Pathogen densities
in nature
Bacteria in aquaristic systems
Numerous species of bacteria,
viruses and fungi live in aquatic
systems. Bacteria and algae
are the numerically dominating
organisms of inshore waters and
form a great part of the biomass.
Their metabolism affects and con-
trols the flow of energy in aquatic
habitats and of the nutrient cycles
and processes of energy conver-
sion happening within.
Bacteria, small organisms without
a nucleus, inhabit practically all
habitats of the earth. In principle
Der Makropode – Volume 30 – 1 / 2008 21
By Gerhard Ott
there is no difference between
those bacteria that inhabit the
soil and those that live in waters.
Within a water body many spe-
cies can occur planktonically, that
means floating, as well as ses-
sile, which means on substrates.
Above that, there are forms that
predominantly live sessile and
others that predominantly live
freely floating. In streams and
stagnant waters there are corre-
spondingly different populations
of bacteria in nature.
The aquaristic role of bacteria is
emphasized repeatedly if it comes
and the aquarium
to filter bacteria. In most cases
these are sessile bacteria. Hence,
a large surface of filter substrates
is aspired. But also in the aquaria
themselves those bacteria can be
found, for example in appropriate
substrates. In the optimal case,
these bacteria are among others
Nictrobacter of the nitrogen cycle
which should be obtained and
cultivated. Therefore, the expe-
rienced aquarist never rinses out
the filtration medium completely.
In order to always retain a frac-
tion of the bacterial colonies, the
rinsing water should not be to hot

Filtration, a method for the reduction of organic loading in the aquarium
water
or to cold. Thus, the experienced
aquarist starts a new aquarium
with the filter mud from a matured
aquarium or uses an according
substance offered in pet shops.
Bremer (1995, 1996a, b) and Kasse-
beer (1996, 1999) pronounced
repeatedly the important role of
floating bacteria in the aquarium
and in fish breeding facilities.
Non-sessile bacteria are at the
same time indicators for organic
22 Der Makropode – Volume 30 – 1 / 2008
loading of the water. Saprophagic
living bacteria are those that feed
from decaying and fouling matter
meaning organic waste. In hydrol-
ogy, especially when dealing with
rivers, they are a measure for the
organic load of water bodies (sap-
robe system). Saprophytic bacte-
ria are characterized in that way
that they are capable of exploit-
ing mainly proteins and simple
carbohydrates. Pure water sap-
rophages are able to exploit even
very low concentrations of nutri-
ents (Wright & Hobbie 1966).
Further Saprophages (decompo-
sition exploiters)
Fungi are even more variegated
then bacteria. They exhibit a real
nucleus and can be single- or mul-
ticellular. In offshore waters they
occur in high species numbers. An
assignment between terrestrial
and aquatic species or groups is
not easy. Biologically interested
aquarists are familiar with genus
names like Achlya, Aphanomyces,
Branchiomyces and Saprolegnia.
They are Phycomyceta that can
be understood as actual aquatic
fungi. Apart from simple struc-
tured proteins, carbohydrates and
lipids they can also exploit pec-
tins, cellulose type matter, lignin
and chitin. Aquatic Deuteromyces
growing on decaying litter can
exploit fibers and play an impor-
tant role in aquatic biotopes as
well.
Bacteria, fungi and fishes
Between bacteria and fishes exist
manifold interactions. These
interactions express themselves
amongst others as attacks of
the microorganisms against the
organism of the fish. Especially
the breeder of aquarium fishes
knows that within breeding tanks
with high stocking densities and
high feeding rates, a periodic and
extensive partial water exchange
has to be carried out to keep the
fish brood healthy. It is also impor-
tant to consider the microbiologi-
cal situation in the aquarium apart
from the filter.
The optimum of many bacteria
is within the neutral to slightly
alkaline range. Generally, a pH
below 7 acts inhibitory on most
bacteria concerning growth and
reproduction. Many fungal spe-
cies can indeed exist in slightly
to very acid waters. The aquar-
ist is aware of the so called soft
water from the natural habitat of
many aquarium fishes. Soft water
means, that it is low in carbonate;
most of the times it is also poor in
other ions, so that the electric con-
ductibility (“conductance”) is low
to very low. Basically, calcium car-
bonate and magnesium carbon-
ate are meant with that. From the
aquaristic and fish breeding point
of view, amongst others Beyer
(1998) noted the importance of
calcium ions Ca²+. Soft waters are
often very low in bacteria because
bacteria need a certain concentra-
tion of ions for growing.
