Nitrifying Bacteria Facts

Fritz Industries
One of the most important, and least understood, aspects of successful aquarium keeping is biological
filtration and its function in the nitrogen cycle. Traditionally, novice aquarists become disillusioned at
the frequently experienced high death rates of their aquatic pets after setting up a new aquarium.
Statistically, as much as 60% of the fish sold for a new aquarium will die within the first 30 days. 2 out
of every 3 new aquarists abandon the hobby within the first year.
Known as "New Tank Syndrome" these fish are poisoned by high levels of ammonia (NH 3)
that is produced by the bacterial mineralization of fish wastes, excess food, and the
decomposition of animal and plant tissues. Additional ammonia is excreted directly into the
water by the fish themselves. The effects of ammonia poisoning in fish are well documented.
These effects include: extensive damage to tissues, especially the gills and kidney;
physiological imbalances; impaired growth; decreased resistance to disease, and; death.
Nitrite poisoning inhibits the uptake of oxygen by red blood cells. Known as brown blood
disease, or methemoglobinemia, the hemoglobin in red blood cells is converted to
methemoglobin. This problem is much more severe in fresh water fish than in marine
organisms. The presence of chloride ions (CL-) appears to inhibit the accumulation of nitrite in
the blood stream.
The successful aquarist realizes the importance of establishing the nitrogen cycle quickly and with
minimal stress on the aquariums inhabitants. Aquarium filtration has advanced from the old box filters
filled with charcoal and glass wool to undergravel filters, then trickle filters, and most recently fluidized bed filters. Every advance has been to improve upon the effectiveness of biological filtration
which in turn increases the efficiency of the nitrogen cycle. The availability of advanced high-tech
filtration systems has lent added importance to the understanding of basic aquatic chemistry.
Nitrifying bacteria are classified as obligate chemolithotrophs. This simply means that they
must use inorganic salts as an energy source and generally cannot utilize organic materials.
They must oxidize ammonia and nitrites for their energy needs and fix inorganic carbon
dioxide (CO2) to fulfill their carbon requirements. They are largely non-motile and must
colonize a surface (gravel, sand, synthetic biomedia, etc.) for optimum growth. They secrete a
sticky slime matrix which they use to attach themselves.
Species of Nitrosomonas and Nitrobacter are gram negative, mostly rod-shaped, microbes ranging
between 0.6-4.0 microns in length. They are obligate aerobes and cannot multiply or convert ammonia
or nitrites in the absence of oxygen.

Nitrifying bacteria have long generation times due to the low energy yield from their oxidation
reactions. Since little energy is produced from these reactions they have evolved to become
extremely efficient at converting ammonia and nitrite. Scientific studies have shown that
Nitrosomonas bacterium are so efficient that a single cell can convert ammonia at a rate that
would require up to one million heterotrophs to accomplish. Most of their energy production
(80%) is devoted to fixing CO2 via the Calvin cycle and little energy remains for growth and
reproduction. As a consequence, they have a very slow reproductive rate.
Nitrifying bacteria reproduce by binary division. Under optimal conditions, Nitrosomonas may double
every 7 hours and Nitrobacter every 13 hours. More realistically, they will double every 15-20 hours.
This is an extremely long time considering that heterotrophic bacteria can double in as short a time as
20 minutes. In the time that it takes a single Nitrosomonas cell to double in population, a single E. Coli
bacterium would have produced a population exceeding 35 trillion cells.
None of the Nitrobacteraceae are able to form spores. They have a complex cytomembrane (cell wall)
that is surrounded by a slime matrix. All species have limited tolerance ranges and are individually
sensitive to pH, dissolved oxygen levels, salt, temperature, and inhibitory chemicals. Unlike species of
heterotrophic bacteria, they cannot survive any drying process without killing the organism. In water,
they can survive short periods of adverse conditions by utilizing stored materials within the cell. When
these materials are depleted, the bacteria die.

