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SOUTH AFRICAN RIVERINE MACROINVERTEBRATE RESPONSES TO

CHLORINE AND CHLORINATED SEWAGE EFFLUENTS: AN OVERVIEW

Conference Paper · May 2004

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 SOUTH AFRICAN RIVERINE MACROINVERTEBRATE
RESPONSES TO CHLORINE AND CHLORINATED SEWAGE
EFFLUENTS: AN OVERVIEW

M.L. Williams, C.G. Palmer and A.K. Gordon

Unilever Centre for Environmental Water Quality, Institute for Water Research, Rhodes University,
Grahamstown, 6140. E-mail: andrewg@iwr.ru.ac.za

ABSTRACT
Chlorine is widely used in South African sewage treatment works as an oxidising agent and
disinfectant, and the constant input of chlorinated sewage into rivers and streams could potentially
impact riverine ecosystems. The results of acute (96 h) toxicity responses to chlorine by riverine
mayfly nymphs (Baetis harrisoni) collected from both a relatively uncontaminated stream
(Westville) and a more severely impacted river (Umbilo) in KwaZulu-Natal showed nymph LC50
values for free residual chlorine from both sources to be in the region of 4 µg/l. This value
correlates with the South African Water Quality Guidelines’s acute effect value of 5 µg/l for
chlorine. To assess the impact of chlorinated effluent on the riverine ecosystem, the benthic
macroinvertebrate community structure was investigated at several sites up and downstream of
sewage water treatment works on the Umsunduze and Umbilo Rivers, KwaZulu-Natal. Results
suggested that while treated sewage effluent certainly causes changes in macroinvertebrate
community structure, chlorination of this effluent leads to large-scale localised destruction of the
riverine community. This reduces the natural capacity of the riverine community to process organic
waste and recover from the discharge of sewage effluent.

INTRODUCTION
This paper is an overview of a study conducted in the mid 1990’s assessing the responses of
South African freshwater macroinvertebrates to chlorine exposure (1). At the time no data had
been published on this topic, and today, besides the two papers produced as a result of that study
(2; 3) there still appears to be no other published data on the responses of South African
freshwater invertebrates to chlorine. As (3) point out; “to date, [international] chlorine toxicity
research has focused mainly on fouling organisms in industrial cooling water systems (4; 5; 6; 7; 8)
and the effects of paper mill effluent on a number of different organisms (9; 10). Besides
experiments involving Daphnia (11; 12) there appears to have been little [recent] research
investigating the effect of chlorine toxicity on freshwater invertebrates (13; 14; 15), and none on
South African indigenous riverine invertebrates.”

The South African Water Quality Guidelines for Aquatic Ecosystems specify an acute effect value
(AEV) of 5µg/l for chlorine (16). These guidelines were developed to provide a set of water quality
criteria for safeguarding freshwater ecosystems in South Africa. Due to the paucity of data on
responses of indigenous aquatic organisms to chlorine, the Guidelines were developed using
international data, but it was noted that the data used did not satisfy the minimum acute database
requirement (16). It is of interest then to refine the Water Quality Guidelines to reflect actual
tolerances of indigenous organisms within the local environment. In order to investigate this, both
toxicological (3) and ecotoxicological (2) approaches were followed. The toxicological aspect
involved the selection of a macroinvertebrate, the mayfly Baetis harrisoni (Barnard), and the
determination of its acute (96hr) LC50 response value to chlorine. However, wild populations that
are constantly exposed to a pollutant may also become resistant, and for this reason B. harrisoni
nymphs were used from both a relatively uncontaminated stream in Westville and from the more
severely impacted Umbilo River, both in Durban, KwaZulu-Natal (3). The ecotoxicological study
involved an investigation of the changes in macroinvertebrate community structure in response to

Proceedings of the 2004 Water Institute of Southern Africa (WISA) Biennial Conference 2 –6 May 2004
ISBN: 1-920-01728-3 Cape Town, South Africa
Produced by: Document Transformation Technologies Organised by Event Dynamics
322
chlorinated, treated sewage effluent in the Umsunduze and Umbilo Rivers, and also a manipulated
exposure to unchlorinated, treated sewage (2).

