J Appl Phycol (2013) 25:661–665

DOI 10.1007/s10811-012-9900-7

Influence of barley straw (Hordeum vulgare L.) extract

on phytoplankton dominated by Scenedesmus species
in laboratory conditions: the importance
of the extraction duration
Wojciech Pęczuła

Received: 25 June 2012 / Revised and accepted: 27 August 2012 / Published online: 20 September 2012
# The Author(s) 2012. This article is published with open access at Springerlink.com

Abstract The response of a natural phytoplankton assem- reducing this phenomenon. One promising technique is an
blage dominated by algae of the genus Scenedesmus to the application of barley straw, which has been used in Britain
addition of barley straw extract was studied in a laboratory since the 1980s (Welch et al. 1990). The idea of a cheap,
experiment. The aim of the study was to compare the inhib- simple and safe method of reducing algal blooms caused
iting effect of water extracts obtained by soaking the straw extensive research on its application (for review, see
for 1, 2 and 3 months. We analysed the response of four Ó hUallacháin and Fenton 2010). There have been attempts
species, Scenedesmus subspicatus, Scenedesmus ecornis, to apply barley straw directly in channels (Welch et al. 1990;
Scenedesmus quadricauda and Scenedesmus acuminatus, Caffrey and Monahan 1999), ponds (Boylan and Morris
during 14 days of their exposure to different types of barley 2003; Butler et al. 2005), lakes (Harriman et al. 1997;
straw extract. S. subspicatus and S. ecornis responded Wiśniewski 2002) and drinking water reservoirs (Everall
with decreasing numbers only to the addition of the 3- and Lees 1997; Barett et al. 1999). Laboratory research
month solution (ANOVA; F0290.1, p <0.001; and F011.8, confirmed the inhibitory effects of barley straw on selected
p <0.01, respectively); the two other species were inhibited species of algae (Gibson et al. 1990; Newman and Barrett
by all types of extracts. The results indicate the need for 1993; Martin and Ridge 1999). After several years of re-
more research on the importance of extraction duration to search, it is recognised that the method has limitations and
the inhibitory abilities of barley straw which can be applied drawbacks. The results of some work shows that the use of
in the management of water quality in water bodies. barley straw in some cases has no effect on selected species
of phytoplankton (Boylan and Morris 2003; Brownlee et al.
Keywords Barley straw . Extract . Phytoplankton . 2003; Ferrier et al. 2005) or can even stimulate the algae, for
Scenedesmus example, the bloom-forming cyanobacteria Planktothrix
aghardii and Microcystis aeruginosa (Molversmyr 2002).
Another problem is the dose of straw that should be used in
Introduction a water body. The effective dose that limits algal blooms is
estimated at 250–300 kg of straw ha−1 (McComas 2003),
Although the mass occurrence of planktonic algae in lakes which in turn makes this method neither cheap nor easy to
and reservoirs has been for many years one of the main use in large lakes or reservoirs. Therefore, some laboratory
problems in water quality management, there is no generally studies have focused on the use of aqueous extract of barley
accepted, inexpensive and environmentally safe method of straw, which would facilitate its use in ponds with a smaller
area. So far, the main approaches in the research seem to be
W. Pęczuła (*) the impact of the extract on various species of algae (Ball et
Department of Hydrobiology, University of Life Sciences,
al. 2001; Brownlee et al. 2003; Terlizzi et al. 2002; Ferrier et
ul. Dobrzańskiego 37,
20-062 Lublin, Poland al. 2005) or the chemical composition of the extract (Ridge
e-mail: [email protected] and Pillinger 1996; Waybright et al. 2009; Murray et al.
662 J Appl Phycol (2013) 25:661–665

