Available online at www.sciencedirect.

com

ScienceDirect
European Journal of Protistology 55 (2016) 75–94

Protists as bioindicators in activated sludge: Identification, ecology and

future needs夽
Wilhelm Foissner∗
University of Salzburg, Department of Ecology and Evolution, Hellbrunnerstrasse 34, A-5020 Salzburg, Austria

Available online 22 February 2016

Abstract
When the activated sludge process was developed, operators and scientists soon recognized protists as valuable indicators.
However, only when Curds et al. (1968) showed with a few photographs the need of ciliates for a clear plant effluent, sewage
protistology began to bloom but was limited by the need of species identification. Still, this is a major problem although several
good guides are available. Thus, molecular kits should be developed for identification. Protists are indicators in two stages of
wastewater treatment, viz., in the activated sludge and in the environmental water receiving the plant effluent. Continuous control
of the protist and bacterial communities can prevent biological sludge foaming and bulking and may greatly save money for
sludge oxygenation because several protist species are excellent indicators for the amount of oxygen present. The investigation
of the effluent-receiving rivers gives a solid indication about the long term function of sewage works.
The literature on protist bioindication in activated sludge is widely distributed. Thus, I compiled the data in a simple Table,
showing which communities and species indicate good, mediocre, or poor plant performance. Further, many details on indication
are provided, such as sludge loading and nitrifying conditions. Such specific features should be improved by appropriate statistics
and more reliable identification of species. Then, protistologists have a fair chance to become important in wastewater works.
Activated sludge is a unique habitat for particular species, often poorly or even undescribed. As an example, I present two
new species. The first is a minute (∼30 ␮m) Metacystis that makes an up to 300 ␮m-sized mucous envelope mimicking a sludge
floc. The second is a Phialina that is unique in having the contractile vacuole slightly posterior to mid-body. Finally, I provide
a list of species which have the type locality in sewage plants.
© 2016 Elsevier GmbH. All rights reserved.

Keywords: Activated sludge; Ciliates; Metacystis mucosa n. sp.; Phialina serranoi n. sp.

Introduction 1 kg beef. An US-American consumes about 250 l/day, a

European about 160 l, and an African only 20 l (Helmholtz-
Farming needs about two thirds of the earth’s freshwater Zentrum 2011).
and the industry takes further 23%. To produce 1 kg corn Most of the used water comes back as wastewater contam-
requires 1300 l water and 15,000 l are needed to produce inated with organic and inorganic materials. Several methods
have been developed to clean wastewater (Fig. 1) but all imi-
tate the self-purification in running waters: the wastes are
夽 Plenary lecture on occasion of the VII European Congress of Protis-
degraded by organisms, mainly bacteria, to minerals, water,
tology. This is an extended version of the conference paper from Foissner
(2014b).
CO2 , heat and new biomass, e.g. bacteria. This needs a lot
∗ Tel.: +43 66280445615. of oxygen which must be added to the wastewater treatment
E-mail address: [email protected] plants to obtain “activated = oxygenated sludge” composed
http://dx.doi.org/10.1016/j.ejop.2016.02.004
0932-4739/© 2016 Elsevier GmbH. All rights reserved.
76 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Fig. 1. Flow diagram and layout of a typical sewage-treatment work (from Curds 1992).

of about 0.5 mm-sized flocs made of inorganic and organic for the presentation of microscopic sludge analyses; and a
materials covered by a thin layer of active organisms, such as detailed table on the species and communities indicating
bacteria, fungi, and protists (Arregui et al. 2013; Curds 1992; specific parameters of plant performance.
Liebmann 1936, 1951, 1958; Mudrak and Kunst 1994; Pauli
et al. 2001; Uhlmann 1982).
The most expensive process is the oxygenation of the acti- A brief history
vated sludge. Here, protists can help to minimize oxygenation
because many of them are excellent oxygen indicators Historically, one might recognize three principal periods
(Berger et al. 1997). The activated sludge process is fre- in using protists as indicators in wastewater purification. The
quently disturbed by “sludge foaming” and “sludge bulking”, periods are connected with technical innovations in the water
that is, part of the sludge flocs does not settle and contam- works and the increasing concern of the society about the
inate the plant effluent, respectively, the river in which the heavy pollution of many rivers and lakes by organic and
effluent is discharged. Here, protists can serve as an early inorganic wastes in the industrialized countries.
and late warning tool when the river shows signs of over- The Age of Discovery and Exploitation may be set between
loading (Berger et al. 1997; Curds 1992; Ganner et al. 2002; 1914 and 1950 when Ardern and Lockett (1914) created
Liebmann 1936, 1951; Uhlmann 1982). the term “activated sludge” and researchers recognized the
This review should stimulate students and group lead- importance of protists in cleaning the wastewater during
ers to join an interesting, applied branch of protistology. It the activated sludge process (Barker 1942, 1943; Liebmann
does neither review the technical basics of wastewater treat- 1936).
ment works (for reviews see, e.g., Bayerisches Landesamt für The Age of Bloom may be set between 1950 and 2000. It
Wasserwirtschaft 1999; Uhlmann 1982; Imhoff and Imhoff started with the revision of the saprobic system by Liebmann
1993; Fig. 1) nor the important role bacteria and fungi play (1951, 1958), who recognized the usefulness of protists as
in the activated sludge process (for reviews see, e.g., Arregui indicators of water pollution when combined with metazoans
et al. 2013; Eikelboom and van Buijsen 1992; Kunst et al. and some physico-chemical parameters. These and other data
2000; Lemmer and Lind 2000). I shall concentrate on protists, were used by Curds (1966) and Curds and Cockburn (1970a,
i.e., the release from the taxonomic impediment, including b) to update bioindication in wastewater treatments plants.
the description of two new ciliates from an Austrian sewage Their faunistic and experimental studies lay the ground for
plant; brief chapters on industrial wastewaters and on indices a scientific treatment of the field. They showed which ciliate
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 77

who became employed in river pollution control soon stopped

identification work and allocated it to technical assistants!

Release from the taxonomic impediment

Whatever you want identify, plants, microscopic orga-
nisms, or beer mats, it needs good basic knowledge and some
Fig. 2. Photograph of effluents issuing from laboratory-scale pilot enthusiasm, as any work. The protists are a bit uncomfort-
plants operating in the presence (+) and absence (−) of ciliated able because they are so small that a microscope is needed.
protozoa (from Curds 1992). However, when you agree that a sewage biologist should be
an academic then the person should be able to overcome this
peculiarity, even in the time of molecular biology where clas-
species dominate in sewage plants and demonstrated their sic morphology is usually strongly neglected in the university
significance for a clear effluent (Fig. 2). education.
The increasing concern of the society on river and lake Several books are available for the identification of
pollution caused stronger government regulations and thus a microscopic wastewater organisms. For ciliates, the most
massive increase in the number of sewage-treatment works, comprehensive guide is that of Berger and Foissner (2003).
most now with nitrogen elimination (Bayerisches Landesamt It shows 357 common freshwater ciliates on flow charts,
für Wasserwirtschaft 1999; Ganner et al. 2002). Concomi- of course including those frequently occurring in activated
tantly, problems with the performance of sewage plants sludge (Fig. 4A). This guide, which is written in English
increased, such as sludge bulking and poor effluent con- language, has 160 pages and solves not only the taxonomic
taminating the rivers. This stimulated much protistological impediment but informs on several other features, such as
research on activated sludge, viz., by Curds in England, biomass, indicator value, and preferred habitat. More detailed
Fernandez-Galiano in Spain, Bick in Germany, Madoni in descriptions of the morphology and ecology of the individual
Italy, Jenkins in the U.S.A., and my group in Austria. Soon, species and many micrographs of live and silver-impregnated
the number of students and postdocs increased and hundreds specimens are contained in the monographs of Foissner et al.
of faunistic, ecological, and taxonomical papers were pub- (1991, 1992, 1994, 1995).
lished and finally collected in excellent reviews: Bayerisches The ciliate guide by Serrano et al. (2008) has a conven-
Landesamt für Wasserwirtschaft (1992, 1999), Berger and tional key to common genera in activated sludge and shows
Foissner (2003), Buck and Buck (1980), Curds (1982, 1992), 61 species each by a rather crude line drawing and a single
Eikelboom and van Buijsen (1992), Foissner and Berger micrograph. Unfortunately, the figures lack labels and any
(1996), Foissner et al. (1991, 1992, 1994, 1995, 1999), information on similar species which might cause misiden-
Ganner et al. (2002), Jenkins et al. (2004), Kinner (1984), tifications, is lacking. The real values of this booklet are the
Madoni (1991, 1994), Mudrak and Kunst (1994), Pauli et al. ecological data, i.e., under which plant conditions the species
(2001), Scherb (1968), and Sládeček (1973). have been found. It is written in English language.
During this period practical tools were developed, “Das mikroskopische Bild bei der biologischen Abwasser-
i.e., methods and indices classifying the performance of reinigung” (The microscopic picture of biological wastewater
sewage plants (Al-Shahwani and Horan 1991; Curds and purification) has been published by the Bayerische
Cockburn 1970b; Madoni 1994; Bayerisches Landesamt für Landesamt für Wasserwirtschaft (1999). Very likely, this is
Wasserwirtschaft 1992) and specific identification keys were the most comprehensive guide to the investigation of acti-
published (Berger and Foissner 2003; Berger et al. 1997; vated sludge and plant performance. It presents many good
Fernández-Galiano et al. 1996; Foissner and Berger 1996; figures of about 100 species ranging from bacteria to small
Foissner et al. 1991, 1992, 1994, 1995, 1999, Fig. 4). Thus, metazoans and other microscopic structures, such as starch
all was prepared for a successful development of the field. grains and resting cysts of protists. Further, it is the sole book
But things turned out differently. that provides help if, e.g., the sludge is bulking or foaming,
The Age of Decline commenced around the turn of the the substrate concentration is not appropriate, and indicators
century and continues. It is characterized by a decrease in of anaerobity are present.
the number of scientific papers and loss of taxonomic knowl- The colour guide to wastewater organisms by Berk and
edge, partially due to a massive change in the society but also Gunderson (1993) is a nice booklet showing a variety of orga-
due to a saturation of the field. Presently, most students and nisms ranging from bacteria to eggs of parasitic worms. All
young scientists are unwilling to learn identification of micro- are not identified to species level and thus this guide is hardly
scopic organisms and to do taxonomic work – in spite of the useable for an estimation of sewage plant performance.
good identification literature and unemployment – because An electronic guide is in preparation by Curds, Roberts,
it has low social reputation and of only vague interest in Salvado and Warren. It appears promising, showing 175 cili-
the matter. Indeed, I observed several times that colleagues ate species and their ecology, similar as in Berger and Foissner
78 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Table 1. Comparison of a high-rate wastewater treatment plant (ARA1) with conventional activated sludge plants (from Aescht and Foissner
1992).

