Aquaculture 416417 (2013) 374379

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Aquaculture
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Short communication

Culture of the cladoceran Moina macrocopa: Mortality associated with

agellate infection
Sarah L. Poynton a,, Philipp Dachsel b, Maik J. Lehmann c, Christian E.W. Steinberg b
a
Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Room 855 Edward D. Miller Research Building, 733 North Broadway, Baltimore,
MD 21218, USA
b
Freshwater and Stress Ecology, Institute of Biology, Humboldt University, Berlin 12437, Germany
c
Molecular Parasitology, Institute of Biology, Humboldt University, Berlin 10115, Germany

a r t i c l e

i n f o

Article history:
Received 9 July 2013
Received in revised form 16 September 2013
Accepted 17 September 2013
Available online 25 September 2013
Keywords:
Adhesive agellum
Bodoid agellate
Cladoceran
Kinetoplastid
Moina macrocopa

a b s t r a c t
Cladocerans are important food animals in aquaculture, key grazers in freshwater ecosystems, and model animals
for ecotoxicological investigations. Their epibiont community, extensively studied in Daphnia, includes lamentous bacteria, fungi, algae, peritrich ciliates, and rotifers; although epibionts are usually benign, heavy infections
can be detrimental. During our laboratory culture of female Moina macrocopa Straus, we observed a novel agellate infection associated with mortality. At day 10, all M. macrocopa were alive in uninfected cultures, whereas in
untreated infected cultures, the survival was signicantly lower: only 26% of cladocerans were alive. In infected
cultures treated with humic substances (as 25 mg L1 dissolved organic carbon), mortalities were comparable to
those in the untreated infected cultures; in contrast, in the infected cultures treated with 4 g L1 sea salt, mortalities were arrested, and 76% of the M. macrocopa were alive at day 10. Moribund cladocerans were transparent,
had empty digestive tracts, and greatly reduced motor activity. Free-swimming agellates moved forward with a
wobbling motion, rotating around their long axis; they also attached to cladoceran tissue, the Petri dish, and the
glass slide, by the tip of their posterior agellum. Flagellates preserved for scanning electron microscopy were
6.9 0.7 m long and 2.1 0.3 m wide, with a short anterior agellum (6.8 1.1 m) and long posterior agellum (14.1 1.5 m). Multi-functionality of a agellum, for locomotion and adhesion, is relatively rare, and
previously reported from genera within the Kinetoplastea, suggesting that the agellate on M. macrocopa may
belong to this group. To combat agellate mass occurrence in Moina cultures, we recommend a treatment with
4 g L1 sea salt.
2013 Elsevier B.V. All rights reserved.

1. Introduction
Cladocerans of the genus Moina, and Moina macrocopa Straus in particular, are progressively important in aquaculture and ecotoxicology.
Moina spp. are increasingly used as food for larval and post-larval rearing of crustaceans (Alam et al., 1993) and teleost sh in culture (He
et al., 2001; Ingram, 2009; Pea-Aguado et al., 2009). Due to a relatively
high protein and nutrient content, Moina spp. is a superior live food
compared to Artemia (Alam et al., 1993; Loh et al., 2012). Furthermore,
the use of freshwater zooplankton, such as M. macrocopa, may be more
convenient for feeding freshwater species than is use of saltwater
Artemia (Alam et al., 1993; Loh et al., 2012).
Although Moina is widely distributed, from temperate to tropical
regions, commercial scale quantities of this cladoceran are not easily
obtained from natural habitats (Loh et al., 2013). Mass cultivation for
live feed has been successful, and Moina tolerates low oxygen and
Corresponding author. Tel.: +1 410 502 5065.
E-mail addresses: spoynton@jhmi.edu (S.L. Poynton), philipp.dachsel@gmx.de
(P. Dachsel), maik.lehmann@hu-berlin.de (M.J. Lehmann),
christian_ew_steinberg@web.de (C.E.W. Steinberg).
0044-8486/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.aquaculture.2013.09.029

