Note: Descriptions are shown in the official language in which they were submitted.
36
METHOD AND PRODUCT FOR REMOVAL OF CHLORA~INES,
CHLORINE AND AMMONIA FROM AQUACULTURE WATER
This invention relates to a method and product for water
quality management for aquaculturists. More specifically,
the-invention relates to the simultaneous removal of
chloramines, chlorine and ammonia which are toxic to
aquatic life.
The culture of aquatic organisms, also known as
aquaculture, in the U.S. and elsewhere for food,
recreation, education, research, and hobby purposes is a
fast growing industry. The world production from 1971
to 1978 of fishes, crustaceans and molluscs raised for
food exceeded 10 million pounds. In the U.S. alone these
same species exceeded 184 million pounds in 1980. If
nonfood baitfish and aquarium species were added to the
U.S. production in 1980, then aquacultured animals
would represent more than 206 million pounds. This
translates into a total commercial value of over 210
million dollars. In short, aquaculture is big business
and its growth continues at a significant rate. The
profit potential in aquaculture has created such
incentives that its many enterprises and operations
quickly outdistance the supporting sciences and
technologies This results, not only in production losses
and failure to meet product demands, but also in heavy
financial losses.
Two areas in particular stand out as sources of
frustration and lost profits for aquaculturists. These
are disease management and water quality management.
Although the present invention is particularly related to
water quality management, it should be noted that
chemotherapeutic trea-tment for disease management is
enhanced when high standards of water quality are
maintained.
With fishes and other aquatic animals, just as with
other ani~als and humans, if a proper, non-stressful
environment is provided then the incidence of disease
conditions is all but eliminated. Two major types of
systems exist for raising aquatic life. These are closed
and open systems. There are two types of closed
systems; the closed, recirculating system, and the closed,
non-recirculating system. The closed, recirculating
system, such as a home aquarium, is characterized by a
fixed volume of water which is continuously or
intermittently circulated through a fish holding
tank. The closed, non-recirculating system, such as a
farm pond, is characterized by a fixed volume ~usually
greater than in the previous example) of water to which
fresh makeup water is added, as needed, such as for
compensation for evaporation. In a non-recirculating,
flow-through system, such as a raceway used to raise
trout, fresh makeup water is continuously fed to the fish
holding structure while a like quantity of water is
continuously withdrawn from the structure. The water
environment of the closed, recirculating system can be
controlled with intensive care and maintenance. The water
quality of a closed, non-recirculating system is very
difficult to control except by more or less natural means
(i.e., photosynthetic process to provide oxygen and
bacterial processes to convert toxic wastes). The
non-recirculating, flow-through water systems often have
problems similar to closed systems, but environmental
control in flow-through systems is generally as difficult
--3--
as in closed non-recirculating systems. In either system,
the objective of successful water quality management is
the removal or neutrali~ation of toxic substances which
stress cultured aquatic life forms and thereby to add
significan-tly to the production and profitability of
aquaculture.
Among the many compounds found in natural, waste and
potable waters which are toxic to aquatic organisms,
ammonia (NH3), chlorine in the form of hypochlorus
acid (HOCl) and hypochlorites (OCl ), and chloramines
(NH2Cl, NHC12) are among the most toxic and ubiquitous.
Ammonia is present in natural waters as a result of animal
metabolism of proteins; urinary, fecal and respiratory
wastes; and bacterial mineralization of nitrogenous
bases. This means that the aquatic organisms (fishes,
crustaceans, molluscs, etc.) themselves contribute
significant, toxic pollutants to their own water. In
waste water these same sources, as well as
technological wastes, account for ammonia presence.
Ammonia in potable water is due to the failure to remove
it in the purification process or due to the purposeful
addition for quality control. In a review of management
practices, researcher Stephen Spotte (Fish and
invertebrate culture, John Wiley & Sons, New York, 1979)
observed that current evidence indicates that NH3 is
significantly more toxic than its ionic form, NH4+
(ammonium). Even if this were not true, to attempt to
control the factors such as pH, -temperature and
salinity that effect the NH3:NH4+ ratio could be more
harmful and costly. Spotte suggested that management
techniques be targeted to remove as many sources of
ammonia as possible from the culture water such as uneaten
food, dead animals and plants and to keep the
densities of cultured species moderate and to not allow
the total ammonia level to exceed 0.13 ppm (0.13 mg NH4+
per liter of culture water). The problem with Spotte's
~8~
--4--
suggestions is that for commercial aquaculturists moderate
densities of cultured animals are seldom profitable and
for aquarium hobbyists there is always room for one more
fish in an already overcrowded aquarium.
other respected researchers have warned of the dangers of
ammonia in aquaculture. The safe level for salmonids such
as trout and salmon is considered to be from 0.005 to 0.02
ppm. As a predisposing factor in bacterial gill disease
of cultured food fishes, ammonia levels over 0 3 ppm
are considered dangerous.
