Federal Water Pollution Control Administration
Division of Water Quality Research
Analytical Quality Control Laboratory
Cincinnati, Ohio
USE OF A FLOATING PERIPHYTON SAMPLER
FOR WATER POLLUTION SURVEILLANCE
Reprint
February 1970
U.S. DEPARTMENT OF THE INTERIOR
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USE OF A FLOATING PERIPHYTON SAMPLES
FOR WATER POLLUTION SURVEILLANCE
Cornelius I. Weber, Ph.D.
and
Ronald Lo Raschke
Reprint
February 1970
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Preface
This report was first published on September, 1966 as
Applications and Development Report No. 20 of the Water
^
Pollution Surveillance System. The laboratory that operated
the Water Pollution Surveillance System was subsequently
transferred to the Office of Research and Development, and
was renamed the Analytical Quality Control Laboratory.
At the time this report was written, Ronald Raschke was
an aquatic biologist in the plankton laboratory. He later
received his doctorate from Iowa State University, Ames, and
is now on the staff of the Botany Department, Rutgers University,
New Brunswick, New Jersey.
Cornelius I. Weber, Ph.D.
Chief, Biological Methods
Analytical Quality Control
Laboratory
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Table of Contents
Page
1. Introduction 2
2. Methods and Equipment 3
3. Bate of Colonization 5
k. Comparison of Plankton and Periphyton 9
5. Dry and Ash-Free Weights of Periphyton .......... 13
6. Periphyton Diatoms as Pollution Indicators 16
7. Literature Cited 20
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Figures
Page
1. The periphyton sampler • 4
2. Density of live diatoms in periphyton and plankton
samples from the Ohio River at Cincinnati, October
20 - November 21, 19&- 6
3* Rate of colonization of glass slides by pennate diatoms
in the Ohio River at Cincinnati, October 20 - November
21, 19& 7
k. Rate of colonization of glass slides by centric diatoms
in the Ohio River at Cincinnati, October 20 - November
21, 19& 8
5« Dry and ash-free weights of scrapings from glass slides
exposed 2 weeks in the Ohio River at Cincinnati, July 3
to December 10, 1965 I1*-
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Tables
Page
1* Dominant diatom species in perlphyton and plankton
samples from the Ohio River at Cincinnati, October
20 - November 21, 19& 10
2. Live Centric to Live Pennate cell ratios in periphyton
and plankton samples from the Ohio River at Cincinnati 11
3* Percent dead diatom cells in periphyton and plankton
samples from the Ohio River at Cincinnati 12
k. Dry and ash-free weights of periphyton samples from
the Ohio River at Cincinnati 15
5. The most abundant diatom species in periphyton samples
collected in August 1965 above and below a pollution
source in the KLamath River, Oregon ......... 17
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Use of a Floating Periphyton Sampler
for Water Pollution Surveillance
Abstract
A floating sampler was used to collect periphyton in the Ohio
River at Cincinnati. The rate of colonization of glass slides by
diatoms was determined, and the periphyton and plankton diatom
communities were compared. The density of live diatom cells on the
2
slides reached 15,000 per mm in approximately 30 days. The peri-
phyton diatoms were dominated by species of Nitzschia and Navicula,
whereas the plankton diatoms consisted largely of species of
Melosira and Cyclotella. Dry weights of scrapings from slides
n
exposed 1^4- days ranged from 1^9 • 5 mg per slide (32<>5 cm ) in July
1965, to 2.7 mg per slide in December 1965. Ash-free weights
averaged 16.2$ of the dry weight.
Glass slides were exposed in the floating sampler above and
below a group of polluted outfalls on the upper KLamath River,
Oregon. Gomphonema parvulum and Nitzschia palea were the most
abundant diatoms in samples taken in the vicinity of the pollution
sources; whereas Cocconeis placentula was dominant above the out-
falls, and in the oligosaprobic zone downstream.