high ammonia, reproduces rapidly, and grows rapidly on a range of

food sources (Loh et al., 2013). There continues to be considerable
focus on investigating different foods for mass culture of M. macrocopa
(Kang et al., 2006; Loh et al., 2009, 2013).
In the laboratory, Moina spp., and Daphnia magna Straus are widely
used model animals in ecotoxicity testing of synthetic and natural
xenobiotics. Of particular note is that Moina sp. may be used as the
replacement for Daphnia in regions where the latter does not occur
naturally (Ferro-Filho et al., 2010; Mano et al., 2010; Sarma and
Nandini, 2006).
The successful and reliable culture of cladocerans as food for
aquaculture species is dependent on many factors, including maintenance
of healthy stocks, and effective diagnosis of disease-causing organisms
such as parasites. Cladocerans are hosts to a diversity of epibiont taxa, including lamentous bacteria, fungi, algae, peritrich ciliates, and rotifers
(Ebert, 2005; Green, 1974). Heavy coatings of epibionts can be a weight
burden, increase drag (Gilbert and Schrder, 2003), reduce population
growth (Green, 1974; Stirnadel and Ebert, 1997), and those on the thoracic limbs can lower the resistance of their host to oxygen deciency
(Pacuad, 1939). Among the parasitic taxa infecting cladocerans are bacteria, fungi, microsporidia, cestodes, and nematodes, which may cause

S.L. Poynton et al. / Aquaculture 416417 (2013) 374379

behavioral changes (Decaestecker et al., 2005; Makrushin, 2010) and reduced egg production (Green, 1974; Stirnadel and Ebert, 1997).
Although agellates have not been reported from cladocerans,
they do infect copepods, another group of small freshwater crustaceans (Hitchen, 1974), and thus they might be found on cladocerans.
Cephalothamnium cyclopum Stein (incertae sedis Kinetoplastea)
forms stalked colonies on the copepod Cyclops sp.; one agellum attached to a communally-secreted stalk, the other is used in food
gathering (Hitchen, 1974).
While the epibiont and parasite fauna of cladocerans is well known
for Daphnia (Ebert, 2005), the fauna of the increasingly important
genus Moina is little known. Since the classical study by Green (1974),
identifying a variety of epibionts and parasites on M. macrocopa, such
as Megachytrium sp., Chloranigiella epizooticum Korschikoff, Pansporella
perplexa Chatton, Epistylis helenae Green, and Brachionus rubens
Ehrenberg, there appears to have been only one report of a parasite
in Moina, namely the microsporidia Gurleya sp. in M. macrocopa
(Makrushin, 2010). We now extend knowledge of pathogenic infections in Moina spp. by reporting our light microscopy and scanning
electron microscopy observations on the dense infections of agellates associated with mortality of cultured M. macrocopa used in xenobiotic exposure experiments.

2. Materials and methods

2.1. Stress ecology studies and source of Moina
The background for the present investigation was our maintenance
of cultures of M. macrocopa for stress ecology studies, in which we
aimed to determine whether the heritage of cross tolerance was epigenetically controlled and based on DNA methylation in the presence of
humic substances. To pursue this, in xenobiotic experiments, we

375

pre-exposed Moina to humic substances, and then tested their

cross tolerance against sea salt (following Suhett et al. (2011)). During these xenobiotic experiments, some cultures became infected
with agellates, allowing us the opportunity to study them, and
their response to DOC and salt.
M. macrocopa (Fig. 1a) is a characteristic inhabitant of small, usually
ephemeral, water bodies from temperate to tropical regions, which are
often rich in dissolved organic carbon (Petrusek, 2002). Our clone was
originally isolated from a puddle in Rio de Janeiro, Brazil (ElmoorLoureiro et al., 2010), and has been successfully used since then in life
table and cross tolerance studies in stress ecology (Hofmann et al.,
2012; Suhett et al., 2011), and in a recent DNA methylation study
(Menzel et al., 2011).

2.2. Maintenance of cladoceran cultures and cross tolerance experiments

The stock culture was maintained in articial Daphnia medium
(Klttgen et al., 1994). Only neonates of the 3rd generation under identical laboratory conditions were used for the experiments. M. macrocopa
reproduces partheogenetically under stable laboratory conditions (contrasting with sexual reproduction in times of stress), and all offspring
from this asexual reproduction were female. M. macrocopa was fed
daily, ad libitum, with the coccal green algae Raphidocelis subcapitata
(Korshikov) Nygaard, Komrek, J. Kristiansen & O.M. Skulberg.
Each xenobiotic experiment was initiated with 10 replicates, in each
of which were 10 M. macrocopa in a 200 ml Erlenmeyer ask. During
the experiments, although some replicates were lost, there was always
a minimum of 8 replicates for each experiment; (thus the number of live
cladocerans expected at each time point per experiment was 80, 90 or
100, assuming no mortalities). The asks were kept in a temperaturecontrolled room at 20 1 C, and illuminated by cool white light in a
14:10 h light:dark rhythm. Every second day, the number of live and