2010). An important practical issue required for the proper The influence of the extract on the phytoplankton assem-
use of straw is the time needed to obtain an inhibitory effect blage was tested in 24 PET bottles with a capacity of 1.5 L.
(Ó hUallacháin and Fenton 2010). Observations on the The same amount of algal culture (0.4 L) was poured into 18
barley straw application in field studies have shown that bottles, as well as 0.2 L of different types of extracts in
the washout time of inhibiting substances is usually several duplicate. This way, each type of extract (1m, 2m and 3m)
months (Welch et al.1990; Everall and Lees 1997; Harriman was tested in a total of six experimental replicates. The dose
et al. 1997.). However, the problem of time needed to of the extract was calculated to correspond to 1,660 g of
achieve an effect on algae is rarely considered in laboratory barley straw added to 1 L of water. Six bottles served as
studies. Gibson et al. (1990) studied the effect of the straw controls and were filled with 0.4 L of enriched seston and
decomposition degree on its suppressing ability, stating that 0.2 L of dechlorinated water. The experiment was conducted
6-month incubation is most effective on filamentous algae. in September 2010 and lasted 14 days. Bottles were kept
There are no similar studies concerning barley straw extracts under the same conditions as the previous aquaria of algae
and their effects on phytoplankton, especially those taking and were shaken every day. Temperatures during the exper-
into consideration short-time extraction. The aim of this iment ranged from 20.7 to 21.6 °C.
research was to compare the anti-algal activity of the extract Initially (in the control suspension) and at the end (in all
obtained by soaking the barley straw for varying lengths of bottles) of the experiment, pH and conductivity using the
time (1–3 months) and to test the hypothesis that a short- YSI 556 Multi Probe (MPS, USA) and total organic carbon
time extraction can also produce a liquid with algae- concentration using the UV analyser (Pastel, France) were
suppressing ability. The effect of the extract was tested on measured. Species composition of phytoplankton in 100-mL
a natural phytoplankton assemblage composed mainly of samples taken at the beginning and at the end of the exper-
green algae of the genus Scenedesmus in laboratory exper- iment was based on microscopic analysis using an inverted
iment conditions. microscope and the Ütermohl method (Vollenweider 1969).
Abundances are expressed as colonies (coenobia) mL−1. In
order to determine the significance of differences between
Materials and methods the control and experimental assemblages, ANOVA analysis
with Tukey's test was performed using Statistica 6.0 soft-
For the extraction, we have used barley straw (Hordeum ware (Statsoft Inc., USA).
vulgare L.) obtained from a 2009 crop and kept under dry
conditions for a period of 10 months after harvesting. We
prepared three sets of aquaria consisting of three tanks, each Results
with a capacity of 16 L. The same quantity of 80 g of straw
was placed in a set of three at monthly intervals, so the The phytoplankton community after 14 days in the labora-
resulting dose of straw amounted to 5 g of straw L−1 in each tory consisted mainly of green algae from the genus
aquarium. The tanks were filled with dechlorinated tap Scendesmus as well as small numbers of Coelastrum and
water and then continuously aerated using a standard aquar- Ankistrodemsus species. The genus Scendesmus was repre-
ium aerator and kept at room temperature (~20–22 °C) for 1, sented by four species: Scendesmus subspicatus Chodat
2 or 3 months. After the extraction ended, 0.2 L of each (dominant species, 45–50 % of total phytoplankton abun-
solution was filtered through a cellulose paper filter (basis dance), Scendesmus acuminatus (Lagerheim) Chodat,
weight of 84 g m−2) followed by a single boiling. This Scenedesmus quadricauda (Turpin) Brébisson and
process yielded three types of solutions after extraction Scendesmus ecornis (Ehrenberg) Chodat. These four species
times of 1, 2 and 3 months, marked as 1m, 2m and 3m, were included in the subsequent analyses and the others
each with three replicates. were omitted because their abundances were too low
For the experiment, unfiltered water from a small eutro- (<1,000 cells or colonies mL−1).
phic pond was collected in late summer. The phytoplankton The initial number of S. subspicatus was 52.5±2.9×103
consisted mainly of green algae with various species of the coenobia mL−1. After 21 days, numbers of this species in
dominant genus Scenedesmus. Water with seston was placed control 1m and 2m tanks were on similar level: 51.8±2.7×
in an aquarium and kept at room temperature, aerated and 103 colonies mL−1 (control), 46.1±4.4×103 colonies mL−1
illuminated with a fluorescent lighting with a colour tem- (1m) and 49.4±5.5×103 colonies mL−1 (2m) (Fig. 1) and
perature of 6,500 K (daylight) and giving an irradiance of statistically did not differ one from the other (ANOVA;
25 μmol photons m−2 s−1 (measured by Li-Cor LI 250A c-1m, F07.348, p>0.01; c-2m, F00.357, p>0.05; 1m–
Light Meter) 12 h day−1 for 14 days. The culture was 2m, F 02.129, p> 0.05). In the tank receiving the 3-
supplied with an initial dose of phosphorus and nitrogen month extract (3m), abundances were lower at 24.8±
(0.68 mg P L−1 and 8.43 mg N L−1). 3.0×103 coenobia mL−1. Analysis of variance showed
J Appl Phycol (2013) 25:661–665 663