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Introductiou
Exploratory periphyton studies were Initiated "by the Water
Pollution Surveillance System of the U. S. Public Health Service
in the Fall of 19& to augment phytoplankton data collected from
a nation-vide system of approximately 135 stations*
Man;r aquatic "biologists (FJerdinstad, 196k; Hynes, 19^3;
Novak, 19l*0) have recognized that a satisfactory interpretation
of phytoplankton data obtained for pollution studies from videly
separated river stations is rarely achieved. Attempts to relate
the quality and quantity of algae in grab vater samples to known
or suspected types of pollution are usually confounded by an igno-
rance of the origin and physiological condition of the organisms.
In contrast, the presence of significant quantities of attached
algae on natural or artificial substrates is strong evidence of
the suitability of the vater for the growth of the organisms col-
lected at a station. Inferences regarding vater quality can be
formulated vith greater confidence therefore, if they are based on
the composition and density of the periphyton. Periphyton communities
have long been used by European biologists to characterize pollution
(Kolkvitz and Marsson, 1908; Butcher, 19«*-6, 19^7, 19^9, 1955; FJer-
dinstad, 1950, 1964; siadeckova, 1962; Sladeckova and Sladecek, 1963).
Interest in the periphyton developed later in this country (Blum,
1957; Hohn, 1959; Patrick, 1953, 1957). Except for the vork of Neel
(1953)> the periphyton has received little attention In Federal
pollution studies in the United States.
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Methods and Equipment
The sampler consists of a styrofoam float approximately
12 X 12 X 2 in., supporting a central plexiglass cradle holding
1- X 3-in. glass microscope slides (Figure 1). The construction
materials are commercially available and cost approximately $2.50.
The slides are held with their long axes parallel to, and their
short axes perpendicular to, the water surface.
Slides were removed from the sampler after exposures of 1, k,
7, 15, and 32 days. The periphyton was scraped from the slides
with a razor blade, arid counts were made to determine number of
p
cells per mm of slide area (Weber, 1966). Live and dead diatom
cells were identified to genus on millipore filter preparations
examined at 1000X. Species determinations were made from permanent
(Hyrax) mounts of incinerated diatom materials.
Samples used to determine dry and ash-free weights consisted of
material from four replicate slides, each treated as a separate sub-
sample. These slides were allowed to air dry in the field. The
material was further dried at 50 C in the laboratory and stored in
sealed containers until used. When processed, the slides were
wetted with distilled water, scraped, and the scrapings dried 2k
hours at 105°C and fired for 1 hour at 500°C.
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Figure 1. The periphyton sampler.
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-5-
Rate of Colonization
Following a brief initial lag period, the density of live diatom
cells on the slides increased exponentially, reaching approximately
p
15,000 per mm in 32 days (Figure 2). This rapid rate of increase
indicated that the colonization resulted primarily from the division
of cells which had became attached to the slides during the first few
days of exposure (during the lag period). Diatom counts in the
plankton taken near the sampler remained relatively constant during
the study period. Had the colonization of the slides resulted prin-
cipally from the gradual deposition of drifting cells, a linear rise
in density would have been observed.
Judging from the decline in the growth rate during the latter
part of the exposure period, it was assumed that the population was
largely established within 15 days, and had leveled off at approx-
P
imately 15,000 per mm (32 days). This is similar to the cell density
that Butcher (19^6) found on glass slides exposed 30 days in eutrophic
(oligosaprobic) waters.
It was decided, on the basis of the cell density data, to tenta-
tively adopt a two-week exposure period for the collection of periphyton
samples at Water Pollution Surveillance System stations. This period
would be long enough to permit the development of a populous periphyton
community, yet brief enough to reflect short-term changes in the
water quality.
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D
PERIPHYTON
O TOTAL DIATOMS
PENMATES
CENTRICS
PLANKTON
D TOTAL DIATOMS
_l
z
1C? CO
K?