Mycoses (fungal infections) in
fishes appear only periodically if
the outer skin is affected. Fungal
spores can not germinate on a
healthy glycocalyx (slime coat)
and the epidermis below. If the
glycocalyx is affected by bacterial
infections in advance fungi can
grow secondarily.
Germs? Number of germs? Ger-
minal density?
These perceptions are a wonderful
example on how scientific appear-
ing terms are sometimes used in
an unreflected way. The Germanic
origin of the word “germ” is
known in botany and greeneries
as offshoot of herbal organisms,
meaning that what grows out
of a grain. But the term “germ”
has also a second meaning in the
sense of “germ” = pathogen.
In hygienics the term bacterial
count is mostly used. In prin-
ciple that is incorrect. Because it
is about the number of bacteria
(“germs”) in a defined volume,
the correct termination would
be germ density. I do not want
to miss to mention that I owe
this information Dr. Stefan Hetz,
a physiologist at the Humboldt
University of Berlin (Similarly, in
aquaristics the term conductivity
number (note by translator: this
is a direct translation which does
not seem to exist in the English
language) is common although
electric conductivity is meant).
Bacterial densities are measured
Der Makropode – Volume 30 – 1 / 2008 23
If a filter inlet produces foam the organic load is very high already
in CFU, colony forming units per
milliliter. 104 CFU/ml imply that in
one milliliter (= 1 cubic centime-
ter) water are 10.000 bacteria.
According to the drinking water
ordinance in Germany bacte-
ria like Salmonella (typhus and
paratyphoid), Shigella (dysen-
tery), Vibrio cholerae (cholera),
Pseudomonas (among others
pneumonia) and Legionella (legio-
nellosis) have to be undetectable
in drinking water. Bioindicators
are coliform bacteria like Escheri­
chia coli. Here, the benchmark is
10² CFU/ml, after completion of
the preparation it is 20 CFU/ml
(see Mutschmann & Stimmelmayer
1995, p. 155 ff.).
Assessing the bacterial density –
estimating loading of the water
For natural and, above that, fish-
ery-biological interesting water
bodies and aquaculture facilities
specifying the bacterial density is
a common method to quantify the
loading of the water. In aquaristics
these methods are used for some
years now. Using adequate tests –
coming from the medical hygiene
– the biologically effective bacte-
rial densities can be determined
by aquarists. The actual number
of bacteria in the water is normally
higher as the unit CFU indicates
(Rheinheimer 1991) and in fact it
is about an order of magnitude.
The handy methods for measure-
ment only measure those bacteria
which establish colonies.
A plate is wetted with a defined
amount of water, or a paddle
with bacterial culture substrate is
dipped, incubated and afterwards
one can count the amount of colo-
nies and evaluate it according to a
scaling. The determined numbers
of bacteria are the overall results
from those factors that promote
or impede bacterial development.
In high organic loading (food), a
temperature above 20 °C, suffi-
cient oxygen supply, neutral and
alkaline pH values and a reduc-
tion potential below 200 mV,
saprophytic bacteria develop opti-
mally. Tanning agents (tannines,
catechine, fulvo acids and humic
acids) inhibit their growth. The
bacterial density in open water
is a measure for the organic pol-
lution of water bodies. In nature
as well as in artificial ecosystems
like aquaria.
Biotest HYCON Keimindikatoren
Dip Slides
For aquaristic purposes the deter-
mination of the bacterial density
can be accomplished with the

Colony forming units (CFU) per milliliter
Biotest HYCON Keimindikatoren
Dip Slides. The culture medium
plate is dipped in the water and
incubated for 24 to 48 hours at 30
°C in the included tube. The light
yellow colored medium GK-A dis-
plays the amount of bacteria in
CFU/ml = colony forming units
per milliliter. The brown reddish
medium displays on the backside
a selective verification of coliform
bacteria also in CFU/ml, which
simply means: the degree of con-
tamination with fecal bacteria.