~hen the foregoing standards for water quality are
compared to ammonia levels which can be encountered in
both culture water and natural waters, the serious
nature of this problem can be better appreciated. The
ammonia levels in wastewater can range up to and over 5000
ppm. In aquarium and aquaculture systems it is not
unusual to encounter total ammonia concentrations of
between 1.0 and 3.0 ppm.
Another fish threatening substance is chlorine. Chlorine
is most often present in water as a result of disinfection
processes. It is not found in natural waters unless there
has been contamination from wastewater or potable
water sources. Aquaculturists and aquarists simply have
no direct control over the quantity of chlorine, or
associated chloramines, introduced to municipal water
supplies. However, no matter what the initial
concentration of chlorine, or chloramines, it must be
reduced to zero before any water in which it is present
can be safely used for culture purposes. Levels of
chlorine Erom 0.2 to 0.3 ppm are rapidly toxic to
fishes. The U.S. Environmen-tal Protection Agency
recommends an upper level of 0.003 ppm for continuous
exposure by coldwater and warmwater fishes. Chlorine
levels in municipal water supplies range up to 2.5 ppm.
When used as a disinfectant agent for cleaning aquariums,
the recommended solution typically contains 50 ppm
chlorine. Accordingly, the aquarium must be very
thoroughly rinsed to remove any trace of the chlorine
after cleaning.
Chloramines are most often present in wa-ter for the same
reasons as the presence of chlorine. However, some
chloramines in natural and waste waters result from the
chemical combination of chlorine with the ammonia normally
found in these waters. The chloramine level in a
given water can range quite high to over 5000 ppm, but the
levels encountered in most municipal tap waters is in the
range of 0.5 to 4.0 ppm. Even the latter range represents
a deadly concentration level for aquatic life.
The reduction in concentrations of these toxic components
in water, when their initial introduction cannot be
controlled, is crucial in the culture, maintenance and
display of freshwater, brackish water (estuarine), and
marine organisms. In addition, the timely reduction
in concentrations of these toxic components is also
desirable.
In the case of chlorine a process of reductive
dechlorination is most often practiced. However,
granular activated carbon is also used as a chemical
adsorbant to remove chlorine from water. Ammonia removal
can be accomplished by adsorption on zeolites like
clinoptilolite and phillipsite and by bacterial
nitrification. The efficiencies of these two
processes are effected by contact time (i.e., how long the
water is in contact with -the adsorbant or bacterial bed)
and other conditions such as temperature, dissolved oxygen
levels, the presence of interferring substances (i.e.,
certain antibiotics in -the case of nitrifying
bacterial beds, and highly surface active organics in the
case of chemical adsorbants), and maintenance procedures
(i.e., cleaning routines) of the filters themselves.
86
.
--6--
Chloramines can be removed by reductive dechlorination
followed by adsorption or nitrification of the freed
ammonia.
Dechlorination has been shown to be a highly reliable
process and one which works well under most conditions
found in culture water, the term referring to the water
used to maintain, grow or breed aquatic plants and
animals. One problem with this process presents itselE
when thiosulfates, S2O3 (the substances used in the
majority of commercially available dechlorinators), are
used; excess thiosulfate ion reacts with dissolved oxygen
in water and inadvertent or purposeful overdosing can
result in a reduction of dissolved oxygen in culture water
which can in turn cause respiratory stress in the
cultured organisms. In addition, many commercially
available dechlorinators have been found to be inadequate
for complete dechlorination of even relatively lightly
chlorinated (i.eO, less than 4.0 ppm total chlorine)
potable waters. The use of granular activated carbon
has been common and is most often employed in laboratories
for the preparation of chlorine-free culture water.