Fig. 1. Light micrographs of Moina macrocopa and live bodonid agellates from the M. macrocopa cultures. (a) Healthy parthenogenetic Moina macrocopa female, (b) posterior part of the
second antenna of the cladoceran, arrows indicate the agellates, (c, d) two agellates showing the short whiplash anterior agellum and the long posterior/recurrent agellum, note also
the yellow-brown refractile inclusions. Scale bars = 100 m (a, b) and 10 m (c, d).

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S.L. Poynton et al. / Aquaculture 416417 (2013) 374379

dead cladocerans was recorded, and the exposure medium was exchanged. Dead individuals were removed immediately after counting.

3. Results and discussion

3.1. Infection and mortality

2.3. Infected cultures and treatments

During the experiments, we observed that while the uninfected
controls remained healthy (these were the cultures with no addition
of humic substances or salt), the agellate-infected controls suffered
mortalities.
To try to reduce subsequent mortalities, infected cultures were
treated with either: (i) humic substances (as 25 mg L1 DOC) because
they can reduce growth and survival of aquatic parasites and pathogens
(Meinelt et al., 2007, 2008), or (ii) sea salt (as 4 g L1 sea salt), because
salt is commonly used to treat ectoparasitic infections in sh; the
concentration was well within the previously documented tolerance
range of the M. macrocopa clone we studied (Suhett et al., 2011).
During the 10 day treatment period, live and dead cladocerans were
counted every second day, and dead individuals removed, as described
above.

2.4. Microscopy and video recordings

Light microscopy and video recordings were used to document
shape and motility of the agellates. To slow the agellates, a few
drops of a 1% methyl cellulose solution were added.
Surface ultrastructure of the agellates was observed by scanning
electron microscopy study of individuals retained on the tissues of the
cladocerans. Tissues were xed with 2.5% (v/v) glutaraldehyde and 2%
(w/v) paraformaldehyde in 100 mM cacodylate buffer (pH 7.4) for
30 min at room temperature. After xation, samples were rinsed
three times for 10 min with 100 mM cacodylate buffer, and dehydrated
through a graded ethanol series. After washing three times with
hexamethyldisilazane (Electron Microscopy Sciences), agellates were
coated with gold and analyzed on a LEO 1430 scanning electron
microscope. Morphometrics were determined from examination of
SEM specimens.

2.5. Data analysis

The entire lifespan of an exposure group was derived from mortality
data, and the data at each time point [% of day 0 individuals still alive]
were means for the replicates in each group. Differences between
groups over the entire 10 days, were tested for statistical signicance by the log-rank test, which was developed specically for
lifespan curves (Bioinformatics at the Walter and Eliza Hall Institute of
Medical Research (http://bioinf.wehi.edu.au/software/russell/logrank/)).
The log-rank test has previously been applied to lifespan data for
M. macrocopa (Bouchnak and Steinberg, 2014; Suhett et al., 2011).
Differences were considered statistically signicant when p b 0.05.

Table 1
Temporal changes in mortalities in four Moina macrocopa cultures. Data are percent of day
0 individuals that were alive at each time point (means SD). Each xenobiotic exposure
experiment was initiated with 10 replicates, each with 10 individual cladocerans.
Day