Straw extract added to the bottles induced changes in

conductivity, pH and concentration of dissolved organic
carbon (Table 1). The older the extract added to the bottles,
the higher the conductivity of water, and the difference
between the controls and 3m extract after exposure was
more than 100 μS cm−1. The pH values, which were rela-
tively high in the control bottles (pH 8.6–8.9) were reduced
after adding the extract and at the end of the experiment
were within the range of pH 8.4–8.5 (mean values).
However, the organic carbon content in the bottles with
added extract was nearly twice the control concentrations
and ranged from 21.8–22.7 mg L−1 (mean values).
Fig. 1 Abundance of S. subspicatus in experimental and control
bottles after 21-day exposure to various barley straw extracts (N06;
C control; 1m, 2m, 3m extracts after 1, 2 or 3 months of extraction;
triple asterisks p<0.001; one-way ANOVA) Discussion

The exposure of the tested species to different types of

that only the 3m tank differed significantly from the extract caused either no response or a reduction in popula-
control and other extract treatments (ANOVA, F0290.1, tion numbers. The 3-month extract caused a reduction in the
p <0.001 for control; F 095.3, p < 0.001 for 1m; and number of all four Scendesmus species, while in the case of
F0103.0, p <0.001 for 2m). 1-month and 2-month extracts, the response of algae was
The other species of Scenedesmus were less abundant also observed in S. quadricauda, S. ecornis and S.
at the beginning of the experiment: S. acuminatus, 4.2± acuminatus.
1.0×103 coenobia mL−1; S. quadricauda, 4.0±1.2×103 S. subspicatus proved to be resistant to 1m and 2m
coenobia mL−1; and S. ecornis, 1.4±0.4×103 coenobia extracts, but its reaction to the 3-month extract was most
mL−1. The exposure to the extracts caused species- evident among other species of the genus. Interestingly,
specific effects (Fig. 2). S. acuminatus abundances were other studies show that this species may be stimulated by
reduced only in the tanks with 3-month extract, as noted decomposing barley straw (Martin and Ridge 1999). The
above for S. subspicatus, and the difference between the authors applied wet barley straw (3–6 months of age) di-
numbers in the controls and 3m tanks was statistically rectly in doses several times higher than in our study
significant (F011.8, p <0.01). The numbers of S. quad- (4,000 g of straw m−3), but the exposure lasted only 4 days,
ricauda were statistically lower in 2m and 3m bottles which corresponds to the addition of small quantity of
(F010.3 and F050.8, respectively; p <0.01), although extract. Murray et al. (2010) studied the effect of phenolic
the population reduction was also observed in 1m bot- compounds, which are usually released from decomposing
tles (F08.5, p00.015). S. ecornis had reduced numbers barley straw on three species of algae, including two chlor-
after exposure to all types of extracts (F028.5 for 1m, ophytes. Although algistatic effects were observed in some
F022.6 for 2m and F070.6 for 3m; p <0.01). of these compounds, S. subspicatus appeared the most re-
sistant to any inhibition. There are also some reports on S.
quadricauda responses to barley straw addition. Ferrier et
al. (2005) have ranked this species among algae resistant to
barley straw extract; their experiment showed a slight

Table 1 Mean (±standard deviation) values of conductivity, pH and

concentration of total organic carbon in control samples and samples
with 1-, 2- and 3- months extracts

C 1m 2m 3m

EC (μS cm−1) 529.6±5.9 592.2±26.6 638.8±24.3 646.5±22.1

pH 8.8±0.1 8.4±0.3 8.5±0.2 8.5±0.2
Fig. 2 Abundance of S. acuminatus, S. quadricauda and S. ecornis in TOC (mg L−1) 13.2±0.7 21.8±1.0 22.2±3.2 22.7±3.7
experimental and control aquaria (N06; C control; 1m, 2m, 3m extracts
after 1, 2 or 3 months of extraction; single asterisk p<0.05; double TOC total organic carbon; C control; EC conductivity; 1m, 2m, 3m
asterisks p<0.01; triple asterisks p<0.001; one-way ANOVA) extracts after 1, 2 or 3 months of extraction
664 J Appl Phycol (2013) 25:661–665