Parameter ARA1 Conventional plants

Min Max Average Min Max Average

Temperature (◦ C) – – 30 5 25 –
pH 6.5 7.7 7 4 9 7
Oxygen content (mg/l) 0.4 4.0 2.2 1 2 –
Oxygen saturation (%) 5 50 26 – – –
Space loading (kg biochemical oxygen demand5 /m3 d) – – 21 0.25 1.5 1
Sludge age (d) – – 1 13 35 –
BOD influent (mg/l, biochemical oxygen demand) – – 21,000 – – 300
BOD effluent (mg/l) – – 3000 – – 20
COD influent (mg/l, chemical oxygen demand) – – 38,000 500 600 550
COD effluent (mg/l) – – 3400 – – 100
COD: BOD – – 1.9 – – <1.7
COD digestion (%) – – 90 – – 90
Sludge loading (kg BOD5 /kg suspended solids d) 0.7 1.0 0.8 0.05 0.3 –
Sludge volume (ml/l) – – >600 300 600 –
Sludge index (ml/g suspended solids) – – >200 80 120 –
Floc size (␮m) – – <30 150 500 –
Suspended solids (%) – – 3 – – <1
Anorganic substance (%) 45 56 50 15 65 40
Organic substance (%) 44 55 50 35 85 60
Prokaryotic biomass (mg/dry mass/l) 11,000 18,800 15,000 – – –
Eukaryotic biomass (mg/dry mass/l) 140 1470 590 – – –
Dominant bacteria Filamentous Flocculating
Dominant protozoa Flagellates Ciliates
Amoebae (number × 1000/ml) 0 770 340 0 19 –
Flagellates (number × 1000/ml) 0 5400 2000 0 17,000 2000
Globules (number × 1000/ml) 385 6000 1000 –
Ciliates (number × 1000/ml) 10 157 54 1 50 –
Rotifers (number/ml) 0 200 29 0 2300 –
Nematodes(number/ml) 0 0 0 – – 1

(2003). Further information see: http://www.nhm.ac.uk/ species level or some misidentified at all, e.g. in the study of
research-curation/projects/wastewater-ciliate/ Santos dos Araújo et al. (2014).
Aescht and Foissner (1992) investigated the organism
community of a heavily loaded (21 kg BOD5 /m3 d) plant
with pharmaceutical wastes. The organic substance on aver-
Molecular identification
age amounted to 16 g dry mass/l. The organism community
consisted of 96% bacterial and 4% protozoan biomass
A molecular identification kit for the common sludge cili-
(Table 1). Related to suspended solids the protozoan biomass
ates would minimize the problems discussed in the forgoing
constituted 2%; four tonnes protozoa (wet mass) are produced
chapter. Unfortunately, such a kit is not available although
in 1400 m3 wastewater daily. Very likely, this is the highest
the 18S rRNA sequence is available for rather many sludge
production value reported for heterotrophic protists. During
ciliates while community analyses are still in their infancy
the 1-year-study, about two flagellate species, two amoeba,
(Eisenmann et al. 2001; Guggiari and Peck 2008; Marsh et al.
eight ciliate, one nematod, and one rotifer species were rec-
1998; Matsunaga et al. 2014).
ognized. Among the ciliates, one undescribed species was
discovered, Parastrongylidium oswaldi.

Four tonnes of protozoa/day in a highly

loaded sewage plant Sludge indices
The sewage of industrial wastewaters has been poorly stud- To have a single, simple numerical value for sewage plant
ied. Often most protist species have not been identified to performance is the dream of government administrators. A
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 79

valuable, integrative method that gives information on plant

performance some days before and after the investigation.
I think progress is possible, when the identification of the
organisms is improved and a combined view is applied,
i.e., when the plant type, the structure of the flocs, the
bacterial community, and the usual physico-chemical param-
eters are taken into account (Jenkins et al. 2004; Kunst
et al. 2000; Lemmer and Lind 2000). In contrast to conven-
tional wastewater systems, Pérez-Uz et al. (2010) observed
a high number of small heterotrophic flagellates (mainly
bodonids) and small amoebae in enhanced-nitrogen removal
wastewater treatment systems. Likewise, Canals et al. (2013)
observed that ciliate diversity was much lower in wastewa-
ter with high ammonium concentration than in conventional
systems. Further, systems which include anoxic or anaero-
bic periods must be evaluated with care (Dubber and Gray
2011b).
Protists are also important indicators in the environmen-
tal water bodies that receive the plant effluent. If the plant
performance was good the past 2–3 weeks, then the water
quality should be not or only slightly changed (Liebmann
1951, 1958).

Activated sludge, an almost untouched

habitat for protist taxonomists

Fig. 3. Madoni-indices of 85 sewage plants from 38 wastewater There is a wide belief that mostly common, usually
works in Salzburg. I–IV: quality classes. From Ganner et al. (2002). polysaprobic protists occur in activated sludge. Indeed, sev-
eral of the most common sludge ciliates belong to this
domain. But there are also many less common species which
well known example is the saprobic index for evaluating the are often poorly known because they are infrequent and of
quality of running waters (Sládeček 1973). low abundance in their natural habitats. Some may get high
Five indices/methods have been developed for the per- abundances in activated sludge and thus can then be studied
formance of activated sludge plants. Most widely applied in detail, for instance, Chilodonatella minuta, Prodiscophrya
is the sludge biotic index (SBI, Fig. 3) by Madoni (1994), collini, and Enchelyomorpha vermicularis redescribed by
while the indices of Curds and Cockburn (1970b), Morishita Becares and Foissner (1994), Aescht and Foissner (1992),
(1970), Al-Shahwani and Horan (1991), and the Bayerische and Foissner and Foissner (1995).
Landesamt (1992) have been rarely and mainly locally used. Very likely, there are many undescribed protists in acti-
Both, the saprobic index and the SBI got heavy critic vated sludge plants. Some may belong to the “rare biosphere”
because it seems impossible to “reduce” the organism (Weisse 2014) and may develop to recognizable numbers
communities to a single, numerical value. However, most in activated sludge. The following ciliates have their type
users recommend such indices when they are interpreted locality in wastewater plants: Colpoda ecaudata (Liebmann,
cautiously (Arévalo et al. 2009; Arregui et al. 2013; 1936; Foissner, 1993) (a new colpodid from the sewer sys-
Drzewicki and Kulikowska 2011; Ganner et al. 2002 (Fig. 3); tem in the town of Leipzig, Germany); Epistylis epibioticum
Liebmann 1951; Pedrazzani 2014; Sládeček 1973; Toman Banina, 1983; E. longicaudatum Banina, 1983; E. poleneci
2002). saprobicum Banina, 1983 (three new peritrichs from sewage
plants in Russia); Gastronauta aloisi Oberschmidleitner
and Aescht, 1996 (a new cyrtophorid from a plant with
Sewage plant performance by bioindication mixed domestic and industrial wastewater near to the town
of Asten, Upper Austria. Also found by Silva and Silva-
Table 2 is a more complete version of a Table first put Neto (2001) in a sludge plant in Rio de Janeiro, Brazil
together by Arregui et al. (2013). It shows, inter alia, that but misidentified as G. membranaceus; see their Figs. 46,
sewage plant performance by bioindication is possible to a 47); Deviata brasiliensis Siqueira-Castro, Paiva and Silva-
certain extent but it is not the philosopher’s stone. It is a Neto, 2009 (a new hypotrich from a sewage plant near
80 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Fig. 4. (A) shows a flow chart from the ciliate guide by Berger and Foissner (2003). (B–D) show characteristic ciliate communities (from
Berger and Foissner 2003).
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 81

Table 2. Microfaunal species and communities as indicators of sludge plant performance in conventional systems.