Biological Data
There are several species of Nitrosomonas and Nitrobacter bacteria and many strains among
those species. Most of this information can be applied to species of Nitrosomonas and
Nitrobacter in general., however, each strain may have specific tolerances to environmental
factors and nutriment preferences not shared by other, very closely related, strains. The
information presented here applies specifically to those strains being cultivated by Fritz
Industries, Inc.
Temperature
The temperature for optimum growth of nitrifying bacteria is between 77-86 F (25-30 C).
Growth rate is decreased by 50% at 64 F (18 C).
Growth rate is decreased by 75% at 46-50 F.
No activity will occur at 39 F (4 C)
Nitrifying bacteria will die at 32 F (0 C).
Nitrifying bacteria will die at 120 F (49 C)
Nitrobacter is less tolerant of low temperatures than Nitrosomonas. In cold water systems, care must be
taken to monitor the accumulation of nitrites.

pH
The optimum pH range for Nitrosomonas is between 7.8-8.0.
The optimum pH range for Nitrobacter is between 7.3-7.5
Nitrobacter will grow more slowly at the high pH levels typical of marine aquaria and preferred by
African Rift Lake Cichlids. Initial high nitrite concentrations may exist. At pH levels below 7.0,
Nitrosomonas will grow more slowly and increases in ammonia may become evident. Nitrosomonas
growth is inhibited at a pH of 6.5. All nitrification is inhibited if the pH drops to 6.0 or less. Care must
be taken to monitor ammonia if the pH begins to drop close to 6.5. At this pH almost all of the
ammonia present in the water will be in the mildly toxic, ionized NH3+ state.
Dissolved Oxygen
Maximum nitrification rates will exist if dissolved oxygen (DO) levels exceed 80% saturation.
Nitrification will not occur if DO concentrations drop to 2.0 mg/l (ppm) or less. Nitrobacter is more
strongly affected by low DO than NITROSOMONAS.
Salinity
The nitrifying bacteria in Fritz-Zyme #7 will grow in salinities ranging between 0 to 6 ppt (parts per
thousand) (specific gravity between 1.0000-1.0038).
The nitrifying bacteria in Fritz-Zyme #9 will grow in salinities ranging from 6 up to 44 ppt. (specific
gravity between 1.0038-1.0329).
Adaptation to different salinities may involve a lag time of 1-3 days before exponential growth begins.
Micronutrients
All species of nitrifying bacteria require a number of micronutrients. Most important among
these is the need for phosphorus for ATP (Adenosine Tri-Phosphate) production. The
conversion of ATP provides energy for cellular functions. Phosphorus is normally available to
cells in the form of phosphates (PO4). Nitrobacter, especially, is unable to oxidize nitrite to
nitrate in the absence of phosphates.
Sufficient phosphates are normally present in regular drinking water. During certain periods of the year,
the amount of phosphates may be very low. A phenomenon known as "Phosphate Block" may occur. If
all the above described parameters are within the optimum ranges for the bacteria and nitrite levels
continue to escalate without production of nitrate, then phosphate block may be occurring. In recent
years, with the advent of phosphate-free synthetic sea salt mixes, this problem has become prevalent
among marine aquarists when establishing a new tank.
Fortunately, phosphate block is easy to remedy. A source of phosphate needs to be added to the
aquarium. Phosphoric Acid is recommended as being simplest to use and dose, however, either monosodium phosphate or di-sodium phosphate may be substituted. Fritz PH LOWER contains 31%
phosphoric acid. A one time application of 1 drop per 4 gallons of water is all that is necessary to

activate the Nitrobacter. This small dosage of PH LOWER will not affect the pH or alkalinity of marine
aquaria.
Minimal levels of other essential micronutrients is often not a problem as they are available in our
drinking water supplies. The increasing popularity of high-tech water filters for deionizing, distilling,
and reverse osmosis (hyper-filtration) produce water that is stripped of these nutrients. While these
filters are generally excellent for producing high purity water, this water will also be inhibitory to
nitrifying bacteria. The serious aquarist must replenish the basic salts necessary to the survival of the
aquariums inhabitants. These salts, however, usually lack these critical micronutrients.
Nutriment
All species of Nitrosomonas use ammonia (NH 3) as an energy source during its conversion to
nitrite (NO2). Ammonia is first converted (hydrolyzed) to an amine (NH 2) compound then
oxidized to nitrite. This conversion process allows Nitrosomonas to utilize a few simple amine
compounds such as those formed by the conversion of ammonia by chemical ammonia
removers.
A few strains of Nitrosomonas are also capable of utilizing urea as an energy source. The strains
cultivated for Fritz-Zyme #7 and Fritz-Zyme #9 are among the few that can effectively convert urea.
All species of Nitrobacter use nitrites for their energy source in oxidizing them to nitrate (NO 3).
Color and Smell
The cells of nitrifying bacteria are reddish (Nitrosomonas) to brownish (Nitrobacter) in color. The
solutions in bottles of Fritz-Zyme #7 and #9 are normally peach to rosy colored due to the natural
colors of the bacterial cells and the proprietary solution used to keep them alive. What you see are
actually clumps of bacteria stuck together by their own slime matrix.
Solutions of Fritz-Zyme normally have a musty stagnant water smell.
Sometimes the solution may turn dark brown or black and smell like rotten eggs. This is rare but not
unusual. This is due to the presence of residual sulfates that have been reduced to sulfides. This has no
relationship to the viability of the bacteria. The concentration of sulfides is only a few parts per billion
(ppb) and is not toxic when diluted in the aquarium. These sulfides can be degassed before use if
preferred by removing the bottle cap and allowing oxygen to go into solution. This can be speeded up
by gently aerating the solution for several minutes. The solution will return to its original color. The
dark color and bad odor do not indicate whether or not the product has spoiled.
Light
Nitrifying bacteria are photosensitive, especially to blue and ultraviolet light. After they have colonized
a surface this light poses no problem. During the first 3 or 4 days many of the cells may be suspended
in the water column. Specialized bulbs in reef aquaria that emit UV or near UV light should remain off
during this time. Regular aquarium lighting has no appreciable negative effect.
Chlorine and Chloramines