increase in cell numbers under 2-week exposure to 2-month- suspected that they are different from those produced in later
old extract. These reports may not be in conflict with the phases (Ferrier et al. 2005).
results obtained in our experiment if we put forward a As both the rotting process and release of compounds is
hypothesis that S. subspicatus and S. quadricauda respond connected with microbial activity, physical factors, like tem-
only to long-term exposure of rotting straw. A similar view perature, should also be considered. Previous research in
is presented by Choe and Jung (2002), who found that which barley liquor was prepared in laboratory was very
certain species, although inhibition-responsive at high doses similar in terms of the temperature used. The temperature of
of plant extracts, may show an increase on exposure to extraction in experiments was stated to be between 18 and 25 °
lower extract concentrations. Our results also confirm the C or ‘room temperature’, but any attempts are known to
thesis that the inhibitory effect of straw on algae is not measure the impact of this factor on the extract activity
universal over all taxa but rather depends on particular (Gibson et al. 1990; Martin and Ridge 1999; Ball et al.
species (Brownlee et al. 2003; Ferrier et al. 2005). 2001; Terlizzi et al. 2002; Ferrier et al. 2005). Another ques-
We are aware that experiments performed on a poorly tion is the light availability during rotting of the straw. If the
controlled phytoplankton assemblage derived from a natural decomposition and release of the anti-algal compounds are
habitat, instead on pure algal cultures, may give results whose only driven by fungi, the light conditions during the extraction
interpretation is limited. In such a community, the growth and are not so significant, so the majority of extracts in mentioned
development of individual species is affected by many factors, research were conducted in the dark. However, some research-
including trophic interactions with other organisms. For ex- ers point that rotting straw can release precursive phenolic
ample, the growth of bacteria caused by an increase in dis- compounds which can undergo phototransformation in oxy-
solved organic matter derived from straw may result in genated conditions, which can result in production of hydro-
reduced availability of nutrients to algae as a result of compe- gen peroxide and other phytotoxic substances (Everall and
tition (Klug 2005). We did not monitor the nutrient concen- Lees 1997). If we take into consideration, that the use of
trations nor dynamics during the experiment, which could be barley straw should be a technique applicable in lakes and
important in the reduction of algae growth, nor did we exam- reservoirs, laboratory studies on straw extraction should mim-
ine the chemical composition of the extract that inhibits the ic field conditions (Ó hUallacháin and Fenton 2010). This
algae. The experimental results showed only that the extract implies that both temperatures and light conditions should be
addition changed some of the basic water chemistry, such as similar to those in freshwaters during vegetation season.
conductivity (increase), pH (decrease) and, most obviously, The results of the presented research showed that 3-
the content of total organic carbon (increase). However, the month extraction of barley straw produces the best results
aim of the research was to determine the importance of the in the inhibition of all species of Scenedesmus, but the
straw extraction duration for inhibiting algae; hence, the main inhibiting effect can be achieved in some species by extrac-
conclusions can only be applied to this goal. tion lasting 1 month. This confirms that the effect of dura-
Why is the extraction time important? It is generally tion of barley straw extraction on its inhibiting effect is an
accepted that algal growth inhibition by barley straw is important problem and that research on this topic should be
associated with the secretion of various chemical substances continued. They should include not only the effects of time
as algaecides, among them phenolic substances that play a but also the role of light, temperature and oxygen content, as
key role (Pillinger et al. 1994; Waybright et al. 2009). These all these factors can be fixed with microbial activity and
substances are derived from microbial degradation of the other mechanisms responsible for leaching anti-algal com-
lignin in the straw tissues, performed mainly by fungi pounds. Such knowledge can be transferred into practice in
(Pillinger et al. 1992). The most active substances begin to the water quality management in lakes and reservoirs, by the
be formed only after 3 months of straw incubation in water determination of not only the adequate doses but also the
(Pillinger et al. 1994; Ferrier et al. 2005), which is probably conditions in which the straw should be applicable.
due to the time needed by a fungal community to colonise
the substrate (Murray et al. 2010). Moreover, initially, lignin
Open Access This article is distributed under the terms of the
is not decomposed as this process occurs only under con-
Creative Commons Attribution License which permits any use, distribu-
ditions of nitrogen deficiency, which is rare in eutrophic tion, and reproduction in any medium, provided the original author(s) and
waters (Kirk and Farrell 1987). At first, intensive decompo- the source are credited.
sition of cellulose occurs in straw tissues; Murray et al.
(2010) found a total loss of cellulose in barley straw after
5 months of incubation in water. Unfortunately, there are no References
studies that have attempted to identify the chemical com-
pounds which are products of the early stage of barley straw Ball AS, Williams M, Vincent D, Robinson J (2001) Algal growth
rotting (including cellulose decomposition), although it is control by a barley straw extract. Bioresour Technol 77:177–181
J Appl Phycol (2013) 25:661–665 665