Organisms Performance Remarks Literaturea

Species (usually when subdominant at least)

Acineria uncinata (Fig. 4B) Mediocre Instability of sludge (due to high abundance of 19
free bacteria and insufficient cleaning; tolerates
low oxygen concentration). Present with high
densities and occurrence in nutrient removal
systems
Aspidisca cicada (Fig. 4B) Good Stable plant conditions. Good removal of BOD5 1, 19
and COD, low oxygen and nitrate 20, 25
Chilodonella uncinata Good Good effluent quality, low oxygen concent 19, 25
(Fig. 4B)
Coleps hirtus Good Effective nitrification with good removal of 1
ammoniacal-N in effluent
Enchelyomorpha Poor Microaerobic; overloading; hydraulic problems 2, 3
vermicularis (Fig. 4C)
Euplotes aediculatus Mediocre This species can be used for evaluating the 19
(Fig. 4B) toxicity of waters polluted by nickel
Euplotes mutabilis – Highly tolerant for heavy metals 22
Euplotes patella (Fig. 4B) Mediocre When abundant and in connection with many 2, 6
rotifers indicative for an increasing sludge volume
index; otherwise underload
Holophrya discolor Mediocre Microaerobic; intermitent and very low 4, 20
oxygenation; high N-reduction
Litonotus lamella Poor Deficient sludge setting 19
Litonotus obtusus Poor Poor sludge setting 16
Metopus spp. (Fig. 4C, D) Poor Anaerobic conditions; overloading; hydraulic 1, 2, 5, 9
problems
Opercularia articulata – Highly tolerant for NaCl 21
Opercularia coarctata Poor Low effluent quality and toxicity (associated with 19
(Fig. 4B) high BOD5 in the effluent)
Plagiocampa rouxi Mediocre Microaerobic; intermitent and very low 4, 20
oxygenation; high N-reduction
Spirostomum teres (Fig. 4D) Mediocre Microaerobic; intermitent and very low 4, 19
oxygenation; high N-reduction. Toxicity by heavy
metals, pesticides, or phenols
Stylonychia mytilus – Removes Pb 2+ from sludge 22
Tetrahymena pyriformis – Poor Related to short sludge retention time and 19
complex colonization processes. Good indicator of
polysaprobic to isosaprobic conditions
Trimyema compressum Poor Microaerobic; overloading; hydraulic problems 2, 3
(Fig. 4C)
Trithigmostoma cucullulus Good Sensitive to ammonia and phosphate 20
Trochilia minuta Good Indicative of good nitrifying conditions; underload 19
Uronema nigricans Poor Low effluent quality 19
Vorticella campanula Good High effluent quality; underload 2, 10, 25
(Fig. 4A)
Vorticella convallaria Mediocre Lack of nitrification 1
(Fig. 4B)
Vorticella convallaria and Good High sludge retention time; underload 2, 16, 17
Arcella hemisphaerica
Vorticella Poor Low clearing efficiency, especially when 7, 8, 9, 17,
microstoma/infusionum connected with high flagellate abundance; 25
(Fig. 4D) and Opercularia anaerobic; high sludge load and sludge volume
sp. (Fig. 4B) index; low oxygen concentration
Vorticella striata – complex Poor Poor effluent quality and insufficient cleaning 1, 19
(positive relation with high volumetric load, mass
load and high effluent BOD5 ).
Zoothamnium procerius Good Good depuration efficiency 19
82 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Table 2. (Continued)

Organisms Performance Remarks Literaturea

Communities (when dominant or subdominant)

Small flagellates Poor Low effluent quality; oxygen depletion; overloading; 8, 18, 19
sludge maturation period; onset of nitrification
Small naked amoebae Poor Very high load; not easily degradable material; 8, 19
sludge maturation. Associated to anaerobic
conditions and processes of high load poor yields in
CAS. Small and large naked amoebae are compatible
with good nitrification when WWTP are designed for
biological nutrient removal. >50 ␮m are present in
low organic loading rates and moderate and higher
SRT, common in nutrients removal WWTP
Small flagellates, naked Poor Unstable sludge; sludge maturation; toxic influences 2, 9
amoebae, swarmers of
peritrich ciliates; many
dispersed bacteria
Testate amoeba, e.g., Arcella, Good Underloading, high sludge retention time; usually 8, 19, 23
Euglypha found in N-removal plants. Good performance
related to low mass load, sufficient oxygenation and
good nitrification
Testate amoebae; crawling Good Healthy, low-loaded, sufficiently aerated and 12
ciliates; attached peritrich well-flocculated sludge with high effluent quality
ciliates with width
peristome; nematods;
rotifers
Glaucoma, Dexiostoma Poor Insufficient oxygenation; many dispersed bacteria; 2, 9
campylum (Fig. 4D), poor effluent quality
Vorticella microstoma and
peritrich swarmers,
flagellates and naked
amoebae
Vorticella infusionum Poor High-loaded with insufficient oxygen; shock-load; 12
(Fig. 4D); Opercularia high ammonia; many dispersed bacteria
coarctata (Fig. 4B);
Acineria uncinata
(Fig. 4B); Small flagellates
Spirostomum minus, Euplotes – Indicative of 80–85% nitrogen removal and of 24
affinis, Opercularia 90–95% of carbon compounds; do not tolerate NH4 +
coarctata concentrations above 14 mg/l
Heterotrich ciliates (Fig. 4D) Poor Poor operation of RBC system 5
and many flagellates
Epistylis (Fig. 4B), large Good When in last stage of RBC system 13
naked amoebae, rotifers
Green algae on plant wall Good Underload since a long time 9
Small swimming ciliates Mediocre Too short sewage retention time; insufficient 8, 19
oxygenation. Positive relationship with effluent
BOD5 and negative correlation with sludge age
Large swimming ciliates Mediocre Overloading; insufficient oxygenation 8
(Fig. 4B, D)
Crawling ciliates (abundance Good Sludge volume index <200 8
>2000/ml)
Sessile and crawling ciliates Good 8
Crawling and attached ciliates Good High ratio indicates good effluent. 15
Sessile ciliates Good Transient phenomena, such as recent sludge 1, 8, 19
extraction, discontinuous load.
Good management and performance of plant.
Positively related to volumetric load and negatively
to effluent BOD5
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 83

Table 2. (Continued)

Organisms Performance Remarks Literaturea

Ciliates Good When abundance is 106 /l or more 8

Ciliates – Abundance <104 /l (poor), 104 –106 /l (mediocre), 11, 17
>106 (good)
Metopetum (Fig. 4C) Poor Anaerobic conditions; overload; hydraulic 2, 9
problems; putrefaction
Swimming and attached Mediocre When highly diverse indicative for stable sludge 2, 9
ciliates but insufficient effluent quality
Swimming ciliates Mediocre Often dominate in plants with short retention 1, 18
time; effluent mediocre; disappear after pH-shock
Vorticella Good Well-setting sludge 10
microstoma/infusionum
(Fig. 4D) and V. campanula
Cyrtophorids, hypotrichs, Good Good operation of RBC system (Rotation 5
scuticociliates, Biological Contactor)
pleurostomatids (Fig. 4B)
Opercularia, Uronema, Poor Indicate overloading when in last stage of RBC 13
nematods system
Carnivorous ciliates, e.g., Poor Poor-setting sludge 10
Litonotus lamella,
Amphileptus
Aspidisca cicada (Fig. 4B), – High sludge retention time 10
Chilodonella spp.
(Fig. 4B), Vorticella striata
Epistylis plicatilis and Decreasing Indicate beginning sludge bulking when their 14
Vorticella striata abundances distinctly increase; high sludge
volume index (SVI)
a 1 = Martin-Cereceda et al. (1996), 2 = Foissner et al. (1995), 3 = Pérez-Uz et al. (1998), 4 = Ganner et al. (2002), 5 = Martín-Cereceda et al. (2001),

6 = Cingolani et al. (1991), 7 = Gori et al. (1991), 8 = Madoni (1994), 9 = Bayerisches Landesamt für Wasserwirtschaft (1992, 1999), 10 = Lee et al. (2004),
11 = De Marco et al. (1991), 12 = Drzewicki and Kulikowska (2011), 13 = Berri and Casaschi (1991), 14 = Hu et al. (2013), 15 = Bedogni et al. (1991), 16 = Zhou
et al. (2006), 17 = Toman (2002), 18 = De, Cybis and Horan (1997), 19 = Arregui, Liébana, Calvo, Pérez-Uz, Salvado and Serrano (2013), 20 = Dubber and
Gray (2011a), 21 = Salvadó et al. (2001), 22 = Rehman et al. (2006), 23 = Chen et al. (2004), 24 = Luna-Pabello et al. (1996), 25 = Čech et al. (1994).

to the town of Rio de Janeiro, Brazil); Euplotes mus- Future needs

cicola lahorensis Chaudhry and Shakoori, 2012 (a new
subspecies from industrial wastewater in Pakistan); Meta- Wastewater technology has reached a high standard. When
cystis galiani Arregui et al., 2010 (a new prostome from biologists want not disappear from the wastewater industry,
the wastewater of a health resort in Valencia, Spain); Meta- they should improve (i) the identification of the organisms and
cystis mucosa (see description below); Parastrongylidium redescribe or describe insufficiently known or new species,
estevesi Paiva and Silva-Neto, 2005 (a new hypotrich from (ii) the knowledge on the ecology of the individual indi-
a sewage plant in Brazil); Parastrongylidium oswaldi Aescht cator species, (iii) the knowledge on plant technology and
and Foissner, 1992 (a hypotrich discovered in a heav- statistics, (iv) data interpretation by incorporating plant type,
ily loaded pharmaceutical sewage plant in Tyrol, Austria); physico-chemical parameters and the bacterial community,
Parentocirrus brasiliensis Paiva and Silva-Neto, 2004 (a (v) the molecular identification of sludge protists by inventing
new hypotrich from a sewage plant in Rio de Janeiro, methods that can be handled by technicians.
Brazil); Phialina serranoi (see description below); Propy-
xidium elongatum Shizheng and Zhengxue, 1995 (a new
peritrich from paper mill sludge in China); Telotrochidium Description of new species
matiense (Martín-Cereceda et al., 1996) Martin-Cereceda
et al., 2007 (a new, stalkless peritrich from a sewage plant Both species are from an oxygenated, well working
in Madrid, Spain); Vorticella aerotenci Banina, 1983; V. wastewater plant in Austria (see type locality and Ganner et al.
geispicae Banina, 1983; V. hyalina Banina, 1983; V. peter- 2002). They were discovered during wastewater courses in
hoffi (four new Vorticella species from sludge plants in 1993 and 1999. One species is outstanding because it makes
Russia); and Trochiliopsis australis Foissner et al., 1988 a voluminous mucous coat thus appearing like a sludge floc.
(a new microthoracid from a sludge plant in southern For ciliate preparation methods and terminology, see Foissner
Australia). (2014a) and Lynn (2008).
84 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Metacystis mucosa nov. spec. (Figs 5A–M, 6A–L, 7A–P; (reg. no. 836/2015–838/2015) with Chatton–Lwoff sil-
Table 3) ver nitrate-impregnated cells, and four slides (reg. no.
Diagnosis: Lives in a mucous coat up to 300 ␮m across. 839/2015–842/2015) with Klein–Foissner silver nitrate-
Size in vivo about 40 × 17 ␮m, contractile up to half of impregnated specimens have been deposited in the Biology
body length. Extended specimens lageniform or bursiform, Centre of the Upper Austrian Museum in Linz (LI). The holo-
contracted cells barrel-shaped or ovate. With a conspicuous type and other relevant specimens have been marked with
posterior vacuole. Macronucleus in posterior half, globular. black ink circles on the coverslip.
Contractile vacuole in or near mid-body. Lorica lageniform, Description: Metacystis mucosa has an ordinary variabil-
made of a thin, colourless membrane. Cortex and silverline ity with all important features having variation coefficients
pattern as typical for genus. On average 31 longitudinal cil- ≤ than 15%.
iary rows forming eight transverse ciliary rings; caudal cilium Body size difficult to study because contractile by up
about as long as body. On average nine circumoral dikinetids to 50%. Several measurements suggest (Table 3): size of
and five perioral ciliary rings composed of monokinetids. extended specimens in vivo 35–50 × 15–20 ␮m, usually
Type locality: Activated sludge plant for the town about 40 × 17 ␮m; slightly smaller (mean: 34 × 12.8 ␮m,
of Seekirchen, Salzburg county, Austria, 47◦ 53 8.4 N respectively, 36 × 14 ␮m) after Chatton–Lwoff silver
13◦ 7 14.26 E. nitrate impregnation when 5% preparation shrinkage are
Etymology: The Latin adjective mucosa refers to the volu- added; strongly shrunken in protargol preparations (mean
minous mucous coat. 27 × 10 ␮m). Body shape also highly variable because very
Type material: The holotype slide (reg. no. 832/2015) different in extended and contracted state. When extended
and three paratype slides (reg. no. 833/2015–835/2015) elongate bursiform or lageniform with a length: with ratio
with protargol-impregnated specimens, three paratype slides of 2.0–3:1 and a slender more or less curved neck and a