Nevertheless, a recent study details the problems
associated with using granular activated carbon alone as a
method of dechlorinating water for aquatic
toxicological studies. Stephen J. Mitchell and Joseph J.
Cech, Jr., 1983, "Ammonia-caused gill damage in channel
catfish (Ictalurus punctatus): confounding effects of
residual chlorine", Can. J., Fish. Aquat. Sci., 40(2),
pp. 242-247.
The elimination of chloramines (i.e., dechloramination)
from water used for culture purposes has reached more
creative levels. One method currently used is to
d~chlorinate with the usual dechlorinators then remove
the freed ammonia by adsorption on granular clinoptilolite
placed in a filtering device or by addition the finely
divided, powder, clinoptilolite directly to the water. In
~:~2~6;
--7--
actuality, most dechloramination is achieved by
dechlorinating in the usual way and allowing the ammonia
to be oxidized by nitrifying bacteria. Just as with
chlorine, granular activated carbon has been used to
remove chloramines from water, but this process has
also been questioned by the Mitchell and Cech study in
1983. The same study showed that partial dechlorination
allowed the residual chlorine to potentiate the toxic
effects of ammonia on fish.
The removal of ammonia released into water when
chloramines are dechlorinated is likewise problematical.
Biological filtration is a process of bacterial conversion
or nitrification of toxic ammonia and nitrite ions (NO2 )
to less toxic nitrate ions (NO3 ). siological
filtration, however, is easily interrupted and inhibited,
and the intermediate product, nitrite ions (NO2-), is
significantly more toxic to aquatic organisms than the
precursor ammonia. Until a biological filter bed is fully
conditioned and properly functioning (an average of 21
days), there is a constant increase in the concentration
of the nitrite ions until the precursor ammonia is reduced
below its inhibiting (to the nitrite converting
Nitrobacter species of bacteria) concentrations.
The removal of ammonia by adsorption has its own set of
problems. Among these are flow rate and contact time,
adsorbant grain size, temperature, adsorbant capacity) and
the concentration of interfering ions such as sodium (Na~)
and potassium (K+). Clinoptilolite has approximately
5% of the capacity in salt water that it exhibits in
freshwater and, therefore, correspondingly larger
quantities of this adsorbant are required for a salt water
application.
Commercial products do currently exist which claim to
remove or neutralize one or more of the toxic substances
chlorine, chloramines and ammonia. However, they all
~.~1~2~
suffer from one or more of the above described
shortcomings. Simple dechlorinators, if properly dosed
will completely neutralize chlorine. These same
dechlorinators will break the chlorine-ammonia bonds in
chloramines and neutralize the chlorine but will not
neutralize the freed ammonia. Ammonia adsorbants, if used
properly will adsorb and remove ammonia from water, but
will have no effect upon any chlorine present and they
will not function properly in saline waters. Biological
filters will maintain ammonia concentrations at low
levels, even in saline waters, but they require a long
start-up time (up to 21 days), are slow to react to
increased ammonia loads, and require a relatively narrow
range of operating conditions. Products which are
combinations of dechlorinators and ammonia adsorbants
will not function in saline waters and cause a temporary
cloudiness in the treated water due to the dispersion of
the finely divided adsorbant.
Accordingly, aquaculture needs a safe and effective
way to remove chloramines, chlorine and ammonia which
overcomes the limitations, dangers and shortcomings of the
various techniques presently employed. The primary goal
of this invention is to fulfill this need in the industry.
More specifically, an object of the invention is to
provide a product and method for the removal of
chloramines, chlorine and ammonia which, unlike existing
zeolites and ion-exchange resins, functions as well in
saline water as it does in freshwater treatment.
Ano-ther object of the invention is to provide a completely
safe product and method for the removal of chlorine,
chloramine and ammonia which is non-toxic to fishes~
aquatic invertebrates, marine and freshwater algaes,
and aquatic plants.
Another object of the invention is to provide a product
~1~2~;
g
and method for the removal of chloramines, chlorine and
ammonia which does no-t cause clouding in hard or soft
water or in salt water as do products which contain
insoluble zeolites.
S
Yet another object of the invention is to provide a
product and method for the removal of chloramines,
chlorine and ammonia in which the time required for
neutralization is greatly reduced from the time required
by earlier techniques. With this invention,
neutralization times vary from one to five minutes for
"free" chlorine (hypochlorites), ten to thirty minutes for
chloramines (ncombined" chlorine), and twelve minutes to
one hour for free ammonia.