LU
O
O
id CL
10
15 20 25 30 35
DAYS
Figure 2. Density of live diatoms in periphyton and plankton samples
from the Ohio River at Cincinnati, October 20 - November 21,
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-7-
4
10
OJ.
CO
_J
_)
LU
O
10
,0°
O
A
D
NITZSCHIA
NAVICULA
ACHNANTHES
GOMPHONEMA
SYNEDRA
10 15 20 25 30 35
DAYS
Figure 3. Rate of colonization of glass slides
by pennate diatoms in the Ohio River at-Cincinnati,
October 20 - November 21,
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-8-
CVJ
—^
CO
_J
bJ
O i
10
,0°
MELOSIRA
CYCLOTELLA
5 10 15 20 25 30 35
DAYS
Figure k. Rate of colonization of glass slides
by centric diatoms in the Ohio River at Cincinnati,
October 20 - November 21,
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-9-
The periphyton diatom population was dominated by pennates
during the entire period, They comprised 67$ of the diatoms at 7
days, and approximately 85$ at 15 and 32 days. Gomphonema and
Synedra were the most abundant pennates at 7 days, but the density
of both was later exceeded by Nitzschia, Navicula, and Achnanthes
(Figure 3)« The dominant species were Nitzschia paradoxa Gmel.,
Navicula cryptocephala Kutz., N. tripunctata (0. Mull.) Bory,
Gomphonema parvulum (Kutz.) Grun., G. olivaceum (Lyng.) Kutz.,
Synedra ulna (Nitzsch) Ehr., and Achnanthes sp. The abundance curves
of the centric diatoms are shown in Figure 4. The principal species
were Melosira varians Ag., and Cyclotella meneghiniana Kutz.
Comparison of Plankton and Periphyton
Although colonization of the slides was unquestionably pioneered
by "drifting" organisms, the composition of the periphyton and plankton
diatom communities was very different throughout the entire exposure
period (Table l). In contrast to the periphyton, the plankton diatoms
were principally Gentries, consisting of Melosira ambigua (Grun.) 0.
Mull., M. distans (Ehr.) Kutz., and species of Stephanodlscus and
Cyclotella. This was further evidenced by the ratios of live centric
and pennate cells in the two types of samples shown in Table 2.
The proportions of live and dead diatom cells in the two types
of samples were also of interest (Table 3). Since the organic matter
must be removed from the diatom frustules before species Identifica-
tions can be made, it is not possible to distinguish which cells in
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Table 1. Dominant diatom species in perlphyton and plankton samples from the Ohio River at
1 Day
4 Days
Exposure Period
7 Days
15 Days
32 Days
Periphyton
Synedra
ulna.
Meloslra
granulata
Nitzschla sp.
Navicula sp.
Nitzschla
paradoxa
Melosira
varians
Synedra
ulna.
olivaceum
Gomphonema
parvulum
Synedra
ulna.
Melosira
varians
Nitzschla sp.