Table 3. Morphometric data on Metacystis mucosa.

Characteristicsa Method Mean M SD SE CV Min Max n

Body, length IV 39.3 37.0 5.5 2.0 14.1 35.0 50.0 8

CHLE 34.0 33.0 4.5 1.0 13.4 27.0 44.0 21
CHLC 21.2 21.0 4.3 0.9 20.2 17.0 34.0 21
PE 26.5 27.0 2.9 0.6 11.0 22.0 31.0 21
PC 14.6 14.0 1.8 0.4 12.5 12.0 18.0 21
Body, width IV 16.8 16.0 2.1 0.7 12.3 15.0 19.0 8
CHLE 12.8 13.0 1.6 0.3 12.4 10.0 16.0 21
CHLC 16.5 16.0 2.9 0.6 17.3 13.0 24.0 21
PE 10.2 10.0 0.8 0.2 8.0 9.0 12.0 21
PC 11.5 11.0 1.5 0.3 13.1 9.0 15.0 21
Body length: width, ratio IV 2.4 2.4 0.4 0.1 15.5 1.8 3.0 8
CHLE 2.7 2.6 0.3 0.1 10.0 2.3 3.3 21
CHLC 1.3 1.3 0.1 0.1 8.0 1.1 1.4 21
PE 2.6 2.6 0.2 0.1 7.6 2.3 3.0 21
PC 1.3 1.3 0.1 0.1 7.5 1.0 1.4 21
Anterior body end to macronucleus P 9.3 9.0 2.5 0.5 26.3 5.0 14.0 21
Oral bulge, height P 2.1 2.0 – – – 2.0 3.0 20
Anterior body end to last ciliary ring P 18.6 19.0 2.2 0.5 11.9 15.0 22.0 20
Macronucleus, length P 6.2 6.0 0.6 0.1 10.0 5.0 7.0 21
Macronucleus, width P 6.1 6.0 0.6 0.1 9.8 5.0 7.0 21
Micronucleus, length P 1.6 2.0 – – – 1.0 2.0 8
Micronucleus, width P 1.3 1.0 – – – 1.2 2.0 8
Circumoral dikinetids, number P 9.0 9.0 0.6 0.2 6.4 8.0 10.0 7
Perioral ciliary rings, number P 5.0 5.0 0.0 0.0 0.0 5.0 5.0 8
Somatic ciliary rings, number P 8.0 8.0 0.3 0.1 4.1 7.0 9.0 20
CHL 8.1 8.0 0.6 0.1 7.9 7.0 9.0 22
Longitudinal ciliary rows, number P 31.3 31.0 1.8 0.5 5.7 28.0 34.0 13
CHL 31.2 32.0 2.0 0.5 6.3 26.0 33.0 13
a Data based on environmental, partially selected specimens. Measurements in ␮m. CV, coefficient of variation in %; CHL, Chatton–Lwoff silver nitrate

impregnation; CHLC, more or less contracted, silver nitrate-impregnated specimens; IV, in vivo; M, median; Max, maximum; Mean, arithmetic mean; Min,
minimum; n, number of cells investigated; P, protargol impregnation; PC, more or less contracted, protargol-impregnated specimens; PE, more or less extended,
protargol-impregnated specimens; SD, standard deviation; SE, standard error of arithmetic mean.
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 85

Fig. 5. A–M. Metacystis mucosa from life (A–E, G) and after protargol impregnation (F, H–M). A: An extended, 40 ␮m long specimen within
its mucous coat the opening of which is lined by the membranous lorica. B: A freely motile, contracted specimen, length 23 ␮m, showing
the main organelles, especially the contractile vacuole and the long caudal cilium. C: Cortex. D: An extended specimen, showing the perioral
ciliary waves. E: Cortex structure; cilia in mid of figure not shown. F: An extended specimen, showing the receptaculum. G: Formation of the
posterior vacuole. H: Basal body pattern of a slightly contracted specimen. I: brush fibres? J: Cortex structure and caudal cilium. K: Excretory
pore and macronucleus. L: Slightly oblique anterior polar view, showing the oral basal body (ciliary) pattern. M: Holotype specimen, length
22 ␮m. A, alveolus; B, bacteria; BF, brush fibres? C, cytoplasmic crystal; CC, caudal cilium; CK, circumoral kinety; CV, contractile vacuole;
EP, excretory pore; L, lorica; LI, lipid droplet; MA, macronucleus; MI, micronucleus; OB, oral bulge; PO, perioral ciliature; R, receptaculum;
V, posterior vacuole. Scale bars 10 ␮m (B, F, H–M) and 30 ␮m (A).
86 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Fig. 6. A–L. Metacystis mucosa from life, bright field (A–H) and interference contrast (C, inset; I–L) micrographs. A: A slightly contracted
specimen in a large slime floc. Note the characteristic posterior vacuole. B: An extended, swirling specimen, showing the receptaculum and
the contractile vacuole slightly posterior to mid-body. C: A contracted specimen (arrow) within a homogenous mucous coat (arrowheads). The
inset shows a cell with a conspicuous posterior vacuole. D, E: A contracted and a moderately contracted specimen forming the mucous coat.
F: An extending specimen in a large slime floc. G: An extended, feeding specimen with lorica destroyed by the preparation. H: A moderately
contracted specimen. I: A slightly pressed specimen leaving the lorica through the posterior end. The lorica is very thin and colourless. J:
A specimen showing the receptaculum. K: A strongly pressed specimen, showing compact food vacuoles and the contractile vacuole near
mid-body. L: Surface view, showing the cortex pattern and the transverse basal body (ciliary) rings (arrowheads). CV, contractile vacuole;
FV, food vacuoles; L, lorica; OC, oral cilia; R, receptaculum; V, posterior vacuole. Scale bars 10 ␮m (J–L), 15 ␮m (H), 30 ␮m (D, E, G, I),
40 ␮m (B), 50 ␮m (A, F), and 100 ␮m (C).
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 87

Fig. 7. A–P. Metacystis mucosa, basal body (∼ciliary) pattern after protargol impregnation (A–K, M) and after Chatton–Lwoff (N, O) and
Klein–Foissner (L, P) silver nitrate impregnation. A, B, J: More or less extended specimens, showing the ciliary pattern on average composed
of 31 meridional ciliary rows forming 8 transverse ciliary rings. The pore of the contractile vacuole is in or slightly posterior to mid-body
while the macronucleus is in the posterior third of the cell. C–E, I: Moderately (C) and strongly (D, E, I) contracted specimens are bursiform
(C), barrel-shaped (D), ellipsoid (E), or ovate (I). Note the long caudal cilium in (I). F, H: Posterior polar views, showing the circular shape
of the ciliary rings and the basal body of the caudal cilium. G, K: Oblique anterior polar views, showing the dikinetids of the circumoral
kinety and the monokinetids of the five perioral ciliary rings. L, P: Silverline pattern with docked extrusomes marked by arrows in (P). M:
A contracted specimen with a large posterior vacuole having attached some argyrophilic material (arrow). N, O: Cilia or kinetodesmal fibres
impregnate with the Chatton–Lwoff silver nitrate method. Note the basal body of the caudal cilium and the excretory pore between the fourth
and fifth ciliary ring (O). C, cilia (basal bodies); CC, caudal cilium; CK, circumoral kinety (dikinetids); EP, excretory pore of contractile
vacuole; MA, macronucleus; PO, perioral ciliature; SK, somatic kineties; V, posterior vacuole. Scale bars 5 ␮m (G, K, P), 10 ␮m (C–F, H–J,
L, M, O), and 15 ␮m (A, B, N).
88 W. Foissner / European Journal of Protistology 55 (2016) 75–94