METHODS

Toxicology Study
Trials by (1) showed that a recirculating stream was not suitable for testing a volatile chemical such
as chlorine. Consequently, a flow-through artificial stream system was developed and set up at the
Process Evaluation Laboratory at the Wiggins Waterworks in the Cato Manor area near Durban,
KwaZulu-Natal (Figure 1). The experimental system is described in detail in (1) and (3). Briefly,
each stream channel comprised a 1m length of white PVC flat-bottomed guttering. Incoming raw
water from either the Nagle or Inanda reservoirs was drawn off and directed into a 500l fibreglass
sump tank positioned below the stand supporting the artificial streams. Water from the sump was
pumped to each artificial stream and intravenous drip bags were used to provide a steady flow of
sodium hypochlorite using Baxter CONTROL-A-FLOW regulators. The residence time of the
chlorine within the artificial streams was not long enough for the reactions leading to combined
chlorine forms to occur, as a result free residual chlorine (FRC) and total residual chlorine (TRC)
values were the same. Chlorine concentrations were reported as TRC, but it is important to note
that this value reflects only the FRC fraction.

Nagle Reservoir

eR
n du z
Umsu
R

PIETERMARITZBURG
ni
ge

Darville Sewage
M

u
Treatment Works Inanda Reservoir

Westville
Stream

Umbilo Wastewater Cato Manor DURBAN

SOUTH Treatment Works
AFRICA m
U

bil
oR
Study
area

0 20
Kilometres

Figure 1. Map of the study area, KwaZulu-Natal, South Africa.

Mayflies collected from the Umbilo River were exposed to three chlorine concentrations (4, 8 and
12 µg/l). The control was replicated in triplicate and each of the chlorine concentrations had 6
replicates, resulting in 21 artificial streams. Westville Stream mayflies were exposed to five chlorine
concentrations (4, 6, 8, 12 and 16 µg/l). The control and each of the chlorine concentrations were
replicated in triplicate, resulting in 18 artificial streams. Between 35 and 90 nymphs were placed in
each artificial stream, and after a 48hr acclimation period any dead nymphs were removed.
Chlorine release was then effected by dripping in the hypochlorite solutions. Mortality was
monitored after 2hr, 4hr, 6hr, 8hr, 24hr, 48hr and 96hr. Only positively identified B. harrisoni
nymphs were used in mortality computations. LC50 values were determined using the US EPA
Probit Analysis Program Version 1.5 as the data was determined to be parametric (3).

Ecotoxicology Study
To determine the effect of chlorinated sewage effluent on macroinvertebrate communities, two
rivers possessing active sewage treatment works were identified. In the Umsunduze River

323
(Figure 1), macroinvertebrate communities were sampled at two sites upstream of the Darville
Sewage Works outlet (Figure1), and at five sites downstream (see Table 1 for brief notes on the
layout of sites). Sampling was undertaken once during spring 1993 and winter 1994. In the Umbilo
River, sampling occurred on two occasions during winter 1994 from 7 sites, four upstream and
three downstream of the chlorinated sewage outlet from the Umbilo Wastewater Treatment Works
(WWTW) (Figure1) (see Table 2 for brief notes on the layout of sites).

In order to assess the effect of unchlorinated sewage on macroinvertebrate communities the

Umbilo WWTW was granted permission to discharge unchlorinated treated effluent into the Umbilo
River at a site upstream of the chlorinated sewage outlet (the rationale being that any surviving
pathogens would be eliminated by the chlorine 90 m downstream). Although the volume of this
unchlorinated effluent was considerably less than that of the chlorinated effluent (which at times
contributed about half of the flow of the river) it nevertheless was a source of unchlorinated effluent
and it flowed directly into a riffle (sampling Site 3). Sampling of all seven sites was undertaken
twice during spring 1994. A further study assessing the impact of unchlorinated effluent was
conducted during the summer of 1995 and is described and discussed in detail in (2).