An additional object of the invention is to provide a
product and method for the removal of chloramines,
chlorine and ammonia which does not react with dissolved
oxygen in either freshwaters or saline waters.
Another object of the invention is to provide a product
and method for the removal of chloramines, chlorine and
ammonia which is not pH dependent and functions equally
well throughout the "normal" pH range, 5.0 to 9.0, of
waters in which most aquatic life is found.
Another object of the invention is to provide a product
and method for the removal of chloramines, chlorine and
ammonia which is largely uninhibited by the presence of
commonly used antibiotics such as chloramphenicol,
nitrofurans, and sulfa drugs, or by the presence of
antiparsiticals such as copper sulfate, metronidazole and
formaldehyde.
A further object of the invention is to provide a
product and method for the removal of chloramines,
chlorine and ammonia which can be combined with water
conditioning chemicals such as other dechlorinators,
--10--
electrolyte mixes, and trace element mixes.
Yet a further object of the invention is to provide a
product and method of the character described which is
safe, reliable and economical to effect removal of
chloramines, chlorine and ammonia.
Other and further objects of the invention, together with
the features of novelty appurtenant thereto, will appear
in the course of the following description of -the
invention.
I have discovered that an alkali metal
formaldehydebisulfite effectively neutralizes chloramines,
chlorine and ammonia from saline and fresh waters for
use in aquaculture. A pure alkali metal
formaldehydebisulfite, a mixture of alkali metal
formaldehydebisulfites, or a mixture of one or more alkali
metal formaldehydebisulfites with various diluents,
carriers or other ingredients can be utilized directly
in untreated water to neutralize by chemical reaction any
aqueous chloramines, chlorine and ammonia which may be
present in order to render the water nontoxic for aquatic
life. The water treatment product can be manufactured
either in a dry form (i.e., powder, granule, flake,
tablet, cake, pellet, bolus, capsule) with or without
additives, or in a water solu-tion with or without other
dissolved or suspended substances.
In summary, the invention relates to a process for
neutralizing chloramines, chlorine and ammonia in water by
adding an alkali metal formaldehydebisulfite in a dry or
solution form in which the alkali metal
formaldehydebisulEite is selected from the group
consisting of sodium formaldehydebisulfite and
potassium formaldehydebisulfite. In a preferred
embodiment, the alkali metal formaldehydebisulfite is
sodium formaldehydebisulfite added in the amount at least
equal to the greater of the quantity required to react on
a one to one molecular basis with a reactant selected from
the group consisting of 4 times the stoichiometric amount
oE ammonia, 12 times the stoichiometric amount of
monochloramine, 10 times the stoichiometric amount of
dichloramine and 12 times the stoichiometric amount of
chlorine in the form of hypochlorites present in the water
to be treated.
Unless otherwise stated herein, indication of parts or
percentages are given on a weight basis.
I have discovered that an alkali metal
formaldehydebisulfite is unexpectedly effective and safe
to neutralize chlorine, chloramine and ammonia from
saline and fresh waters for use in aquaculture. A pure
alkali metal formaldehydebisulfite, a mixture of alkali
metal formaldehydebisulfites, or a mixture of one or more
alkali metal formaldehydebisulfites with various diluents,
carriers or other inert ingredients can be utilized
directly in dry or solution form in untreated water to
neutralize by chemical reaction any aqueous chloramines,
chlorine and ammonia which may be present in order to
render the water nontoxic for aquatic life.
Neither the reaction mechanism, nor the reaction products,
by which the phenomena of neutralizing chloramines,
chlorine and ammonia in water with an alkali metal
formaldehydebisulfite is understood. However,
experimental research shows reaction completion of
sodium formaldehydebisulfite and the representative target
compounds including free chlorine in the form of sodium
hypochlorite (NaOCl), free ammonia (NH3), and
monochloramine (NH2Cl) to eliminate the toxic effects of
the target compounds in aquaculture. The results show
that under conditions of varying pH, hardness, and
salinity that sodium formaldehydebisulfite was capable of
simultaneously reducing the concentrations of all three
286
- 12 - 61316-657
representative target compounds to safe levels. Further research
indicates effectiveness under representative aquaculture working
conditions by reducing free ammonia levels ln existing freshwater
and marine aquarium water and by neutralizing free chlorine and
chloramines in freshly drawn potable tap water.
