peridinium Her.
Genus Amoeba
Leptopharynx Mermod
Chilodonella Strand
Vorticella
Epistylis Ehrenberg
Oikomonas
Genus Raphidiophrys
Tab.1 (c) Identification of metazoan on SC
Division
Rotifera
Genera
Diplois daviesiae
Rotaria citrina
Monostyla furcata
Tab.2 (a) Identification of algae genera on SPM
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Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
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Division
Bacillariophyta
Chlorophyta
Cyanophyta
Euglenophyta
Genera
Tabellaria Ehr.
Fragilaria Lyngby.
Synedra Ehr.
Chlamudomonas Ehr.
Chlorella Beij.
Scenedesmus Mey.
Closterium Nitzsch．
Cosmarium Cord．
Oscillatoria Vauch．
Merismopedia Mey.
Euglena Her.
Stayroneis Ehr.
Cymbella Ag.
Chlorococcum Fries.
Ankistrodesmus Cord．
Pediastrum Mey.
Ulothrix Kütz.
Crucigenia Morr.
Eucapsis Clem.
Tab.2 (b) Identification of protozoa genera on SPM
Division
Mastigophora
Sarcodina
Genera
Euglena viridis
Bodo Ehrenberg
GenusRaphidiophrys
Gymnodinium Stein
Tab. 3 Identification of algae genera on DS
Division
Bacillariophyta
Chlorophyta
Genera
Tabellaria Ehr.
Fragilaria Lyngby
Ulothrix Kütz
Navicula Bory
Synedra Ehr.
According to table1～2, both SC and SPM contain abundant algae and majority of species were the
same, since the solid matters were collected from the same water. Only several algae were different with
each other. For example, Stephanodiscus Ehr., Cymbella Ag., Navicula Bory and Pinnularia Ehr. were
identified on SC, but not found on the SPM, while Crucigenia Morr., Eucapsis Clem and Euglena Her
identified in the SPM were not found in the SC. Compared with SC and SPM, algae on the DS were only
included Bacillariophyta division and Chlorophyta division and the species of algae were less, while
protozoa, metazoa was not found on it. Algae were kinds of phototrophy microorganisms and they could
not survive without sunlight. SC was generally developed on surface of solid mediums in shallow
aquatic environment with enough sunlight and adequate oxygen. However, DS was collected at bottom
of waters with deficient oxygen and less sunlight. So sunlight may be head limiting factor and cause
great difference in microorganisms’ abundance between SC and DS.
3.3
Microorganisms indicator of water quality
The community frame and abundance will be replaced after being polluted. So Algae and protozoa
community and dominant species and genera could act as the indicator microorganisms to the water
pollution. Kolkwitz and Marson primarily presented the viewpoint of contaminative ecosystem and
enumerate different indicator microorganisms in 1908 [15]. In 1969, Palmer listed the most important 60
algae genera resistant to the organic pollution on the basis of 295 research reports [16]. Compared with
the species listed in his research, all algae species determined in this research were included in those
most important 60 genera; moreover, species like Euglenophyta, Chlorella Beij. Navicula Bory, Synedra
Ehr., Scenedesmus Mey and Ankistrodesmus Cord were in the top ten genera. Then it could be deduced
that Jingyuetan Lake was contaminated with organic pollutants.
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Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
_____________________________________________________________________
In this research, the data of BOD5, COD and surfactants of Jingyuetan Lake were measured in the same
period. Compared with Chinese Surface Water Quality Standards (GB3838－2002), Jingyuetan Lake
was contaminated with obvious organic pollutants, which was in accordance with the conclusion
obtained from indicator microorganisms above.
4 Determination of algae and protozoa numbers and dominant species
Haemocytometer (16 medium pane×25 small pane) was used here to count the algae and protozoa on the
solid matters quantitatively. After cover clip was covered on the counting chamber (0.1 mm3) in the
middle of haemacytometer, and a spot of suspension of solid matters was dropped along the edge of
counting chamber. After standing for a while, the algae and protozoa were watched and accounted under
microscope. In this research, algae on solid matters were repeatedly measured for five times and
protozoa were repeatedly measured for ten times. The amount of algae and protozoa per unit quantity of
solid matters (Ni) was calculated as following equation:
Ni =
A × 1000 × V1
V0 × m
(1)
In the equation：
Ni－amount of microorganisms per unit quality of solid matters (total microorganisms, alga, protozoa or
individual species) (cells/g)
A－total amount of microorganisms in the counting chamber (total microorganisms, algae, protozoa or
individual species) (cells)
V1－cubage of volumetric flask (30 ml)
V0－cubage of counting chamber (0.1 mm3)
m －quantity of solid phase sample(g).
Amount of algae on the SC, SPM and DS and the preponderant species were listed in the table 4, and the
amount of protozoa and preponderant species on SC were listed in table 5. Species and quantity of
protozoa on SPM and DS were relatively small and would not be discussed here.
Tab. 4 Amount of algae and dominant species on solid matters
Solid
matters
SC
Total amount of
algae (cells/g)
Bacillariophyta
division (%)
Chlorophyta
division ( %)
Other
divisions(%)
6.0×108
57.1
24.2
18.6
8
35.7
50.9
13.4
75
25
0
SPM
8.0×10
DS
2.0×106
Dominant species and
percentage accounted
Fragilaria Lyngby 38.7 %
Synedra Ehr. 25.7%
Ankistrodesmus Cord 32.6%
Fragilaria Lyngby 50%
Ulothrix Kütz 25 %
Tab. 5 Amount of of protozoa and dominant species on SC
Solid
matters
SC
Total amount of
protozoa (cells/g)
Ciliated Protozoa
(cells/g)
3×107
1.9×107
8
Dominant species and
percentage accounted
Vorticella 47.6 %
Amount of algae on SC was about 6×10 cells/g with a large percentage belonging to Bacillariophyta
division and the preponderant genera on the SC were Fragilaria Lyngby in the Bacillariophyta division.