Uninfected
untreated

Infected
untreated

Infected
+25 mg L1 DOC

Infected
+4 g L1 sea salt

2
4
6
8
10

100
100
100
100
100

100
97.5
70.6
38.1
26.3

100
98.8
77.4
43.1
28.5

100
97.5
90.0
88.8
76.3

0.0
0.0
0.0
0.0
0.0

0.0
7.7
25.4
17.2
16.7

0.0
3.5
14.9
11.6
18.9

0.0
4.6
7.6
11.3
7.4

In the uninfected cultures, all of the M. macrocopa were alive at

day 10 (Table 1). However, in the infected and untreated cultures,
approximately half of the M. macrocopa had died after one week,
and only 26% remained alive at day 10. Survival in the infected,
untreated cultures was signicantly lower than in the uninfected cultures (p b 0.01).
In the infected cultures treated with 25 mg L1 DOC, mortalities
were high, and were not signicantly different from the infected
cultures that were not treated. In contrast, in the infected cultures
treated with 4 g L 1 sea salt, mortalities were arrested, with 76%
of the M. macrocopa alive at day 10 (Table 1). Although survival in
this treatment was signicantly higher than in the infected cultures
treated with humic substances [25 mg L 1 DOC] (p b 0.01), it was
signicantly lower than in the uninfected controls (p b 0.05).
Moribund cladocerans were transparent, their digestive tracts
were empty, and their motor activity was greatly reduced. In contrast, the healthy individuals were slightly opalescent, their digestive tracts were full of green algae, and they moved very actively.
The addition of humic substances was ineffective against the agellates affecting the Moina, although humic substances can reduce
growth and survival of some aquatic parasites and pathogens
(Meinelt et al., 2007, 2008), and increase lifespan of hosts. However,
the addition of 4 g L1 sea salt, a concentration well within the tolerance range of the clone (Suhett et al., 2011), resulted in signicantly reduced host mortality. We presume that the agellates were sensitive to
the change in osmolarity, which in turn reduced their viability, and thus
their damaging effects to the cladocerans.
3.2. Association with cladoceran tissue
There were numerous agellates inside the body cavity of the live
M. macrocopa, and agellates were also seen outside the body (Fig. 1a,
b). Although most of the agellates were attached to the host tissues
by their long posterior agellum, they could also swim freely and then
reattach to the Moina tissues. In the culture media, agellates were
usually attached by their posterior agellum to rigid structures,
such as glass slides or a Petri-dish; free-swimming agellates were
rarely observed.
3.3. Movement
When free-swimming, the agellates moved forward with a wobbling motion, frequently rotating around their long axis. The shorter
anterior agella had a whip-like action, and the longer posterior/
recurrent agella trailed (Fig. 1cd).
When attached by the tip of their long posterior/recurrent agellum,
to the cladoceran or the culture vessel, the agellates whirled around,
and the short anterior agellum was very active. The agellates could
quickly attach, detach, and reattach.
Video clips showing the agellates inside the cladoceran tissue (low
magnication), and in the culture medium (high magnication), can be
viewed online as supplementary data (see Appendix A).
3.4. Morphometrics
The morphometrics of the agellates, when viewed under the
scanning electron microscope were (minimum, maximum, mean,
standard deviation): 5.88.1 m long (mean 6.9 0.7, n = 15)
and 1.52.5 m wide (mean = 2.1 0.3, n = 12) (Fig. 2ad). The
anterior agellum was 5.48.5 m long (mean = 6.8 1.1, n =
13), and the posterior/recurrent agellum was 13.515.4 m long
(14.1 1.5, n = 8); the two agella emerged together, approximately

S.L. Poynton et al. / Aquaculture 416417 (2013) 374379

377

Fig. 2. Scanning electron micrographs of agellates from laboratory culture of the cladoceran Moina macrocopa. Note also the numerous rod-shaped bacteria. (a, b) Whole agellates showing their heterodynamic agella, the short whiplash anterior agellum and the long trailing posterior/recurrent agellum, (c, d) longitudinal view showing the anterior of the agellate,
and the two emergent agella, note also the two pores (visible in d), (e) agellate undergoing longitudinal binary ssion, which has begun at the anterior, note the two pores at the right,
and the attachment of the agellate by the agellar tip, (f) surface of agella, which is smooth, no hairs are visible [enlargement of part of panel a], and (g) surface of posterior/recurrent
agellum, longitudinal ridges appear present, no hairs are visible [enlargement of part of panel b]. Scale bars in the micrographs are 1 m.

1.0 m from the anterior end of the cell (Fig. 2c, d). The surface
of the body was smooth. In some individuals, there were two
pores each approximately 0.150.20 m in diameter, situated
1.5 m posterior to the emergence of the agella (Fig. 2d, e). In
live agellates, we observed multiple distinct yellowish-brown
refractile inclusions, approximately 0.250.50 m in diameter
(Fig. 1cd).
Individuals divided by longitudinal binary ssion, which commenced at the anterior end of the cell (Fig. 2e). In some cells, the agella

surface was smooth (Fig. 2f), while in others there were longitudinal
ridges (Fig. 2g).
In our descriptions of the agellate, we have chosen to continue
to use the term agellum for each of the locomotory organelles,
as it is a conventional practice. However, we are aware of the proposal, recently made by Adl et al. (2012), to refer to a eukaryotic agellum as a cilium, and thus such organisms as we now describe, would
be considered biciliated. It is not yet clear whether the new terminology proposed by Adl et al. (2012) will be widely adopted.