This invention is suitable for use in any natural or
cultured water system supporting aquatic organisms. This process
is especially suitable for neutralizing pollutants in aquaculture
water having a pH of about 6 - 9.
As used herein, the terms "remove" and "neutralize" are
used interchangeably to refer to the discovered ability of alkali
metal formaldehydebisulfites to render nontoxic to aquatic life
the chloramines, chlorine and ammonia existing in natural and
culture waters.
The alkali metal formaldehydebisulfites useful in this
invention include sodium formaldehydebisulfite and potassium form-
aldehydebisulfite. The compound sodium formaldehydebisulfite has
the chemical formula HOCH2S03Na, and is also known as formaldehyde
sodium bisulfite and sodium hydroxymethane sulfonate. The
compound potassium formaldehydebisulfite has the chemical formula
HOCH2SO3K, and is also known as formaldehyde potassium bisulfite
and potassium hydroxymethane sulfonate.
The alkali metal formaldehydebisulfites may be utilized
in the dry form with a variety of inert materials such as
diluents, carriers, axcipients, lubricants, disinter~rants, and
colorants. A diluent (i.e. tricalcium phosphate) is an inert
material used to reduce the concentration of an active material to
achieve a desirable and beneficial effect. A carrier (i.e., salt)
~130~8~;
- 12a - 61316-657
is an inert material used to deliver or disperse an active
material.
Suitable diluents and carriers for use with alkali metal
formaldehydebisulfites include salt and other similar, non-
reactive, neutral electrolytes such as sodium ~ulfate and
potassium chloride, and non-electrolytes and insoluble salts such
as starch, sugars, clays, and calcium
'E3~
- ~a30~KE8~;
-13-
sulfate. An excipient (i.e., starch) is an inert material
used as a binder in tablets. Suitable excipients for use
with alkali metal formaldehydebisulfites include polymers
and gums such as cellulose gum and povidone, and
starches. A lubricant (i.e., magnesium stearate) is
an inert material used to reduce friction during filling
or tableting processes. Suitable lubricants for use with
alkali metal formaldehydebisulfites include fatty acid
salts such as calcium stearate or magnesium stearate, and
paraffinic compounds and fatty acids such as paraffin
wax and stearic acid. A disintergrant is an inert
material that causes tablets and boluses to burst upon
exposure to appropriate conditions. Suitable
disintergrants for use with alkali metal
formaldehydebisulfites include polymers such as cross-
linked povidone, and effervescent mixtures such as sodium
bicarbonate/citric acid. A colorant is an inert material
which imparts color to another material or mixture.
Suitable colorants for use with alkali metal
formaldehydebisulfites include lakes (i.e., organic
pigments on an adsorptive inorganic substrate) such as
` rose madder, and non-oxidizing dyes such as acriflavine.
The pure, dry active alkali metal formaldehydebisulfite
may be packaged in containers such as bottles, boxes,
non-porous bags, and drums which serve to protect the
integrity of the product and to allow for appropriate
dispensing. Other dry forms of the product for use in
this invention include unit dose tablets, capsules,
boluses or packets. Individual dosage units may be
weighed or measured by bulk volume if supplied in the form
of powders, granules, pellets, or flakes. Larger uni-t
dose requirements, such as for ponds, lakes, or streams
can be applied as a dry-manufactured cake.
The alkali metal formaldehydebisulfite may be used
directly for water trea-tment. As with many process
chemicals, however, proper perfo-rmance of a consumer
~3~W2~1~
-14-
formulation is often best achieved by providing a dosage
form which is easily applied and only requires commonly
available volumetric measuring devices such as teaspoons
(l/6 fl.oz, 4.93 mL), tablespoons (l/3 fl.oz., 9.86 mL) or
cups (8 fl.oz., 236.64 mL). Although weight
measurement of solids is routinely very accurate, it is
quite uncommon for consumers to employ such measurements
when using water conditioning chemicals and similar
products. The use of pre-weighed unit dosages of a pure
substance or its mixtures in the form of tablets,
boluses, capsules or packets is the typical and preferred
method to deliver dry forms. Such method works quite well
for this invention. Equally satisfactory, however, is a
formulation as a dry powder or granular mixture of a
consistency which readily lends itself to accurate and
repeatable volumetric dosage measurements.