For SPM, amount of algae was a little higher than that on the SC, but percentage of Chlorophyta division
was the highest and following with Bacillariophyta division. Preponderant genera on the SPM were
Ankistrodesmus Cord in Chlorophyta division (Accounting for 32.6% of total algae on SPM) and
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Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
_____________________________________________________________________
Synedra Ehr. in Bacillariophyta division (Accounting for 25.7% of total algae on SPM). As far as DS,
amount of algae on it was only 2×106cells/g, which was about 2 orders of magnitude lower than the other
solid matters. A preponderant genus on the DS was Ulothrix Kütz in Chlorophyta division (25%) and
Fragilaria Lyngby in Bacillariophyta division (50%). Amount of protozoa on SC was about 3×107cells/g
with a large percentage of Ciliated Protozoa and the preponderant genera on the SC were Vorticella in
Ciliated Protozoa division.
Above all, Bacillariophyta division was the most dominant population for both SC and DS, While for
SPM, Chlorophyta division was the most important population. Hutchinson and Wetzel have
summarized the dominant population in different lakes. For eutrophic lake，Bacillariophyta division
was the most dominant population perennially，especially Synedra Ehr. and Stephanodiscus Ehr. This
community characteristic was very close to the characteristics of algae in Jingyuetan Lake and
accordingly it could also be deduced that Jingyuetan Lake was in the eutrophic status.
5 Comparison of algae and protozoa community on SC between Nanhu
Lake and Jingyuetan Lake
In the actual bio-monitoring，natural and artificial substance including rocks, dyke，docks and slides
were often chosen to collect algae and other microorganisms for analysis. As inserted algae (periphyton)
was often fixed and in the relatively steady status, it could reflect the change of water quality well rather
than plankton. So in this study, SC contains most abundant algae and protozoa and more suitable for
studying the water quality conditions compared with the other solid matters. In our previous study,
characteristics of algae and protozoa community on SC in Nanhu Lake have also been undertaken. In this
paper, comparison of two lakes about microbial composition on SC will be made in the following.
Nanhu Lake was a eutrophic lake located in the south of Changchun City, P.R. China. The lake area is
0.91 km2 and the largest storage capacity is 3106 m3. The average depth is approximately 3 m.
Comparison of microbial composition on SC collected from Nanhu Lake and Jingyuetan Lake showed
microorganism community on SC in two lakes were generally the same. In other words, algae were
mainly involved in Bacillariophyta and Chlorophyta division and protozoa was mainly involved in
Ciliated Protozoa division, which was the same for two lakes. But some genera of algae and protozoa
were different. For example, Fragilaria Lyngby, Synedra Ehr., Cymbella Ag. and Navicula Bory. were
identified in both lakes, while Asterionella, Cocconeis, Gomphonema, Meridion and Cyclotella
determined in Nanhu Lake were not found in Jingyuetan Lake and Stephanodiscus Ehr., Stayroneis Ehr.,
Tabellaria Ehr. and Pinnularia Ehr. present in Jingyuetan Lake were not found in Nanhu Lake.
Similarly, for the Chlorophyta division, Ankistrodesmus Cord, Pediastrum Mey., Chlorella Beij, Ulothrix
Kütz, Scenedesmus Mey and Volvox Lin. were identified on SC in both lakes, whereas Dictyosphaerium,
Oocystis and Stigeoclonium etc. determined in Nanhu Lake were not found in Jingyuetan Lake and
Closterium Nitzsch, Chlorococcum Fries, Cosmarium Cord present in Jingyuetan Lake were not found in
Nanhu Lake. As far as protozoa, Chilodonella Strand, Vorticella, Epistylis Ehrenberg, Litonotus,
Paramecium Hill and Zoothamnium Bory etc. were present on the SC in both lakes, but Halteria,
Cyclidium, Strobilidium, Hemiophrys were determined only in the Nanhu Lake and Spathidium
Dujardin, Spirostomum Ehrenberg, Tetramitus Perty, Leptopharynx Mermod etc. were only detected in
Jingyuetan Lake.
The dominant algae and protozoa species on SC in Nanhu Lake was compared with Jingyuetan Lake
and the results suggested that the most dominant algae species were Chlorella and Ulothrix (Accounting
for 59.3% of the total cell densities of algae) on SC in Nanhu Lake and Fragilaria Lyngby (accounting
for38.7% of the total cell densities of alga) in Jingyuetan Lake. The dominant protoza species were the
same for two lakes, viz. Vorticella.
Above all, microorganisms communities on SC in Jingyuetan Lakes and Nanhu Lake were similar,
which may be resulted from similar climate conditions, environmental background values and trophic
state, viz. eutrophic status. However, some microorganisms genera were different for two lakes due to
different water quality condictions and hydrodynamic conditions etc.
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Communicating Current Research and Educational Topics and Trends in Applied Microbiology
A. Méndez-Vilas (Ed.)
_____________________________________________________________________
Conclusion
Microbial composition of SC, SPM and DS was very different due to the distinct forming conditions like
temperature, sunlight and hydrodynamic conditions. Evaluation of water quality based on the indicator
microorganisms suggested that Jingyuetan Lake was contaminated with organic pollutant.
Microorganisms communities on SC in two lakes were generally the same.
Acknowledgments This research was supported by National Natural Science Foundation (20477014).
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