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S.L. Poynton et al. / Aquaculture 416417 (2013) 374379

3.5. Identity of the agellate

The agellate in the M. macrocopa cultures is tentatively assigned
to the Kinetoplastea on the basis of the behavior and morphology we
observed. Strong support for placement of the Moina agellate within the Kinetoplastea comes from the observation that it can attach by
the tip of the posterior agellum; this quality is unique to this group
of agellates (Vickerman, 1989).
Diverse groups of agellates contain elongate species with 2 unequal
(heterodynamic) agella, including the cryptomonads, euglenids,
kinetoplastids, and retortamonads. The unadorned surface of the
agella of organism we have described, distinguishes it from the
cryptomonds and euglenids which have hairs on the agella, and
from the retortamonds which have lamellae on the agella. Furthermore, the agellum of kinetoplastids has a unique paraagellar
rod, the presence of which may have been indicated in the Moina
agellate by the longitudinal ridges (as shown in Fig. 2g).
Within the Kinetoplastea, there are numerous genera with two agella. Based on our observations of live organisms, the agellate
we now report appears most akin to Bodo, Rhynchobodo, and
Rhynchomonas. We did not interpret the anterior of the Moina organism
as a snout (thus precluding Rhynchobodo), nor did we see creeping motility (precluding Rhynchomonas). Thus the agellate appears most similar to Bodo. However, to make a rm assignment to genus, additional
light microscopy and molecular characterization are needed, as described in the section on Recommendations below.

comparative studies, voucher specimens (infected Moina in ethanol,

and agellates preserved for light and electron microscopy), should
be deposited in museum collections.
To safeguard Moina cultures and clarify the role of the agellates in
mortality, we recommend that agellates be removed by ltering
water through a 2 m pore lter. If, however, heavy agellate infections
do occur, they may be combatted by increasing the salinity of the culture medium up to approximately 4.0 g L1, which is below the salinity
of 5.5 g L1 that is lethal to M. macrocopa (Suhett et al., 2011).
Acknowledgment
We are pleased to thank Dr. Christian Schuetz of Johns Hopkins
University School of Medicine for his assistance in translating text
from German to English. We thank Gabriele Drescher for technical
assistance with the preparation of samples for electron microscopy.
All experiments were carried out in compliance with the corresponding
laws in Germany, and the work was conducted ethically and conforms
to the uniform requirements for manuscripts submitted to biomedical
journals.
Appendix A. Supplementary data
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.aquaculture.2013.09.029.
References

3.6. Pathogenicity
The evidence suggests that the agellate was a cause, rather than a
consequence, of the poor condition of the cladocerans. In support of its
role as a pathogenic parasite is the following: (i) strong association between bodonid infection and mortality of M. macrocopa, (ii) strong association between sea salt treatment and reduced mortality, and (iii) no
association between otherwise stressed Moina populations (population
density, starvation, age) and agellate infection.
The present report appears to be the rst documentation of agellates being associated with morbidity and mortality in a population of
Moina spp. Although there are reports of the population dynamics of
Moina spp. in culture (as cited in the Introduction to this paper), and
the seasonal dynamics of M. macrocopa in natural ponds in Iran
(Khalaf and Shihab, 1979), agellate infection has not been described.
We consider it highly likely that a diversity of protozoan infections
play a role in morbidity and mortality in cultured and wild populations
of Moina spp., but that such infections are under reported.
We consider that other organisms present in the culture medium,
including bacteria and viruses, may have contributed to the morbidity
and mortality we now report. Numerous rod-shaped bacteria were
present in the cultures with the agellate-infected M. macrocopa, as
shown in the scanning electron micrographs (Fig. 2a, b).
3.7. Recommendations
To conrm the identity of agellates from infected Moina, light
microscopy and molecular approaches are needed. Light microscopy
should include staining of the cells by DAPI, which would then show
if there were large amounts of DNA in their mitochondrion (thus
conrming that they are kinetoplastids), and the location of the kinetoplast DNA (thereby allowing assignment to the Order Neobodonida,
Parabodonida, or Eubodonida) (Adl et al., 2012). Molecular characterization is also needed. Molecular tools for the detection and identication of kinetoplastids are rapidly advancing, particularly for
pathogenic taxa such as Ichthyobodo spp. (i.e. sequencing of the
SSU rDNA/18S rRNA) (Moreira et al., 2004), specic quantitative
real-time PCR targeting SSU rDNA, and novel primer sets for identication using PCR and sequencing (Isaksen et al., 2012). To facilitate