broad posterior quarter (Figs 5A, D, F, H, 6B, G, 7A, B, J; an average of nine circumoral dikinetids (Figs 5L, 7G, K;
Table 3); unflattened (Fig. 7F, H). More or less contracted Table 3) with about 3 ␮m long cilia forming conical “oral
specimens bursiform (Figs 5K, F, 6A, E, 7C), barrel-shaped flaps” as in prostomatid ciliates (Foissner et al. 1999). Five,
(Figs 5B, J, 6D, 7D, O), ellipsoid (Figs 5I, J, 6C, H) or ovate rarely only four very narrowly spaced, monokinetal, peri-
(Figs 5K, 7I, M, N); about 23 × 17 ␮m in size (Table 3). oral ciliary rings with 8–10 ␮m long cilia making nice
Macronucleus usually in posterior half or third of cell, metachronal waves when swirling bacteria into the oral open-
globular to broadly ellipsoid, about 7 ␮m across in vivo; ing (Figs 5A, B, D, L, M, 6A, E, 7B, J, G, K, L; Table 3).
with distinct nucleoli about 2 ␮m in size. Micronucleus Occurrence and ecology: As yet found only at type local-
near or attached to macronucleus, broadly ellipsoid, about ity. Metacystis occurs in limnetic and marine habitats, and
2.5 × 1.5 ␮m (Figs 5A, H, M, 7A, D, E, J, M; Table 3). Arregui et al. (2010) discovered a new species attached to
Contractile vacuole usually between fourth and fifth somatic activated sluge flocs in a Spanish wastewater plant. About
transverse ciliary ring, i.e., in or near mid-body, about half of the species possibly lack a lorica while the loricate
3 ␮m across in vivo, excretory pore about 1 ␮m in diameter species are attached to a firm substrate (Arregui et al. 2010;
(Figs 5A, B, K, M, 6B, D, K, 7A, C, O). Posterior vacuole not Kahl 1930). I suppose that the enormous mucous coat of M.
contractile, one third usually within cell, two thirds projecting mucosa simulates a sluge floc, i.e., making the cell indepen-
(Figs 5A, B, 6A, C inset, E, J, 7A, E, I, N). Very likely bursts dent from wastewater flocs which tend to break or become
occasionally because it can disappear and is rebuilt (Fig. 5G). coated by filamentous bacteria.
Vacuole contents hyaline, frequently some fluffy material Species comparison: According to the reviews by Kahl
attached to inner side of vacuole membrane (Figs 5B, 7M). (1930) and Arregui et al. (2010), M. mucosa is similar to
Cortex very flexible, of typical Metacystis structure, i.e., with M. recurva Penard, 1922; M. exigua Penard, 1922; and M.
series of longitudinal and transverse ridges, forming oblong, tesselata Kahl, 1926. However, none has such a voluminous
rectangular, comparatively large alveoli with cilia inserted slime coat as M. mucosa, which is thus an important
in mid (Figs 5B, C, E, J, 6L); alveolar pattern recognizable diagnostic feature.
in vivo (Fig. 6L) and, occasionally, also in protargol- Metacystis recurva differs from M. mucosa not only by the
impregnated specimens (Figs 5J, 7B). Silverline pattern absence of a slime coat but also by body length (∼50 ␮m vs.
very similar to cortex pattern, docked extrusomes appear as ∼40 ␮m), the number of transverse ciliary rings (∼15 vs. 8),
minute rings in the transverse ridges (Fig. 7L, P); extrusomes and the number of longitudinal ciliary rows (20 vs. 31). Meta-
in vivo not recognized possibly because only 1 ␮m long and cystis exigua, a species with a mucous coat, differs from M.
of very similar refractivity as cortex. Cytoplasm colourless, mucosa by body length (13–20 ␮m vs. ∼40 ␮m), the number
usually crammed with food vacuoles, crystals, and granules of transverse ciliary rings (∼4 vs. 8), the posterior vacuole
(Figs 5A–C, 6A, D, E, G, I–K). Food vacuoles 3–4 ␮m across (absent vs. present), and the mucous coat (a few micrometre
in vivo, very likely contain compacted bacteria. Crystals vs. >100 ␮m thick). Freely motile cells of M. mucosa highly
scattered throughout body, 0.2–3 ␮m in size, sometimes in resemble M. tesselata. However, M. tesselata possibly lacks a
yellowish vacuoles about 2 ␮m across. lorica and a slime coat, is only slightly (vs. distinctly) contrac-
Lives in a highly flexible lorica difficult to recognize tile, and has about 15 (vs. 8) transverse ciliary rings formed
because very thin, colourless, and very near to the cell by about 20 (vs. 31) longitudinal ciliary rows.
(Figs 5A, 6A–C, F, G, I). Lorica possibly open at both ends, Metacystis galiani, which Arregui et al. (2010) discovered
shape as described for the extended body, extends through in an aerobic wastewater treatment plant differs from M.
the mucous coat, does not stain with alcian blue. Coat up mucosa by having a conspicuous, annulated lorica and by the
to 300 ␮m in size, stains with alcian blue indicating acid absence of both, a caudal cilium and a mucous coat. Further,
mucopolysaccharides, very hyaline often becoming recog- it has two perioral ciliary girdles composed of an anterior
nizable mainly due to attached bacteria (Figs 5A, 6A–C, F). ring of dikinetids and four posterior rings of monokinetids.
Most cells leave coat when transported to the microscope
slide but soon begin to secrete a new coat (Fig. 6D, E). Phialina serranoi nov. spec. (Figs 8A–K, 9A–I; Table 4)
Somatic cilia only 5–6 ␮m long in vivo, form an average Diagnosis: Size in vivo and in protargol preparations
of 31 very narrowly and equidistantly spaced, longitudinal about 85 × 27 ␮m when extended. Elongate ellipsoid with
rows with cilia ordered so strictly that seven to nine, oral bulge and head rather distinctly set off from trunk;
usually eight transverse somatic and five perioral ciliary broadly ellipsoid when fully contracted. Macronucleus and
rings originate. Caudal cilium about as long as body, micronucleus broadly ellipsoid. Contractile vacuole far sub-
inserts between cell and posterior vacuole, turned around terminal. Two types of rod-shaped extrusomes attached to
posterior body region in loricate cells, easily overlooked oral bulge: type I about 12 ␮m long, type II 2.5–3 ␮m. On
(Figs 5A, B, D, J, 7H, L, N, O; Table 3). average 15 ciliary rows each with one to four dikinetids
Oral opening in centre of anterior end, broadly ellipsoid anteriorly.
and about half as width as body, guides into a large, con- Type locality: Activated sludge plant for the town
ical receptaculum with a membranous structure moving to of Seekirchen, Salzburg county, Austria, 47◦ 53 8.4 N
and fro (Figs 5A, B, F, 6B, J). Oral opening surrounded by 13◦ 7 14.26 E.
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 89

Fig. 8. A–K. Phialina serranoi from life (A–C, E, F) and after protargol impregnation (D, G–K). A: Overview of a representative specimen,
showing the most important feature of this new species, viz., the far subterminally located contractile vacuole; length 85 ␮m. B, C: Type I
and type II extrusomes, length 12 ␮m and 3 ␮m. D, I, J: Details of oral bulge and head. E: Cytoplasmic inclusions. The conical structures are
possibly spines for the resting cyst. F: Optical section, showing cortical granulation. G, H: Basal body (ciliary) pattern of holotype specimen,
length 80 ␮m. K: A contracted specimen with two macronuclear nodules, very likely a post-conjugate. CG, cortical granules; CK, circumoral
kinety; CV, contractile vacuole; DK, dikinetids; E, extrusomes; EB, extrusome bundle; F, fibrous cylinder; H, head; MA, macronucleus; MI,
micronucleus; OB, oral bulge; T, trunk. Scale bars 20 ␮m (K) and 30 ␮m (A, G, H).

Table 4. Morphometric data on Phialina serranoi.

Characteristicsa Mean M SD SE CV Min Max n

Body, length 72.3 70.0 8.4 1.8 11.7 58.0 87.0 21

Body, width 23.0 23.0 2.3 0.5 9.9 18.0 27.0 21
Body length: width, ratio 3.2 3.1 0.4 0.1 12.0 2.7 4.4 21
Anterior body end to macronucleus, distance 35.7 34.0 6.3 1.4 17.7 26.0 46.0 21
Macronucleus, length 13.8 14.0 2.1 0.5 15.2 9.0 17.0 21
Macronucleus, width 9.4 10.0 1.9 0.4 20.3 5.0 12.0 21
Micronucleus, length 1.7 2.0 – – – 1.0 2.0 7
Micronucleus, width 1.3 1.0 – – – 1.0 2.0 7
Somatic ciliary rows, number 15.0 15.0 0.6 0.1 4.2 14.0 16.0 21
Monokinetids in a ciliary row, number 25.0 25.0 4.4 1.0 17.4 17.0 35.0 21
Dikinetids at begin of ciliary rows, number 2.6 3.0 0.7 0.2 25.6 2.0 4.0 21
Head, hight 4.2 4.0 0.5 0.1 12.2 3.0 5.0 21
Head, width 5.1 5.0 0.7 0.2 13.3 4.0 7.0 21
a Data based on environmental, protargol-impregnated, extended specimens. Measurements in ␮m. CV, coefficient of variation in %; M, median; Max,

maximum; Mean, arithmetic mean; Min, minimum; n, number of individuals investigated; SD, standard deviation; SE, standard error of arithmetic mean.
90 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Fig. 9. A–I. Phialina serranoi from life (A–D) and after protargol impregnation (E–I). A, B: Two pressed specimens, showing the far
subterminal contractile vacuole, the most important feature of this new species. C: Oral bulge, showing type I (arrow) and type II extrusomes
(arrowhead). D: A squashed specimen studded with conical inclusions, very likely spines for the resting cyst. E, F: Basal body (ciliary) pattern
of a paratype specimen and of the holotype. All ciliary rows begin with two to four dikinetids and scattered dikinetids occur throughout the
rows, especially in posterior region. G, H: Details of anterior body region. The cilia are very narrowly spaced in the oblique head kineties
and the last basal body of each kinety is slightly separate. I: A contracted specimen with two macronuclear nodules and thus very likely
a late post-conjugate. CG, cortical granules; CI, cilia; CR, ciliary rows; CV, contractile vacuole; DK, dikinetids; E, type I (A) and type II
(G, H) extrusomes; EB, type I extrusome bundle; F; fibrous cylinder; FV, food vacuoles; H, head; LI, lipid droplets; M, mitochondria; MA,
macronucleus; OB, oral bulge; T, trunk. Scale bars 5 ␮m (G, H), 20 ␮m (A, B, D, F, I), and 30 ␮m (E).
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 91