Gomphonema
parvulum
Melosira
varians
Navicula
cryptocephala
Nitzschla
paradoxa
Meloslra
varians
Nitzschla
paradoxa
Navicula
tripunctata
Navicula
cryptocephala
Plankton
Meloslra
ambigua
Cyclotella
meneghlnlana
Unknown centric
invisitatus
Stephanodiscus
hantzschli
Meloslra
ambigua
Cyclotella
meneghlnlana
Unknown centric
Melosira
ambigua
Melosira
dlstans
Fragilarla
crotonensis
Cyclotella
meneghlnlana
Melosira
dlstans
Melosira
ambigua
Meloslra
granulata
Unknown centric
Melosira
dlstans
Asterionella
formosa
Melosira
ambigua
Unknown centric
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Table 2, Live Centric to Live Pennate cell ratios in periphyton and plankton
samples from the Ohio River at Cincinnati
Periphyton
Period Exposed
10/20-21/64 (1 day)
10/20-24/64 (4 days)
10/20-27/64 (7 days)
10/20-11/4/64 (15 days)
10/20-11/21/64 (32 days)
Live Live
r!ATvhi*1 f* •T^pnnfl'f^A
1:2.4
1:4.2
1:2.0
1:6.4
1:10.7
Plankton
Date
10/20/64
10/24/64
10/27/64
11/4/64
n/2i/64
Live Live
3.7:1
5.2:1
28.5:1
2.0:1
0.7:1
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Table 3. Percent dead diatom cells in periphyton and plankton
samples from the Ohio River at Cincinnati
Periphyton
Period Exposed
10/20-21/64 (1 day)
10/20-24/64 (4 days)
10/20-27/64 (7 days)
10/20-11/4/64 (15 days)
10/20-11/21/64 (32 days)
% Dead
9
18
2
23
9
mean 12
Plankton
Date
10/20/64
10/24/64
10/27/64
11/4/64
11/21/64
$ Dead
34
35
48
30
12
nean 32
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the permanent mounts vere alive or dead vben the samples were taken.
We have observed that plankton samples usually contain a high per-
centage of dead diatom cells. The mean proportion of dead diatom
cells In plankton samples taken twice Monthly at this station during
the k water years October 1, I960 to September 30, 19& was 29.6)6,
and the proportion of dead diatom cells in the plankton samples
collected during this study averaged 32$. This is a serious weak-
ness in the data, for although it is not likely that the majority
of the cells of the dominant species in a sample would be dead, the
possibility cannot be discounted. Therefore, a high degree of un-
certainty is associated with any interpretation of plankton diatom
species data.
The percentage of dead diatom cells in the periphyton was much
lower, however, averaging only 12$ during the 32-day period dis-
cussed above (Table 3), and 15.6)6 in 12 two-week samples taken in
during the 1965 calendar year. The high percentage of live diatom
cells in the periphyton confers a significantly greater reliability
upon the inferences based on the diatom data from these samples.
Dry and Ash-free Weights of Periphyton
n
Dry weights of scrapings from slides (area, 32.5 cm ) exposed
for two-week intervals from July 3 to December 10, 19&5, decreased
from 1^9.5 mg per slide to 2.8 mg per slide, and ash-free weights
decreased from 21,8 mg per slide to 0.3 mg per slide (Figure 5).
The proportion of organic matter in the samples, however, remained
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8
X
u.
<
UJ
0
rt
\
30
20
5
0 10
0
200
180
160
140
120
100
80
60
40
20
•
" "
• • • • -
•
.
* ^
•
.
i
»
.
j
< o DW
• AFW
•^
1
P
I
•1 i- ' 1
0
N
Figure 5. Dry and ash-free weights of scrapings from glass
slides exposed 2 weeks in the Ohio River at
Cincinnati, July 3 to December 10, 1965.
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Table 4 . Dry and ash-free weights of periphyton samples from the Ohio
River at Cincinnati
Period
Exposed
7/3-7/17/65
7/17-7/31/65
8/14-8/30/65
8/30-9/14/65
10/30-11/14/65
11/26-12/10/65
Dry Wt
(mg),
149.5^3.^
145.6*33-1
73.8tl4.8
48.2120.8
16.5-9.6
2.lto.7
C.V.*
(*)
37.5
22.7
20.0
43.2
56.4
33-3
Ash-Free Wt
(mg)
21.8*6.4
16. 7^3. 6
9.5tl.8
7.ll3.6
4.8t2.7
0. 3^0.2
c.v.
(*)
29.4
21.3
18.9
50.7
56.2
66.7
Ash-Free Wt
Dry Wt
.146
.115
.129
.147
.291
.143
Mean
.162
coefficient of variation.