Adl, S.M., Simpson, A.G.B., Lane, C.E., Luke, J., Bass, D., Bowser, S.S., Brown, M.W., Burki, F.,
Dunthorn, M., Hampl, V., Heiss, A., Hoppenrath, M., Lara, E., Gall, L.L., Lynn, D.H.,
McManus, H., Mitchell, E.A.D., Mozley-Stanridge, S.E., Parfrey, L.W., Pawlowski, J.,
Rueckert, S., Shadwick, L., Schoch, C.L., Smirnov, A., Spiegel, F.W., 2012. The revised
classication of eukaryotes. J. Eukaryot. Microbiol. 59, 429493.
Alam, M.J., Ang, K.J., Cheah, S.H., 1993. Use of Moina micrura (Kurz) as an Artemia substitute in the production of Macrobrachium rosenbergii (de Man) post-larvae. Aquaculture 109, 337349.
Bouchnak, R., Steinberg, C.E.W., 2014. Algal diets and natural xenobiotics impact energy
allocation in cladocerans. II. Moina macrocopa and M. micrura. Limnologica 44, 2331.
Decaestecker, E., Declerck, S., De Meester, L., Ebert, D., 2005. Ecological implications of
parasites in natural Daphnia populations. Oecologia 144, 382390.
Ebert, D., 2005. Ecology, Epidemiology, and Evolution of Parasitism in Daphnia. National
Library of Medicine (US), National Center for Biotechnology Information, Bethesda
(MD).
Elmoor-Loureiro, L.M.A., Santangelo, J.M., Lopes, P.M., Bozelli, R.L., 2010. A new report of
Moina macrocopa (Straus, 1820) (Cladocera, Anomopoda) in South America. Braz.
J. Biol. 70, 225226.
Ferro-Filho, A.d.S., Soares, M.C.S., de Magalhes, V.F., Azevedo, S.M.F.O., 2010. A rapid
bioassay for detecting saxitoxins using a Daphnia acute toxicity test. Environ. Pollut.
158, 20842093.
Gilbert, J.J., Schrder, T., 2003. The ciliate epibiont Epistylis pygmaeum: selection for
zooplankton hosts, reproduction and effect on two rotifers. Freshw. Biol. 48,
878893.
Green, J., 1974. Parasites and epibionts of Cladocera. Trans. Zool. Soc. Lond. 32, 417515.
He, Z.H., Qin, J.G., Wang, Y., Jiang, H., Wen, Z., 2001. Biology of Moina mongolica (Moinidae,
Cladocera) and perspective as live food for marine sh larvae: review. Hydrobiologia
457, 2537.
Hitchen, E.T., 1974. The ne structure of the colonial kinetoplastid agellate
Cephalothamnium cyclopum Stein. J. Protozool. 21, 221231.
Hofmann, S., Timofeyev, M.A., Putschew, A., Menzel, R., Steinberg, C.E.W., 2012. Leaf litter
leachates have the potential to increase lifespan, body size, and offspring numbers in
a clone of Moina macrocopa Straus. Chemosphere 86, 883890.
Ingram, B.A., 2009. Culture of juvenile Murray cod, trout cod and Macquarie perch
(Percichthyidae) in fertilised earthen ponds. Aquaculture 287, 98106.
Isaksen, T.E., Karlsbakk, E., Repstad, O., Nylund, A., 2012. Molecular tools for the detection
and identication of Ichthyobodo spp. (Kinetoplastida), important sh parasites.
Parasitol. Int. 61, 675683.
Kang, C.K., Park, H.Y., Kim, M.C., Lee, W.J., 2006. Use of marine yeasts as an available diet
for mass cultures of Moina macrocopa. Aquac. Res. 37, 12271237.
Khalaf, A.N., Shihab, A.F., 1979. Seasonal variation in the populations of Moina macrocopa
Straus and Moina micrura Kurz (Crustacea: Cladocera) in Zoafaraniyah pools.
Hydrobiologia 62, 7577.
Klttgen, B., Dlmer, U., Engels, M., Ratte, H.T., 1994. ADaM, an articial freshwater for the
culture of zooplankton. Water Res. 28, 743746.
Loh, J.Y., How, C.W., Hii, Y.S., Khoo, G., Alan Ong, H.K., 2009. Fish faeces as a potential food
source for cultivating the water ea, Moina macrocopa. J. Sci. Technol. Trop. 5, 59.
Loh, J.Y., Ong, H.K.A., Hii, Y.S., Smith, T.J., Lock, M.W., Khoo, G., 2012. Highly unsaturated
fatty acid (HUFA) retention in the freshwater cladoceran, Moina macrocopa, enriched
with lipid emulsions. Isr. J. Aquacult. - Bamidgeh 64, 19.