The same sampling protocol was followed on all occasions. Three randomly selected samples
were collected from a riffle and 3 from a pool at each sampling site. Macroinvertebrate community
structure was analysed as: a) percent occurrence (17), b) the number of taxa and c) the total
number of specimens found at each site (2).

In the case of the Umbilo River samples, two-way indicator species analysis (TWINSPAN) was
undertaken (18). TWINSPAN arranges sample sites into two distinct groups along the first axis of a
reciprocal averaging ordination. Further dichotomous divisions are achieved using the presence,
absence and relative abundances of indicator species, and groups of species with similar
distribution among sites are produced.

RESULTS

Toxicology Study
Baetis harrisoni from the severely impacted Umbilo River were reported as having a LC50 value of
4.8 µg/l TRC with upper and lower 95 % confidence limits of 5.1 µg/l and 4.5 µg/l. Baetis harrisoni
from the relatively uncontaminated Westville stream were reported having a LC50 value of 4.1 µg/l
TRC with upper and lower 95 % confidence limits of 4.2 µg/l and 3.9 µg/l. When considering the
96hr concentration response curves; that of the Umbilo B. harrisoni fell to the right of B. harrisoni
from the Westville Stream (3).

Ecotoxicology Study

Umsunduze River
At Sites 1 and 2, upstream of the Darville Sewage Works, relatively high FRC and TRC
concentrations were measured (Table 1), possibly as a result of industrial contamination from
further upstream (1). As expected, high chlorine concentrations were measured in the chlorinated
effluent and at sites immediately downstream of the outlet. By Site 7, 7 km downstream of the
outlet, chlorine concentrations had returned to, or were even lower, than sites upstream of the
sewage works (Table 1).

The average number of macroinvertebrate taxa and individuals sampled at each of the sites during
spring and winter generally exhibited similar trends (Figure 2). At sites above the chlorinated
effluent the number of taxa and individuals were greater than at sites immediately below the
outflow. Then with increasing distance from the effluent outflow, the number of taxa and individuals
increased (2). Dominant species above the chlorinated effluent outflow were Orthocladiinae in
spring and Baetis harrisoni in winter (Table 1). Dominant species immediately below the effluent
outflow changed to Chironomus for both seasons. Only in the spring sample did Orthocladiinae
revert to the dominant species further downstream.

324
 Table 1. Determinants recorded at sample sites on the Umsunduze River at time of collection of box samples.
Chlor.e
Sites 1 2 3 4 5 6 7 8
ff.
Location
relative to 2 km U 5mU 5mD 20 m D 2.5 km D 3 km D 7 km D
effluent

Free S 50 50 120 100 500 100 100 0

residual
chlorine
(µg/l) W 20 40 120 100 60 20 20 20 100

Total
S 100 150 1200 900 650 300 40 50

325
residual
chlorine
W 20 80 1100 800 700 100 200 20 90
(µg/l)
Most Orthocladiin- Orthocladiin- Not Orthocladiin-
S All dead Chironomus Not sampled Not sampled
dominant ae ae sampled ae
species Not
sampled W B. harrisoni B. harrisoni Chironomus Chironomus Chironomus Oligochaete Chironomus
sampled
S = Spring 1993 (October)
W = Winter 1994 (June)
U = Sites upstream from chlorinated sewage effluent discharge point
D = Sites downstream from the chlorinated sewage effluent discharge point
Table adapted from (2)
 Table 2. Range of determinants recorded at sample sites in the Umbilo River at the time of collection of box samples.
Un- Chlor. effluent 7
Sites 1 2 chlor 3 4 5 6
Eff. New Old
Location
relative to 250 m U 6mU 1mB 90 m B 5mD 470 m D 6.5 km D
effluent
Free 100- 0-20
W 0-20 0-20 0 0-20 0-20 0-300 20-100 20-80
residual 150
chlorine 100- 40
(µg/l) S 20-70 0-40 0 0-40 20-40 80-25 60-200 70-80
200
Total 100- 0- 40-90
W 0-30 0-20 0 0-60 0-400 90-2500 90-2000
residual 3000 6000
chlorine 900- 800-