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relatively constant, averaging 16.2$ (Table *»•). This was consider-
ably lower than the proportion of organic matter found in periphyton
by Newcombe (19^9, 1950) and Nielson (1953), and in the seston of
Wisconsin lakes by Birge and Juday (1922, 193^). The proportion of
organic matter in dried algal cells usually ranges between 50$ and
90$. The value obtained for the periphyton in our study was similar
to that reported by Nelson and Scott (1962) for the seston (12.9$)
in the Oconee River. We have no data on the organic content of the
seston in the Ohio River at Cincinnati. However, the organic con-
tent of the seston in ^9 weekly samples from the nearby Little
Miami River, taken during the 196^-65 water year, averaged 15.8$.
It is likely, therefore, that the major portion of the organic
matter which accumulated on the slides was derived from the seston
(evan though the periphyton was shown to be very different from the
plankton). It was concluded that dry and ash-free weights did not
accurately measure the production of organic matter by periphyton
growing on slides at this station.
Periphyton Diatoms as Pollution Indicators
The utility of periphyton in characterizing pollution is
illustrated by the data obtained from three stations on the upper
reaches of the KLamath River near Klamath Falls, Oregon. These
samples were supplied by personnel from the Klamath Basin Project,
as a part of a cooperative study.
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Table 5. The most abundant diatom species in periphyton samples collected
in August 1965 above and below a pollution source
in the KLamath River, Oregon
Station .1
(7/23-8/6/65)
Species
Cocconeis
placentula
Navicula
Cryptocephala
Nitzschia
oregona
Gomphoneis
herculeana
Others
%
Abun-
dance
28
22
13
9
28
Cells
Per
160
125
Ik
51
502
Station 2
(7/23-8/6/65)
Species
Gomphonema
parvulum
Nitzschia
(palea)
Cocconeis
placentula
Nitzschia
oregona
Others
%
Abun-
dance
44
21
18
5
12
Cells
Per
nm2
2113
1008
864
240
4686
Station 4
(7/9-8/6/65)
Species
Cocconeis
placentula
Nitzschia
oregona
Stephanodis cus
invisitatus
Nitzschia
amphibia
Others
%
Abun-
dance
39
9
6
6
40
Cells
Per
mm,
*
-
-
-
-
*A quantitative comparison of cell densities with this station was
not possible because of the difference in length of the exposure
period.
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The general pattern of water quality at the stations can be
established on the basis of the diatom populations alone, without
other knowledge of environmental conditions (Table 5)» The domi-
nance of Cocconeis placentula (Ehr.) at Station 1, above KLamath
Falls, indicates an abundance of inorganic nutrients and a low
(or moderate) level of dissolved organics. At Station 2, just
below the city, the dominance of Gomphonema parvulum (Kutz.) and
Nitzschia palea (Kutz.) W. Sm. indicates high levels of dissolved
organics (gross organic pollution). The reoccurrence of Cocconeis
placentula as the dominant form at Station ^,30 miles below the
city, is indicative of a return to nearly oligosaprobic conditions,
resulting from oxidation of the organics.
Station 1 is located at the mouth of the Link River, which
drains Upper Klamath Lake, a eutrophic lake with a long history of
nuisance algal blooms. The abundance of Cocconeis placentula at
this station is in agreement with the distribution pattern of this
organism found by Butcher (19^7)> who reported it as a dominant
diatom in oligosaprobic (and eutrophic) waters. Fjerdinstad (1950)
found it in alpha- and beta-mesosaprobic habitats also, which would
explain its occurrence at Station 2, located less than two mi.les
below sewage treatment plant, tallow works, and wood processing
industry waste outfalls. The dominance of Gomphonema parvulum and
Nitzschia palea at Station 2 is a direct result of the effects of
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-19-
the organic pollution. Butcher (19^7) found these two diatoms to
be the most resistant to pollution, and Fjerdinstad (196^) refers
to them as saprophilous, "Occurring most generally in polluted
waters...".
Irrigation return water from the Lost River Basin is discharged
into the Klamath River approximately 11 miles below Station 2.