S.L. Poynton et al. / Aquaculture 416417 (2013) 374379

Loh, J.Y., Ong, H.K.A., Hii, Y.S., Smith, T.J., Lock, M.M., Khoo, G., 2013. Impact of potential food sources on the life table of the cladoceran, Moina macrocopa. Isr.
J. Aquacult. - Bamidgeh 65.
Makrushin, A.V., 2010. Changes in the behaviour of Moina macrocopa (Crustacea:
Cladocera) under the inuence of Gurleya sp. (Microsporidia: Gurleyidae).
Parazitologiya 44, 475477.
Mano, H., Sakamoto, M., Tanaka, Y., 2010. A comparative study of insecticide toxicity
among seven cladoceran species. Ecotoxicology 19, 16201625.
Meinelt, T., Paul, A., Phan, T.M., Zwirnmann, E., Krger, A., Wienke, A., Steinberg, C.E.W.,
2007. Reduction in vegetative growth of the water mold Saprolegnia parasitica
(Coker) by humic substance of different qualities. Aquat. Toxicol. 83, 93103.
Meinelt, T., Schreckenbach, K., Pietrock, M., Heidrich, S., Steinberg, C.E.W., 2008. Humic
substances (review series). Part 1: dissolved humic substances (HS) in aquaculture
and ornamental sh breeding. Environ. Sci. Pollut. Res. 15, 1722.
Menzel, S., Bouchnak, R., Menzel, R., Steinberg, C.E.W., 2011. Dissolved humic substances
initiate DNA-methylation in cladocerans. Aquat. Toxicol. 105, 640642.
Moreira, D., Lpez-Garca, P., Vickerman, K., 2004. An updated view of kinetoplastid phylogeny using environmental sequences and a closer outgroup: proposal for a new
classication of the class Kinetoplastea. Int. J. Syst. Evol. Microbiol. 54, 18611875.

379

Pacuad, A., 1939. Contribution l'cologie des Cladocres. Bull. Biol. Fr. Belg. 25, 1260
(Suppl.).
Pea-Aguado, F., Nandini, S., Sarma, S.S.S., 2009. Functional response of Ameca splendens
(Family Goodeidae) fed cladocerans during the early larval stage. Aquac. Res. 40,
15941604.
Petrusek, A., 2002. Moina (Crustacea: Anomopoda, Moinidae) in the Czech Republic (a review). Acta Soc. Zool. Bohem. 66, 213220.
Sarma, S.S.S., Nandini, S., 2006. Review of recent ecotoxicological studies on cladocerans.
J. Environ. Sci. Health B 41, 14171430.
Stirnadel, H.A., Ebert, D., 1997. Prevalence, host specicity and impact on host fecundity of
microparasites and epibionts in three sympatric Daphnia species. J. Anim. Ecol. 66,
212222.
Suhett, A.L., Steinberg, C.E.W., Santangelo, J.M., Bozelli, R.L., Farjalla, V.F., 2011. Natural
dissolved humic substances increase the lifespan and promote transgenerational resistance to salt stress in the cladoceran Moina macrocopa. Environ. Sci. Pollut. Res. 18,
10041014.
Vickerman, K., 1989. Phylum Zoomastigophora, Class Kinetoplastida. In: Margulis, L.,
Corliss, J.O., Melkonian, M., Chapman, D.J. (Eds.), Handbook of Protoctista. Jones &
Bartlett, Sunbury, pp. 215238.