Barrett PRF, Littlejohn JW, Curnow J (1999) Long-term algal control McComas S (2003) Lake and pond management guidebook. CRC
in a reservoir using barley straw. Hydrobiologia 415:309–313 LLC, Lewis, London
Boylan JD, Morris JE (2003) Limited effects of barley straw on algae Molversmyr A (2002) Some effects of rotting straw on algae. Verh
and zooplankton in a midwestern pond. Lake Reserv Manage Internat Verein Limnol 27:4087–4092
19:265–271 Murray D, Jefferson B, Jarvis P, Parsons SA (2010) Inhibition of three
Brownlee EF, Sellner SG, Sellner KG (2003) Effects of barley straw algae species using chemicals released from barley straw. Environ
(Hordeum vulgare) on freshwater and brackish phytoplankton and Technol 31:455–466
cyanobacteria. J Appl Phycol 15:525–531 Newman JR, Barrett PRF (1993) Control of Microcystis aeruginosa by
Butler R, Reedyk S, Murrell S, Mah B (2005) Use of barley straw for decomposing barley straw. J Aquat Plant Manage 31:203–206
algal control in prairie dugouts: final report. Agriculture and Agri- ÓhUallacháin D, Fenton O (2010) Barley (Hordeum vulgare)-induced
Food Canada, Prairie Farm Rehabilitation Administration, Regina growth inhibition of algae: a review. J Appl Phycol 22:651–658
Saskatchewan Pillinger JM, Cooper JA, Ridge I, Barrett PRF (1992) Barley straw as
Caffrey JM, Monahan C (1999) Filamentous algal control using barley an inhibitor of algal growth III: the role of fungal decomposition.
straw. Hydrobiologia 415:315–318 J Appl Phycol 4:353–355
Choe S, Jung I (2002) Growth inhibition of freshwater algae by ester Pillinger JM, Cooper JA, Ridge I (1994) Role of phenolic compounds
compounds released from rotted plants. J Ind Eng Chem 8:297–304 in the antialgal activity of barley straw. J Chem Ecol 20:1557–
Everall NC, Lees DR (1997) The identification and significance of 1569
chemicals released from decomposing barley straw during reser- Ridge I, Pillinger JM (1996) Towards understanding the nature of algal
voir algal control. Water Res 31:614–620 inhibitors from barley straw. Hydrobiologia 340:301–305
Ferrier MD, Butler BR, Terlizzi DE, Lacouture RV (2005) The effects Terlizzi DE, Ferrier MD, Armbrester EA, Anlauf KA (2002) Inhibition
of barley straw (Hordeum vulgare) on the growth of freshwater of dinoflagellate growth by extracts of barley straw (Hordeum
algae. Bioresour Technol 96:1788–1795 vulgare). J Appl Phycol 14:275–280
Gibson MT, Welch IM, Barrett PRF, Ridge I (1990) Barley straw as an Vollenweider RA (1969) A manual on methods for measuring primary
inhibitor of algal growth. 2. Laboratory studies. J Appl Phycol production in aquatic environments. Blackwell, Oxford–
2:241–248 Edinburgh
Harriman R, Adamson EA, Shelton RGJ, Moffett G (1997) An assess- Waybright T, Terlizzi DF, Ferrier MD (2009) Chemical characteriza-
ment of the effectiveness of straw as an algal inhibitor in an tion of the aqueous algistatic fraction of barley straw (Hordeum
upland Scottish loch. Biocontrol Sci Tech 7:287–296 vulgare) inhibiting Microcystis aeruginosa. J Appl Phycol
Kirk TK, Farell RL (1987) Enzymatic “combustion”: the microbial 21:333–340
degradation of lignin. Ann Rev Microbiol 41:465–505 Welch IM, Barrett PRF, Gibson MT, Ridge I (1990) Barley straw as an
Klug JL (2005) Bacterial response to dissolved organic matter affects inhibitor of algal growth I: studies in the Chesterfield Canal. J
resource availability for algae. Can J Fish Aquat Sci 62:472–481 Appl Phycol 2:231–239
Martin D, Ridge I (1999) The relative sensitivity of algae to decom- Wiśniewski R (2002) Attempts to eliminate cyanobacterial blooms in
posing barley straw. J Appl Phycol 11:285–291 Lake Leasinskie. Environ Prot Eng 1:15–26