326
(µg/l) S 10-270 40-120 0 40-120 60-90 900-1500 30-600 80
2000 2000
B. harrisoni
B. harrisoni B. harrisoni B. harrisoni B. harrisoni Orthocladiin- Orthocladiinae
Ephydrae
W Orthoclad- Orthoclad- Orthoclad- Orthoclad- ae Tanytarsini
Chironomus
Most iinae iinae iinae iinae Chironomini B. harrisoni
Oligochaete
dominant
B. harrisoni B. harrisoni B. harrisoni
B. harrisoni
species Chironomini
Orthoclad- Orthoclad- Orthoclad-
Orthoclad- Chironomini
S Chironomini Orthocladiin-
iinae iinae iinae iinae Orthocladiinae
ae
Chironomi-ni Chironomi-ni Chironomini
Chironomini
W = Winter 1994 D = Sites downstream from the chlorinated sewage effluent
S = Spring 1994 discharge point
U = Sites upstream from unchlorinated sewage effluent discharge point Old = Old sewage plant
B = Sites between the unchlorinated and chlorinated sewage outfalls New = New sewage plant
Table adapted from (2)

326
 Figure 2. Average number of taxa and individuals (± standard deviation) sampled at sites up and
downstream of chlorinated effluent inflow in the Umsunduze River, KwaZulu-Natal.

Umbilo River
Free residual chlorine concentrations at sites above the Umbilo WWTW varied between 0 to 70
µg/l, and TRC between 0 and 120 µg/l. Both FRC and TRC concentrations were significantly
higher at sites immediately downstream of the chlorinated effluent outlet, but had decreased by
Site 7, 6,5 km downstream (Table 2).

Figure 3. Results of a TWINSPAN classification of benthic riffle and pool fauna collected from the Umbilo
River (1994,1995). Source (2).

Variations in the average number of individuals and taxa sampled at the different sites along the
Umbilo River were confounded by chlorine dosing problems and building operations experienced
at the WWTW during this period (1). However, the following general observations were made: Sites
upstream of the chlorinated effluent outflow recorded higher numbers of individuals and taxa
compared with sites immediately downstream. The numbers of individuals and taxa had increased
by Site 7, 6.5 km downstream. The dominant species at sites above the chlorinated effluent were
B. harrisoni, Orthocladiinae and Chironomini. Below the chlorinated effluent Orthocladiinae and

327
Chironomini dominated. Baetis harrisoni was still sampled but in smaller numbers (Table 2).
Regarding the unchlorinated effluent experiment, it was observed that the effluent appeared to
reduce numbers or organisms but did not change the community structure to any great degree (2).

DISCUSSION
The comparative toxicology study of B. harrisoni from the relatively uncontaminated Westville
Stream and the more severely impacted Umbilo River yielded slightly different LC50 values,
suggesting that the Umbilo B. harrisoni were possibly more tolerant of chlorine. The 96hr
concentration response curves, however, suggested this difference was not significant as standard
deviations of cumulative mortalities at specific chlorine concentrations overlapped (3).

The US EPA freshwater criteria for protection of most aquatic species is 11 µg/l TRC as a 96hr
average. In South Africa the acute effect value (AEV) guideline is 5 µg/l TRC (16). The LC50 values
for Umbilo and Westville B. harrisoni were 4.8 µg/l and 4.1 µg/l TRC respectively, indicating that
the AEV guideline of 5 µg/l TRC appeared in this case to be the correct order of magnitude.
However, the occurrence of large populations of B. harrisoni at sites in the Umsunduze and Umbilo
Rivers where the instream TRC and FRC were measured at concentrations an order of magnitude
higher that the Mayflys’ LC50 raises concerns over the sensitivity of organisms undergoing
laboratory toxicity testing (19). The stresses experienced by organisms during collection,
transportation and testing could result in oversensitivity to the contaminants being tested.
Consequently, the South African guidelines should be revised and refined whenever new data
becomes available.