10² CFU/ml water with very
low bacteria count; in an aquar-
ium this is almost trade – if not in stock in a phar-
unfeasible without devices like
ozonization or UV radiation
10³ CFU/ml water with low
bacteria count
104 CFU/ml typically found in
heavily stocked community tanks
and are tolerated by many fishes
105 CFU/ml harms fish
Lately, these tests are also sold
in aquaristic pet shops. So far the
interested aquarist had to rely on
specialized laboratory trade. One
supplier for these test kits is the
company Windaus Labortechnik,
Bauhofstrasse 9, 38678 Clausthal-
Zellerfeld, Fax 053 23/ 4 01 96,
http://www.windaus.de, which
24 Der Makropode – Volume 30 – 1 / 2008
delivers these measuring sets
starting at 5 pieces.
The method can also be used in
the field when conducting habi-
tat studies for aquaristic and lim-
nological purposes (Ott 2000a,
b). This way one can for example
easily determine if habitats have
been affected by anthropogenic
influences: Mostly, close to vil-
lages higher densities of coliform
bacteria are detectable. One rule
of experienced wanderers and
trekkers is to always bathe or pull
water above a settlement. One
knows why...
Cult-Dip combi
With the Biotest HYCON Keimind-
ikatoren Dip Slides total bacterial
densities in CFU and coliform bac-
teria (also in CFU) can be deter-
mined. The Cult-Dip Combi by
Merck KgaA (64271 Darmstadt)
consists of dipping culture media
for determination of bacterial
densities, yeasts and mold. The
media are covered with a pallid
TTC agar and orange colored
potato glucose agar. The culture
substrates have to be in contact
for 5 to 10 s with the fluid to be
tested, i.e. aquarium water. Both
agar sides of the nutrient medium
must be wetted completely. Sur-
plus water should drip off. The
tubes are incubated upright at 27
to 30 °C. In the lab this is done in
an incubator. The aquarist should
be able to find ideal conditions
above a light housing. In the field
in tropic regions these conditions
correspond with the surrounding
temperatures. After incubation
for 24 to 48 hours the result can
be read from the TTC agar. Yeasts
and filamentous fungi grow on the
potato glucose agar after incuba-
tion for approximately 72 hours.
If the incubation is conducted at
room temperature, the results
can be interpreted after 48 to 96
hours (that is 4 to 7 days). After
the incubation, the colony densi-
ties of the cultures can be com-
pared to the included samples.
The majority of bacteria grows as
red colonies. If colorless colonies
occur, they should be accounted
in the determination of CFU, too.
It always depends on the density
of the colonies, not on the size of a
single colony. On the “fungi side”
may be growing mere filamentous
fungi, pure yeasts or an admixture
of yeasts and filamentous fungi.
Filamentous fungi form wool
like colonies and yeasts round,
humpy and dull colonies which
sometimes have extensions. For a
microscopic clarification colonies
can be collected from the culture
medium. – These measurement
sets in the laboratory specialized
macy – normally much cheaper
than in a pet shop.
Typical aquaristic results from
measurements
In principle low bacterial densities
are desired in aquaria. Aquarium
water of which sampling resulted
in high bacterial densities is not
only rich of saprophytic bacteria
in the open water but also organi-
cally polluted. Aquarium water
with low bacterial counts of 10³
CFU/ml is intended. Typically, in
community tanks 104 CFU/ml and
more are found. In sales tanks in
shops values above 106 CFU/ml
have been recorded. In breed-
ing tanks the bacterial densities
should be reduced with appropri-
ate methods because larvae and
young fishes do not have a fully
developed immune system yet.
Which consequences does this
have for aquaristics and the keep-
ing of fishes in aquaria?