Before adding bacteria or fish to any aquarium or system, all chlorine must be completely neutralized.
Residual chlorine or chloramines will kill Fritz-Zyme bacteria and fish.
Most US cities now treat their drinking water with chloramines. Chloramines are more stable than
chlorine. It is advisable to test for chlorine with an inexpensive test kit. If you are unsure whether your
water has been treated with chloramine, test for ammonia after neutralizing the chlorine. You can also
call your local water treatment facility.
The type of chloramines formed is dependent on pH. Most of it exists as either
monochloramine (NH2Cl) or dichloramine (NHCl2). They are made by adding ammonia to
chlorinated water. Commercial chlorine reducing chemicals, such as sodium thiosulfate
(Na2S2O2) break the chlorine:ammonia bond. Chlorine (Cl) is reduced to harmless chloride
(Cl- ) ion. Since dichloramine has two chlorine molecules, a double dose of a chlorine
remover, such as sodium thiosulfate, is recommended.
Each molecule of chloramine that is reduced will produce one molecule of ammonia. If the chloramine
concentration is 2 ppm then your aquarium or system will start out with 2 ppm of ammonia. Chlorine
Remover will reduce up to 2 ppm of chlorine at recommended dosages. During the warmer months
chlorine levels may exceed 2 ppm. A double dose would be required to effectively eliminate the excess
chlorine.
Adding Bacteria
After all the chlorine has been safely neutralized Fritz-Zyme should be added to rid the aquarium of
ammonia. Depending on the aquarium pH, 3-4 days may be advisable before adding your fish in order
to minimize stress. If the water supply does not contain chloramines, and there is no ammonia, FritzZyme should be added at the same time as the fish.

Fritz-Zyme #7 (for fresh water) and Fritz-Zyme #9 (for salt water) Biological Water Conditioners are
the sure, safe answer to rapidly establishing the nitrogen cycle within any closed aquatic system. They
are the only concentrated cultures of active autotrophic nitrifying bacteria available and should not be
confused with the many products that are comprised primarily of different species of heterotrophic
bacteria (see our technical bulletin: Nitrifying Bacteria Facts - Autotrophs vs. Heterotrophs).
Fritz-Zyme #7 & Fritz-Zyme #9 contain strains of Nitrosomonas and Nitrobacter species bacteria of the
family NITROBACTERACEAE - the true nitrifiers. Five genera are generally accepted as ammoniaoxidizers and four genera as nitrite-oxidizers. Of these, Nitrosomonas (ammonia-oxidizers) and
Nitrobacter (nitrite-oxidizers) are the most important. Marine species are different from those that
prefer fresh water, and yet, are very closely related. Each species has a limited optimum range for
survival. They are the most efficient, and most important, group of nitrifying bacteria and are
ubiquitous (world-wide) in their distribution.
Fritz Industries pioneered a revolutionary process that decreases their metabolic activity to
allow these bacteria to remain viable for six months in their package. Laboratory tests have

shown that attempts by other competitors to package nitrifying bacteria with extended shelf
lives have failed to produce viable bacteria, and especially, viable Nitrobacter.