Dedication: I dedicate this species to Prof. Dr. Aurelio Ser- of three anterior dikinetids followed by 25 monokinetids
rano Delgado, president of the Spanish Microbiology Society with some dikinetids irregularly interspersed. Anterior cil-
and of the VII ECOP-ISOP joint meeting 2015 in Seville, ium of anterior dikinetids possibly lacking or reduced to an
Spain. inconspicuous stump (Figs 8A, G, H, 9E–G; Table 4).
Type material: The holotype slide (reg. no. 843/2015) Oral bulge and head each about 5 × 5 ␮m in vivo, of typ-
and four paratype slides (reg. no. 843/2015–847/2015) with ical Phialina structure, i.e., with very narrowly spaced head
protargol-impregnated specimens have been deposited in the kineties having 10 ␮m long cilia and last kinetid slightly set
Biology Centre of the Upper Austrian Museum in Linz (LI). off; a circumoral kinety difficult to recognize; and a fibrous
The holotype and other relevant specimens have been marked cylinder in head (Figs 8A, D, G–J, 9A, B, E–H; Table 4).
with black ink circles on the coverslip. Occurrence and ecology: As yet found only at type
Description: Phialina serranoi has an ordinary variability locality. Phialina serranoi became rather abundant in the
with most variation coefficients lower than 15% (Table 4), one-week-old sludge sample, suggesting that it prefers
except of macronucleus width (CV 20.3%), the number of microaerobic conditions.
monokinetids in a ciliary row (CV 17.4%), and the number Species comparison: Many Phialina species have been
of dikinetids at begin of the ciliary rows (CV 25.6%). described since the last revision of the genus by Kahl (1930);
Size of extended specimens in vivo 67–100 × 20–30 ␮m, a more recent review is not available. I checked most descrip-
usually about 85 × 27 ␮m, as calculated from some in vivo tions, e.g., Foissner (1983) and Vuxanovici (1959), but all
measurements and the morphometric data in Table 4 adding species have the contractile vacuole in or very near to the
15% preparation shrinkage. Contracts and extends slowly and posterior body end; in some species with acute body end,
contracts only moderately when fixed with Stieve’s solution. the contractile vacuole is slightly subterminal, e.g., in P. min-
Extended body elongate ellipsoid with an average length: ima (Kahl, 1927; redescribed by Foissner et al. 2002) and
width ratio of 4:1 in vivo and of 3.2:1 in protargol preparations in P. vermicularis, as redescribed by Foissner (1983). Thus,
(Figs 8A, G, H, 9E, F; Table 4); unflattened. Most contractile P. serranoi is unique in having the contractile vacuole far
in anterior third of cell; fully contracted cells broadly ellipsoid subterminal, i.e., between mid-body and body end.
or obovate with retracted head (Figs 8K, 9A, B, I). Nuclear
apparatus on average in or slightly posterior to mid-body
(Figs 8A, H, 9A, B, D–F; Table 4). Macronucleus broadly Acknowledgements
ellipsoid or indistinctly reniform, on average 14 × 10 ␮m in
protargol preparations, becomes globular (possibly contracts) Financial support was provided by the Austrian Science
under slight coverslip pressure; of 100 cells investigated, Fund (FWF project P 26325-B16) and the Spanish Micro-
19 have two broadly ellipsoid macronuclear nodules, very biology Society. The technical assistance of Michael Gruber
likely post-conjugates (Foissner and Xu 2007). Micronucleus M.Sc., Dr. Heidi Bartel, and Robert Schörghofer is greatly
attached or near macronucleus, ellipsoid, about 2 × 1 ␮m acknowledged.
in protargol preparations. Contractile vacuole conspicuous
because far subterminal, i.e., between mid-body and poste-
rior end; excretory pore not impregnated with the protargol
References
method used (Figs 8A, 9A, B). Two types of rod-shaped
extrusomes attached to oral bulge and scattered in cytoplasm
Aescht, E., Foissner, W., 1992. Biology of a high-rate activated
(Figs 8A–C, I, 9A, C, D, G, H). Type I about 12 × 0.4 ␮m in sludge plant of a pharmaceutical company. Arch. Hydrobiol.
size, forms bundles in cytoplasm, does not impregnate with Suppl. 90, 207–251.
the protargol method used. Type II extrusomes only 2.5–3 ␮m Al-Shahwani, S.M., Horan, N.J., 1991. The use of protozoa to indi-
long, likely form a ring in oral bulge, impregnate with pro- cate changes in the performance of activated sludge plants. Water
targol. Cortex very flexible, studded with rows of rather Res. 25, 633–638.
refractive granules about 0.8 × 0.4 ␮m in size (Figs 8F, 9D). Ardern, E., Lockett, W.T., 1914. Experiments on the oxidation of
Cytoplasm crammed with food vacuoles 4–7 ␮m across most sewage without the aid of filters. J. Soc. Chem. Ind. 23, 523–539.
containing a corroded starch grain (Figs 8A, 9A), anterior Arévalo, J., Moreno, B., Pérez, J., Gómez, M.A., 2009. Applicabil-
third dark under bright field illumination because studded ity of the Sludge Biotic Index (SBI) for MBR activated sludge
with crystals and globular inclusions 2–5 ␮m in size; both control. J. Hazard. Mater. 167, 784–789.
Arregui, L., Pérez-Uz, B., Zornoza, A., Serrano, S., 2010. A
scattered also throughout body; some cells studded with
new species of the genus Metacystis (Ciliophora, Prostomatida,
about 3 ␮m high, strongly refractive cones, very likely cyst Metacystidae) from a wastewater treatment plant. J. Eukaryot.
spines (Figs 8A, E, 9A–D). Swims fast with head moving Microbiol. 57, 362–368.
to and fro; creeps between sludge flocs like a small worm, Arregui, L., Liébana, R., Calvo, P., Pérez-Uz, B., Salvadó, H., Ser-
showing great flexibility. rano, S., 2013. Bioindication in activated sludge wastewater
Trunk cilia about 8 ␮m long in vivo and in protargol treatment plants. In: Valdez, C.J., Maradona, E.M. (Eds.), Hand-
preparations, arranged in an average of 15 ordinarily spaced, book of Wastewater Treatment. Nova Science Publishers, pp.
meridional or slightly spiral rows composed of an average 277–291.
92 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Banina, N.N., 1983. Peritricha Sessilida in the activated sludge Approach to Sewage Treatment Process: Current Status and Per-
community. Protozoologiya 8, 76–116 (in Russian). spectives. Centro Bazzucchi, Perugia, pp. 93–96.
Barker, A.N., 1942. The seasonal incidence, occurrence and distri- Curds, C.R., 1966. An ecological study of the ciliated protozoa in
bution of protozoa in the bacteria bed process of sewage disposal. activated sludge. Oikos 15, 282–289.
Ann. Appl. Biol. 29, 23–33. Curds, C.R., 1982. The ecology and role of protozoa in aerobic
Barker, A.N., 1943. Biological assay in sewage purification. Proc. sewage treatment processes. Annu. Rev. Microbial. 36, 27–46.
Leeds Phil. Lit. Soc. Sci. Sec. 4, 87–96. Curds, C.R., 1992. Protozoa and the water industry. I–IV. Cambridge
Bayerisches Landesamt für Wasserwirtschaft, 1992. Das University Press, Cambridge, New York, Sydney.
mikroskopische Bild bei der biologischen Abwasserreini- Curds, C.R., Cockburn, A., 1970a. Protozoa in biological sewage-
gung. Informationsberichte des Bayerischen Landesamtes für treatment process – I. A survey of the protozoan fauna of British
Wasserwirtschaft 1/90, 1–101 & Anhang I, II. percolating filters and activated-sludge plants. Water Res. 4,
Bayerisches Landesamt für Wasserwirtschaft, 1999. Das 225–236.
mikroskopische Bild bei der biologischen Abwasserreini- Curds, C.R., Cockburn, A., 1970b. Protozoa in biological sewage-
gung. Informationsberichte des Bayerischen Landesamtes für treatment process – II. Protozoa as indicators in the activated-
Wasserwirtschaft 1/99, 1–166. sludge process. Water Res. 4, 237–249.
Becares, E., Foissner, W., 1994. Redescription of Chilodonatella Curds, C.R., Cockburn, A., Vandyke, J.M., 1968. An experimental
minuta DRAGESCO 1966 (Protozoa, Ciliophora). Linzer Biol. study of the role of the ciliated protozoa in the activated sludge
Beitr. 26, 515–530. process. Water Pollut. Control 67, 312–329.
Bedogni, G., Falanelli, A., Pedrazzi, R., 1991. Evaluation of the De, A., Cybis, L.F., Horan, N.J., 1997. Protozoan and metazoan
abundance ratio between crawling and attached ciliates in the populations in sequencing batch reactors operated for nitrifica-
management of an activated sludge sewage treatment plant. In: tion and/or denitrification. Water Sci. Technol. 35, 81–86.
Madoni, P. (Ed.), Biological Approach to Sewage Treatment Pro- De Marco, N., Gabelli, A., Cattaruzza, C., Petronio, L., 1991. Perfor-
cess: Current Status and Perspectives. Centro Bazzucchi, Perugi, mance of biological sewage treatment plants: some experiences
pp. 229–233. on municipal plants in the province of Pordenone (Italy). In:
Berger, H., Foissner, W., 2003. Biologische Methoden der Madoni, P. (Ed.), Biological Approach to Sewage Treatment
Gewässeranalysen: Ciliaten III-2.1. Illustrated guide and ecolog- Process: Current Status and Perspectives. Centro Bazzucchi,
ical notes to ciliate indicator species (Protozoa, Ciliophora) in Perugia, pp. 247–251.
running waters, lakes, and sewage plants. In: Steinberg, C., Cal- Drzewicki, A., Kulikowska, D., 2011. Limitation of sludge biotic
mano, S., Wilken, K. (Eds.), Handbuch Angewandte Limnologie. index application for control of a wastewater treatment plant
17. Erg. Lfg. , pp. 1–160. working with shock organic and ammonium loadings. Eur. J.
Berger, H., Foissner, W., Kohmann, F., 1997. Bestimmung und Protistol. 47, 287–294.
Ökologie der Mikrosaprobien nach DIN 38410. Gustav Fischer Dubber, D., Gray, N., 2011a. The influence of fundamental design
Verlag, Stuttgart, Jena, Lübeck. parameters on ciliates community structure in Irish activated
Berk, S.G., Gunderson, J.H., 1993. Wastewater Organisms. A Color sludge systems. Eur. J. Protistol. 47, 274–286.
Atlas. Lewis Publishers, Boca Raton, Ann Arbor, London, Dubber, D., Gray, N., 2011b. The effect of anoxia and anaerobia on
Tokyo. ciliate community in biological nutrient removal systems using
Berri, A., Casaschi, R., 1991. Observations of the microfauna and laboratory-scale sequencing batch reactors (SBRs). Water Res.
evaluation of nitrification and denitrification activity of biofilm 45, 2213–2226.
on the RBC. In: Madoni, P. (Ed.), Biological Approach to Sewage Eikelboom, D.H., van Buijsen, H.J.J., 1992. Handbuch für die
Treatment Process: Current Status and Perspectives. Centro Baz- mikroskopische Schlamm-Untersuchung, 3 ed. F. Hirthammer
zucchi, Perugia, pp. 75–81. Verlag, München.
Buck, H., Buck, S., 1980. Mikroorganismen in der Abwasserreini- Eisenmann, H., Letsiou, I., Feuchtinger, A., Beisker, W., Man-
gung. F. Hirthammer Verlag, München. nweiler, E., Hutzler, P., Arnz, P., 2001. Interception of small
Canals, O., Salvadó, H., Auset, M., Hernández, C., Malfeito, J.J., particles by flocculent structures, sessile ciliates, and the basic
2013. Microfauna communities as performance indicators for an layer of a wastewater biofilm. Appl. Environ. Microbiol. 67,
A/O shortcut biological nitrogen removal moving-bed biofilm 4286–4292.
reactor. Water Res. 47, 3141–3150. Fernández-Galiano, D., Guinea, A., Serrano, S., Martín-Cereceda,
Čech, J.S., Hartman, P., Macek, M., 1994. Bacteria and protozoa M., Arregui, L., Rodriguez, B., Campos, I., Calvo, P., Suárez, J.,
population dynamics in biological phosphate removal systems. 1996. Guía práctica de identificación de protozoos ciliados en
Water Sci. Technol. 29, 109–117. estaciones depuradoras de aguas residuales por lodos activos de
Chaudhry, R., Shakoori, A.R., 2012. A new subspecies of a ciliate la comunidad autónoma de Madrid. Private publication by the
Euplotes musicola isolated from industrial effluents. Pak. J. Zool. Dpto. de Microbiología, Facultad de Biología. U.C.M. Comu-
44, 809–822. nidad Autónoma de Madrid. Canal de Isabel II.
Chen, S., Xu, M., Cao, H., Zhu, J., Zhou, K., Xu, J., Yang, X., Foissner, W., 1983. Taxonomische Studien über die Ciliaten
Gan, Y., Liu, W., Zhai, J., Shao, Y., 2004. The activated-sludge des Großglocknergebietes (Hohe Tauern, Österreich) I. Fami-
fauna and performance of five sewage treatment plants in Beijing, lien Holophryidae, Prorodontidae, Plagiocampidae, Colepidae,
China. Eur. J. Protistol. 40, 147–152. Enchelyidae und Lacrymariidae nov. fam. Annln naturh Mus.
Cingolani, L., Cossignani, M., Miliani, R., 1991. The role of micro- Wien 84B, 49–85.
fauna in the prediction and control of the activated sludge Foissner, W., 1993. Colpodea (Ciliophora). Fischer, Stuttgart, Pro-
disfunctions of a municipal plant. In: Madoni, P. (Ed.), Biological tozoenfauna 4, I–X + 798 pp.
 W. Foissner / European Journal of Protistology 55 (2016) 75–94 93