In principle it can be stated: In
aquaristic hydrology there is an
array of parameters. These are
among others pH value, acid bind-
ing capacity, degree of hardness,
temperature as well as water
ingredients of favored and non-
favored nature. This is not only
about applying a new indicator. It
is rather about understanding the
coherences. A great importance is
connected to the pH value of the
water when it comes to keeping
fishes. Here, it is not necessar-
ily the concentration of hydrogen
ions the criterion but the impact of
the pH value. A comparative mea-
Der Makropode – Volume 30 – 1 / 2008 25
surement of bacterial densities
shows: Low pH values usually
also indicate a low bacterial load-
ing. With increasing pH values the
bacterial loading accelerates as
well. The slime coating of fishes is
occupied with fending the increas-
ing attacks of bacteria. More than
once it leads to secondary infec-
tions, e.g. Oodinium, because the
slime coating of fishes is preaf-
fected due to defenses against
bacteria.
In typical black waters of the trop-
ics barely 10² up to 10³ CFU/ml are
found. In a meso saprobe central-
European lake and aquarium it
absolutely can be up to 106 CFU
and more. Typical black water
fishes are, with their epidermal
skin and above lying slime coating,
are not adapted to high bacterial
densities and are affected easily
by pathogens, if these are present
in high rates. Here, we know only
little on the actual relationships in
nature and specific adaptations
of certain species (Ott 2000 a, b).
Some labyrinth fishes derive from
such water bodies. Therefore
keeping bacterial densities low
is the goal of every responsible
aquarist of labyrinth fishes. The
available methods for the determi-
nation of bacterial densities in the
aquarium water can amend the
conventional methods of water
quality characterization.
The consequences resulting from
these conclusions and methods
to reduce bacterial densities in
aquaria are known in aquaris-
tic practice: carbon dioxide (or
other “acidification” methods),
peat or peat extract (Wendenburg
1999), alder catkins and accord-
ing extracts (Dengler 2002, Gut-
jahr 1998, Steinle 1998). Some also
take tea. Planting of rapidly grow-
ing water plants (Horst 1999), in
breeding practices with meth-
ylene blue, sodium chloride and
other chemicals. Water changes,
UV radiation, ozonization and
microfilters (with diatoms or car-
bonite as substrate) and large-
area filters (“Hamburger Filter”)
are further methods.
For quite some time the decora-
tion of landscape aquaria with
leaves is a common procedure in
aquaristics. It has been pushed
back a little by the modern and
partly sort of clinically clean
aquaria. Still it can be encountered
at experienced aquarists. This is
because leaves are not only deco-
ration but also fulfill some biologi-
cal functions: For many fishes it
serves as natural cover and pro-
tection, sometimes as spawn-
ing ground. Several substances
that are released to the water by
leaves act positively on biologi-
cal balancing processes within
the ‘mini habitat’ aquarium. For
some fishes leaves even serve as
food or food complement. There
are also according supplements
and extracts, even artificial humic
substance preparations (Geisler
1999, Kühnert 1981, Plöger 2001,
Ott 2003a).
Mostly, leaves of oaks (Quercus) or
beeches (Fagus) are – because of
their ingredients – used. Recently,
Catappa leaves or preparations
herefrom with antiseptic proper-
ties are advertised by aquaristic
commerce: “Due to the extensive
decrease of infective pressure in
the aquarium water, the leaves of
the Indian almond act positively
on the transfer of sensible fishes.
Especially successful are the
leaves applied on imported fish
like discus fishes, neon fishes and
clown loaches,” it is advertised
inter alia (see also Ott 2003b).
The typical black water labyrinth
fishes which are mostly kept by
members of the IGL are appar-
ently not an commercially appeal-
ing target group.
Summary
The method of germ determina-
tion via CFU in aquarium water
can amend the common aquaristic
methods of water quality charac-
terization. The method can also be
26 Der Makropode – Volume 30 – 1 / 2008
used in the field of habitat stud-
ies for aquaristic and limnological
purposes. Above that, the aquar-
ist himself can test if commer-
cially offered substances act in
the according bacteria inhibiting
way.
The investigation and the under-
standing of ecological effect
coherences confirms the prag-
matical experience in aquaristics
and enhance the knowledge for
the living needs of fishes. It also
becomes clear that the “decora-
tion” within an aquarium is not
only “ornamentation” but beyond:
environmental enrichment from
the ethological and even agent
from an ecological point of view.