The results of the ecotoxicology study demonstrated clearly the deleterious effect of chlorinated,
treated sewage on riverine macroinvertebrate community structures. In both the Umsunduze and
the Umbilo Rivers, chlorinated sewage effluent often killed entire communities at the point of
discharge, and it was some distance downstream before the communities were restored. In the
Umsunduze River the community structure 7 km from the sewage outlet was still very different
from the communities upstream of the effluent. In the Umbilo River, the communities 6.5 km
downstream from the chlorinated effluent outlet had shown some degree of recovery (2).

The results of the TWINSPAN analysis of samples from the Umbilo River also confirmed the
adverse effects of chlorine in sewage effluent. In the Umbilo River the difference in assemblage
composition between riffle and pools was more distinctive than the effects of effluent discharge and
was responsible for the primary classification split. However, within both riffle and pool samples,
the effects of chlorination were evident. Samples from Site 5 and 6 (just downstream from
chlorinated sewage outflow) had few organisms, low diversity and were dominated by Chironomus.
It is interesting that samples collected from Site 3 and 4 which had been exposed to unchlorinated
effluent were not distinguishable on the basis of faunal composition, indicating the effects of
chlorination as distinct from sewage effluent (2).

REFERENCES
1. M.L. Williams, “Macroinvertebrate Community and Species Responses to Chlorinated Sewage
Effluent in the Umsunduze and Umbilo Rivers, KwaZulu-Natal, South Africa”. MSc Thesis, Rhodes
University, p222 (1996).
2. C.G. Palmer et al., Water SA, 29 p.473 (2003).
3. M.L. Williams et al., Water SA, 29 p.483 (2003).
4. J.S. Mattice & H.E. Zittel, Journal of Water Pollution Control Federation, 48 p.2284 (1976).
5. F.G. Doherty et al., Archives of Environmental Contamination and Toxicology, 15 p.535 (1986).
6. S. Rajagopal et al., Aquatic Toxicology, 39 p.135 (1997).
7. S. Rajagopal et al., Biofouling, 18 p.55 (2002).
8. S. Rajagopal et al., Marine Environmental Research, 55 p.277 (2003).
9. D.P. Middaugh et al., Archives of Environmental Contamination and Toxicology, 32 p.367 (1997).
10. M.R. van den Heuvel et al., Ecotoxicology and Environmental Safety, 51 p.65 (2002).
11. P.A. Taylor, Environmental Toxicology and Chemistry, 12 p.925 (1993).
12. D.J. Fisher et al., Water Research, 33 p.760 (1999).

328
 13. US EPA, “Ambient Water Quality Criteria for Chlorine”, National Technical Information Service, EPA
440/5-84-030 (1984).
14. R.W. Ward and G.M. de Graeve, Water Research Bulletin, 14 p.696 (1978).
15. R.W. Ward and G.M. de Graeve, Water Research Bulletin, 16 p.41 (1980).
16. DWAF, “South African Water Quality Guidelines. Volume 7. Aquatic Ecosystems”, DWAF, Pretoria
(1996).
17. M.J. Swift et al. “Research in Artificial Streams: Applications, Uses and Abuses”, pub Journal of the
North American Benthological Society, (ed G.A. Lamberti) 12 p.313 (1993).
18. M.O. Hill, “TWINSPAN - a Fortran Program for Arranging Multivariate Data in an Ordered Two-way
Table by Classification of the Individuals and Attributes. Unpublished report, Ecology and
Systematics, Cornell University (1979).
19. P.M. Chapman, Environmental Toxicology and Chemistry, 14 p.927 (1995).

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