Foissner, W., 2014a. An update of ‘basic light and scanning electron Guggiari, M., Peck, R., 2008. The bacterivorous ciliate Cyclidium
microscopic methods for taxonomic studies of ciliated protozoa’. glaucoma isolated from a sewage treatment plant: molecular
Int. J. Syst. Evol. Microbiol. 64, 271–292. and cytological descriptions for barcoding. Eur. J. Protistol. 44,
Foissner, W., 2014b. Bioindication with protists in the activated 168–180.
sludge process: solution of the taxonomic impediment. In: Helmholtz-Zentrum für Umweltforschung, 2011, June. In Sachen
Sevilla, G.B. (Ed.), Curso sobre Microbiologia Aplicada del Wasser. UFZ-Spezial., pp. 30–31.
Fango Activo. , pp. 425–450. Hu, B., Qi, R., An, W., Xu, M., Zhang, Y., Bai, X., Bao, H., Wen,
Foissner, W., Berger, H., 1996. A user-friendly guide to the ciliates Y., 2013. Dynamics of the microfauna community in a full-
(Protozoa, Ciliophora) commonly used by hydrobiologists as scale municipal wastewater treatment plant experiencing sludge
bioindicators in rivers, lakes, and waste waters, with notes on bulking. Eur. J. Protistol. 49, 491–499.
their ecology. Freshwater Biol. 35, 375–482. Imhoff, K.R., Imhoff, K., 1993. Taschenbuch der Stadten-
Foissner, W., Foissner, I., 1995. Fine structure and systematic twässerung, 28 ed. R. Oldenbourg Verlag, München, Wien.
position of Enchelyomorpha vermicularis (Smith, 1899) Kahl, Jenkins, D., Richard, M.G., Daigger, G.T., 2004. Manual on the
1930, an anaerobic ciliate (Protozoa, Ciliophora) from domestic causes and control of activated slugde bulking, foaming, and
sewage. Acta Protozool. 34, 21–34. other solids separation problems, 3 ed. IWA Publ, London,
Foissner, W., Xu, K., 2007. Monograph of the Spathidiida UK.
(Ciliophora, Haptoria). Volume I: Protospathidiidae, Arcu- Kahl, A., 1926. Neue und wenig bekannte Formen der
ospathidiidae, Apertospathulidae. Monogr. Biol. 81, 1–485. holotrichen und heterotrichen Ciliaten. Arch. Protistenk. 55,
Foissner, W., Skogstad, A., Pratt, J.R., 1988. Morphology and infra- 197–438.
ciliature of Trochiliopsis australis n. sp., Pelagohalteria viridis Kahl, A., 1927. Neue und ergänzende Beobachtungen holotricher
(Fromentel, 1876) n. g., n. comb., and Strobilidium lacustris n. Ciliaten I. Arch. Protistenk. 60, 34–129.
sp. (Protozoa, Ciliophora). J. Protozool. 35, 489–497. Kahl, A., 1930. Urtiere oder Protozoa I: Wimpertiere oder Ciliata
Foissner, W., Blatterer, H., Berger, H., Kohmann, F., 1991. (Infusoria) 1. Allgemeiner Teil und Prostomata. Tierwelt Dtl. 18,
Taxonomische und ökologische Revision der Ciliaten des Sapro- 1–180.
biensystems – Band I: Cyrtophorida, Oligotrichida, Hypotrichia, Kinner, N.E., 1984. An evaluation of the feasibility of using pro-
Colpodea. Informationsberichte des Bayerischen Landesamtes tozoa and metazoa as indicators of RBC effluent quality. In:
für Wasserwirtschaft 1/91., pp. 1–478. Second International Conference on Fixed-film Biologial Pro-
Foissner, W., Berger, H., Kohmann, F., 1992. Taxonomische und cesses, 10–12 July, Arlington, Virginia, pp. 74–122.
ökologische Revision der Ciliaten des Saprobiensystems – Band Kunst, S., Helmer, C., Knoop, S., 2000. Betriebsprobleme auf
II: Peritrichia, Heterotrichida, Odontostomatida. Informations- Kläranlagen durch Blähschlamm, Schwimmschlamm, Schaum.
berichte des Bayerischen Landesamtes für Wasserwirtschaft Handbuch zur Identifizierung fädiger Bakterien. Springer,
5/92., pp. 1–502. Berlin, Heidelberg, New York, pp. 175 pp.
Foissner, W., Berger, H., Kohmann, F., 1994. Taxonomische und Lee, S., Basu, S., Tyler, C.W., Wei, I.W., 2004. Ciliate populations
ökologische Revision der Ciliaten des Saprobiensystems – Band as bio-indicators at Deer Island Treatment Plant. Adv. Environ.
III: Hymenostomata, Prostomatida, Nassulida. Informations- Res. 8, 371–378.
berichte des Bayerischen Landesamtes für Wasserwirtschaft Lemmer, H., Lind, G., 2000. Blähschlamm, Schaum, Schwimm-
1/94., pp. 1–548. schlamm. Mikrobiologie und Gegenmaßnahmen. F. Hirtham-
Foissner, W., Berger, H., Blatterer, H., Kohmann, F., 1995. mer, München, pp. 176 pp.
Taxonomische und ökologische Revision der Ciliaten des Sapro- Liebmann, H., 1936. Die Ciliatenfauna der Emscherbrunnen. Z.
biensystems – Band IV: Gymnostomatea, Loxodes, Suctoria. Hyg. InfektKrankh. 118, 555–573.
Informationsberichte des Bayerischen Landesamtes für Wasser- Liebmann, H., 1951. Handbuch der Frischwasser- und Abwasser-
wirtschaft 1/95., pp. 1–540. biologie. Biologie des Trinkwassers, Badewassers, Fis-
Foissner, W., Berger, H., Schaumburg, J., 1999. Identification and chwassers, Vorfluters und Abwassers, vol. I. R. Oldenbourg,
ecology of limnetic plankton ciliates. Informationsberichte des München.
Bayerischen Landesamtes für Wasserwirtschaft 3/99., pp. 1–793. Liebmann, H., 1958. Handbuch der Frischwasser- und Abwasser-
Foissner, W., Agatha, S., Berger, H., 2002. Soil ciliates (Protozoa, biologie. Biologie des Trinkwassers, Badewassers, Fis-
Ciliophora) from Namibia (Southwest Africa), with emphasis on chwassers, Vorfluters und Abwassers, vol. II. R. Oldenbourg,
two contrasting environments, the Etosha region and the Namib München.
Desert. Denisia 5, 1–1459. Luna-Pabello, V.M., Plisson-Saune, S., Paul, E., Duran de Bazúa,
Ganner, B., Unterweger, A., Jäger, P., 2002. Die Biologie der C., 1996. Ciliatological characterization of a biological reactor
Salzburger Kläranlagen im Zeitraum von 1991 bis 2000. that eliminates nitrogen with intermitent aeration. Rev. lat.-amer.
Zur Evaluation der Beurteilungskriterien bei der mikroskopis- Microbiol. 38, 89–96.
chen Belebtschlammuntersuchung kommunaler Kläranlagen mit Lynn, D.H., 2008. The Ciliated Protozoa: Characterization, Classi-
Stickstoffelimination. Amt der Salzburger Landesregierung. fication and Guide to the Literature, 3 ed. Springer, Dordrecht.
Gewässerschutz 6, 115–170. Madoni, P., 1991. Biological Approach to Sewage Treatment Pro-
Gori, F., Greco, M., Guarnieri, G., Minelli, L., 1991. Microfauna cess: Current Status and Perspectives. Centro Luigi Bazzucchi,
analysis in performance evaluation of the activated sludge depu- Perugia.
ration plant in Foligno (Italy). In: Madoni, P. (Ed.), Biological Madoni, P., 1994. A sludge biotic index (SBI) for the evaluation of
Approach to Sewage Treatment Process: Current Status and Per- the biological performance of activated sludge plants based on
spectives. Centro Bazzucchi, Perugia, pp. 97–99. the microfauna analysis. Water Res. 28, 67–75.
94 W. Foissner / European Journal of Protistology 55 (2016) 75–94