The question of effectiveness
which is outlined by headwords
like pH value, filter efficiency,
additives to the aquarium water,
conditioners and bacterial count
in the open water can contribute
to the competence of aquarists
when it is causally better per-
ceived.
Literature
Beyer, P. 1998. Das Calcium und
die „Weichwasserfische“. 3. VDA-
Süßwasser-Symposium 7.–8. Nov.
in Fulda, 47–55.
Bremer, H. 1995. Bakterien im
Aquarium. Ihre Rolle als Indika-
tor und Wirkfaktor. DATZ 48(4):
257–259.
Bremer, H. 1996a. Bakterien in
Brut- und Vorstreckanlagen –
Indikator, Wirkung und Methode.
Fischer & Teichwirt (8): 314–316.
Bremer, H. 1996b. Bakterien im
Süßwasser – Bedingungen für
Leben und Tod. Symposiumsband
des 2. VDA-Süßwassersymposi-
ums vom 26.–27.10.1996 in Fulda,
73–78.
Dengler, A.T. 2002. Die Erle – eine
Bereicherung für die Aquaristik.
Vereinsjournal Aquarienfreunde
Dachau, 38–39.
Geisler, R. 1999. Huminstoffe in
der Aquaristik? Vortrag beim 25.
Symposion der Aquaristischen
Interessengemeinschaft am 29.
Mai 1999 in Bielefeld.
Gutjahr, A. 1998. Sind Erlenzäpf-
chen in Vergessenheit geraten?
TI Aquaristik Fachmagazin 30(5):
72.
Horst, K. 1999. Aquarienpflanzen
sind die beste Fisch- und Was-
serpflege. Aquarium Heute 17(2):
325–328.
Kassebeer, G. 1996. Die Bedeutung
der Bakterienflora für die Funktion
des Aquariums. Symposiumsband
des 2. VDA-Süßwassersymposi-
ums vom 26.–27.10.1996 in Fulda,
63–73.
Kassebeer, G. 1999. Hat die Keim-
zahl eine aquaristische Bedeu-
tung? Vortrag beim 25. Symposion
der Aquaristischen Interessenge-
meinschaft am 29. Mai 1999 in
Bielefeld.
Kühnert, M. 1981. Biochemische
und pharmakologisch toxikologi-
sche Eigenschaften von isolierte
hergestellten Huminsäuren. IMIG
Symp. „Torf in der Medizin“ 3:
22–42.
Mutschmann, J. & Stimmelmayr, F.
1995. Taschenbuch der Wasser-
versorgung. Stuttgart, 11. Aufl.
Ott, G. 2000. Keimzahluntersu-
chungen an indonesischen Süßge-
wässern und ihre aquaristischen
Konsequenzen. Vortrag beim
VDA-Symposion am 25. Novem-
ber in Fulda.
Ott, G. 2001. Haben Keimzahlen
eine aquaristische Bedeutung? 1.
Teil: Problemstellung, Messme-
thoden und Befunde. Aquarium
heute 19(1): 43–45. 2. Teil: Steue-
rung der bakteriellen Belastung
im Aquarium. Aquarium heute
19(2): 48–50.
Ott, G. 2002. Warum verpilzen
Fischeier? 1. Teil: Biologische
Tatsachen und Zusammenhänge.
Aquaristik Fachmagazin 34(164):
43–45. 2. Teil: Folgerungen für die
aquaristische Praxis. Aquaristik
Fachmagazin 34(165): 52–55.
Ott, G. 2003a. Für Sie getestet –
Dupla-Cur, Tonikum oder Kosme-
tik? Das Aquarium 37(5): 71–75.
Ott, G. 2003b. Für Sie getestet –
Seemandelbaumblätter im Aqua-
rium. Das Aquarium 37(7): 18–21.
Plöger, J. 2001. Ein synthetischer
Huminstoff. Aquaristik Fachma-
gazin 33(6): 115.
DER
ISSN 0937-177X

MAKROPODE
1/08
30. Jahrgang – April 2008
Zeitschrift der IGL
Internationale Gemeinschaft für Labyrinthfische

Der Makropode – Volume 30 – 1 / 2008 1