Another 7 miles downstream the river enters a narrow gorge and
tumbles approximately 2 miles over a rocky bed, falling 300 feet.
Station 4 is located 2 miles below this rapid. Here, Cocconeis
placentula was again the most abundant diatom, accounting for 30$
of the diatom population, whereas Nitzschia palea and Gomphonema
parvulum each comprised only 3$ (included under "others" in Table 5)»
The natural aeration caused by the rapids undoubtedly aided oxidation
of dissolved organics and restoration of oligosaprobic conditions.
The usefulness of the periphyton in determining water quality
is supported by an extensive literature concerning the ecology of
the organisms, which has accumulated during many decades of work by
European aquatic biologists. The examples cited above employed only
the dominant diatom species. A more precise determination of con-
ditions at these stations could have been made by describing the
entire periphyton community.
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Literature Cited
Birge, E. A., and C. Juday. 1922. The inland lakes of Wisconsin.
The Plankton. I. Its quantity and chemical composition. Wis.
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. 1934• Partlculate and dissolved organic matter in inland
lakes. Ecol. Monogr. 4:
Blum, J. L. 1956. The ecology of river algae. Bot. Rev. 22(5):
291-341.
. 1957. An ecological study of the Saline River, Michigan.
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Butcher, R. W. 1946. Studies in the ecology of rivers. VI. The
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. 1947. Studies in the ecology of rivers. VII. The algae of
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. 1955* Relation between the biology and the polluted condition
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FJerdinstad, E. 1950. The microflora of the River MQlleaa, with
special reference to the relation of the benthal algae to pollution.
Folia Limnol. Scand. 5:1-123.
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-21-
FJerdinstad, E. 19&. Pollution of streams estimated by benthal
phytomicro-organisms. I. A saprobic system based on communities
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^9:63-131.
Hohn, M. H. 1959* The use of diatom population as a measure of
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Bynes, H. B. N. 1963. The biology of polluted waters. Liverpool,
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Kolkwltz, R., and M. Marsson. 1908. Okologie der pflanzlichen
Saproblen. Ber. deut. Bot. Ges. 26:505-519.
Neel, J. K. 1953. Certain limnological features of a polluted
irrigation stream. Trans. Amer. Microsco. Soc. 72:119-135.
Nelson, D. J., and D. C. Scott. 1962. Role of detritus in the
productivity of a rock-outcrop community in a piedmont stream.
Limnol. Oceanogr. 7:396-^13.
Newcombe, C. L. 19^9. Attachment materials in relation to water
productivity. Trans. Amer. Microsc. Soc. 68:355-361.
. 1950. A Quantitative study of attachment materials in
Sodon Lake, Michigan. Ecology 31:20*1-215.
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-22-
Nielson, R. S. 1953* Apparatus and methods for the collection of
attachment materials in lakes. Progr. Fish-Cult. 15:87-89.
Novak, W. 19^0. Uber die Verunrelnigung eines kleinen Flusses in
Mahren durch Abwasser von Weissgerbereien, Leder- und Leimfabriken
und anderen Betrieben. Arch. Hydrobiol. 36:386-423.
Patrick, R. 1953* Aquatic organisms as an aid in solving waste
disposal problems. Sew. Ind. Wastes 25:210-217.
. 1957* Diatoms as indicators of changes in environmental
conditions. Syrap. Biological Problems in Water Pollution. U6PH5,
Cincinnati, pp. 71-83.
Sladeckova, A. 1962. Limnological investigation methods for the
periphyton ("Aufwuchs") community. Bot. Rev. 28:286-350.
, and V. Sladecek. 1963* Periphyton as indicator of the
reservoir water quality. I. True-periphyton. Tech. Water
7:507-561.
Weber, C. I. 1966. Methods of Collection and Analysis of Plankton
and Periphyton Samples in the Water Pollution Surveillance System.
Water Pollution Surveillance System Applications and Development
Report No. 19, 19 pp.
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