Marsh, T.L., Liu, W.-T., Forney, L.J., Cheng, H., 1998. Beginning Pérez-Uz, B., Arregui, I., Calvo, P., Salvadó, H., Fernández, N.,
a molecular analysis of the eukaryal community in activated Rodríguez, E., Zornoza, A., Serrano, S., 2010. Assesment of
sludge. Water Sci. Technol. 37, 455–460. advanced wastewater treatments for nitrogen removal searching
Martín-Cereceda, M., Serrano, S., Guinea, A., 1996. A compara- for plausible efficiency bioindicators. Water Res. 44, 5059–5069.
tive study of ciliated protozoa communities in activated-sludge Rehman, A., Shakoori, F.R., Shakoori, A.R., 2006. Heavy metal
plants. FEMS Microbiol. Ecol. 21, 267–276. resistant ciliate, Euplotes mutabilis, isolated from industrial
Martín-Cereceda, M., Serrano, S., Guinea, A., 2001. Biofilm com- effluents can decontaminate wastewater of heavy metals. Bull.
munities and operational monitoring of a rotating biological Environ. Contam. Toxicol. 76, 907–913.
contractor system. Water Air Soil Pollut. 126, 193–206. Salvadó, H., Mas, M., Menéndez, S., Gracia, M.P., 2001. Effects
Martin-Cereceda, M., Guinea, A., Bonaccorso, E., Dyal, P., of shock loads of salt on protozoan communities of activated
Novarino, G., Foissner, W., 2007. Classification of the peritrich sludge. Acta Protozool. 40, 177–185.
ciliate Opisthonecta matiensis (Martin-Cereceda et al. 1999) as Santos dos Araújo, L., Ferreira, V., Pereira, M.O., Nicolau, A., 2014.
Telotrochidium matiense nov. comb., based on new observations Relationship between protozoan and metazoan communities and
and SSU rDNA phylogeny. Eur. J. Protistol. 43, 265–279. operation and performance parameters in a textile sewage acti-
Matsunaga, K., Kubota, K., Harada, H., 2014. Molecular diversity vated sludge system. Eur. J. Protistol. 50, 319–328.
of eukaryotes in municipal wastewater treatment processes as Scherb, K., 1968. Zur Biologie des belebten Schlammes. Münchn.
revealed by 18S rRNA gene analysis. Microbes Environ. 29, Beitr. Abwass.- Fisch.- Flussbiol. 5, 158–205.
401–407. Serrano, S., Arregui, L., Perez-Uz, B., Calvo, P., Guinea, A., 2008.
Morishita, I., 1970. Studies on protozoa-populations in activated Guidelines for the Identification of Ciliates in Wastewater Treat-
sludge of sewage and waste treatment plants. Jpn. J. Protozool. ment Plants. IWA Publishing, UK.
3, 1–13. Shizheng, W., Zhengxue, M., 1995. Propyxidium elongatum n. sp.
Mudrak, K., Kunst, S., 1994. Biologie der Abwasserreinigung, 4 ed. the ciliate of genus Propyxidium from industrial wastewater of
Gustav Fischer Verlag, Stuttgart. Yinchuan, China. In: Asai, H., Naitoh, Y. (Eds.), Proceedings of
Oberschmidleitner, R., Aescht, E., 1996. Taxonomische Unter- the 4th Asian Conference on Ciliate Biology and the Interna-
suchungen über einige Ciliaten (Ciliophora, Protozoa) aus tional Symposium on Cell Motility and Cytogenesis. Tokyo, pp.
Belebtschlämmen oberösterreichischer Kläranlagen. Beitr. 234–236.
Naturk. Oberösterr. 4, 3–30. Silva da, S.B.A., Silva-Neto da, I.D., 2001. Morfologia dos pro-
Paiva, T. da S., Silva-Neto, I.D. da, 2004. Description of Parento- tozoários ciliados presentes em um reator experimental de
cirrus brasiliensis sp. n. (Ciliophora: Spirotrichea), a new ciliate tratamento de esgoto por processo de lodos ativados. Rev. bras.
protist present in activated sludge. Zootaxa 504, 1–10. Zoociências Juiz de Fora 3, 203–229.
Paiva, T. da S., Silva-Neto, I.D. da, 2005. Deviata estevesi sp. Siqueira-Castro, I.C.V., Paiva da, T.S., Silva-Neto da, I.D., 2009.
n. (Ciliophora: Spirotrichea), a new ciliate protist from a Morphology of Parastrongylidium estevesi comb. nov. and Devi-
restinga lagoon in Rio de Janeiro, Brazil. Acta Protozool. 44, ata brasiliensis sp. nov. (Ciliophora: Stichotricha) from a sewage
351–362. treatment plant in Rio de Janeiro, Brazil. Zoologia 26, 774–786.
Pauli, W., Jax, K., Berger, S., 2001. Protozoa in wastewater Sládeček, V., 1973. System of water quality from the biological
treatment: function and importance. In: Beek, B. (Ed.), The point of view. Arch. Hydrobiol. Beih. Ergebn. Limnol. 7, 1–218,
Handbook of Environmental Chemistry, Vol. 2, Part K: Biodegra- I–IV.
dation and Persistence. Springer, Berlin and Heidelberg, pp. Toman, M.J., 2002. Biological assessment of wastewater treatment
203–252. plant conditions using the sludge biotic index. Verh. Internat.
Pedrazzani, R., 2014. Activated sludge monitoring in a real scale Verein. Limnol. 28, 692–694.
MBR (membrane bioreactor) plant. In: Grupo Bioindicación Uhlmann, D., 1982. Hydrobiologie. Ein Grundriß für Ingenieure
Sevilla (Ed.), Curso sobre Microbiologia Aplicada del Fango und Naturwissenschaftler, 2 ed. G. Fischer Verlag, Stuttgart.
Activo. , pp. 389–397. Vuxanovici, A., 1959. Contributii la studiul unor infuzori holotrichi.
Penard, E., 1922. Études sur les Infusoires d’Eau Douce. Georg and Studii Cerc. Biol. (Biol. Anim.) 11, 307–335 (in Roumanian with
Cie, Genève. Russian and French summary).
Pérez-Uz, B., Franco, C., Martín-Cereceda, M., Arregui, L., Cam- Weisse, T., 2014. Ciliates and rare biosphere – community ecology
pos, I., Serrano, S., Guinea, A., Fernández-Galiano, D., 1998. and population dynamics. J. Eukaryot. Microbiol. 61, 419–433.
Biofilm characterization of several wastewater treatment plants Zhou, K., Xu, M., Dai, J., Cao, H., 2006. The microfauna commu-
with rotating biological contactors in Madrid (Spain). Water Sci. nities and operational monitoring of an activated sludge plant in
Technol. 37, 215–218. China. Eur. J. Protistol. 42, 291–295.