To illustrate the foregoing principals with respect to
suitable formulations, the following dry forms of product
represent convenient formulations adapted for easy use
by the lay consumer:
(l) Unit-dose tablet containing 1.18 grams of active
ingredient sodi-um formaldehydebisulfite and 0.80 grams of
diluent salt and 0.02 grams of lubricant magnesium
stearate designed to treat 10 gallons (37.8 liters) of
municipal water containing up to 2.5 ppm (2.5 mg/liter) of
chlorine as monochloramine.
(2) Multi-dose package containing l pound (453.6
grams) of a mixture of 9.44 ounces (267.6 grams) of sodium
formaldehydebisulfite and 6.56 ounces (186.0 grams) of
fine blending salt which is to be dosed at the rate of 1
teaspoonful (approximately 5 mL or 10 grams) per lO
gallons (37.8 liters) of aquarium water, either freshwater
or marine, containing up to l.0 ppm (l.0 mg/liter) of free
ammonia (NE~3).
The formulation of the aquaculture product in solution
form, whether by direct dissolution of the active
~31~2~;
--15--
ingredient or in situ synthesis of the product by reaction
of formaldehyde gas or solution with sodium bisulfite
solution at the the point of manufacture or just prior to
being added the water to be treated is inherently simple,
5 convenient and inexpensive. Highly purified sodium
formaldehydebisulfite is commercially available in large
quantities. It is easily and completely soluble in water
and produces a solution which is clear, colorless,
odorless and stable within a pH range of 6.0 to 8Ø
To illustrate the foregoing principals with respect to
suitable liquid formulations, the following solution forms
of product represent convenient formulations adapted for
easy use by the lay consumer:
(1) A two-part product consis-ting of a 9.525%
solution of formaldehyde (CH2O) in water with or without
suitable preservatives and/or buffers (i.e., methanol,
phosphate buffer) and a second solution of a 33.01~6
solution of sodium bisulfite (NaHSO3) in water. The two
20 solutions to be combined, in equal portions by weight,
and used at the rate of 5 mL per 10 gallons (37.8 liters)
to treat pond water containing 1.0 ppm (1 mg/liter) free
ammonia.
(2) A single solution containing 21.279~ sodium
25 formaldehydebisulfite in water, to be used at the rate
of 1 teaspoon (approximately 5 mL) per 20 gallons (75.7
liters) for the treatment of potable water containing 2.0
ppm (2.0 mg/liter) combined chlorine.
30 The consumer oriented single solution may be used as a
dose of 1 teaspoonful (4.93 mL) per 20 gallons (75.71) of
water for neutralizing up to 4.0 ppm monochloramine
measured as combined chlorine, or 1 teaspoonful per 10
gallons (37.85 L) for neutralizing up to 1 ppm ammonia
35 measured as free ammonia. In each instance, the
recommendecl dilution represents approximately 4 times the
required stoichiometric amounts oE sodium
formaldehyclebisulfite required to reac-t with the
13~0286
- 16 - 61316-657
pollutants. Such a product is perfectly capable of neukralizing
chlorine in the form of hypochlorites without the addition of any
other dechlorinator. Such a product would not be expected to
neutralize any ammonia present as the ammonium ion. However, this
of course is relatively unimportant because of the non-toxic
nature of the ionized form. Reaction with chlorine in both the
"free" and "comhined" forms can be expected to proceed as quickly
as with all-purpose water conditioners (i.e., complete within 10
minutes). The reaction with ammonia can be expected to take
longer depending upon the initial concentration of the free
ammonia. In usual practice, complete deamination can be expected
within 1 hour a~ an initial concentration of 1 ppm ammonia.
Unlike products containing mixtures of thiosulfates and
clinoptilolite, sodium formaldehydebisulfite solutions function in
saltwater as easily as in freshwater, and there will be no
clouding. The sodium formaldehydebisulfite solution is of
necessity colorless because of its incompatibility with aryl-
methane and other similar oxidizing dyes. Methylene blue, a
sulfur-containing, oxidizing dye is stable in a sodium
formaldehydebisulfite solution for a short period of time, but
because of the objectionable properties of this dye, it is not
recommended as a coloring agent. It is most preferable that the
aquaculture water in which the sodium formaldehydebisulfite is
added be free of oxidizing pollutants aside from those pollutants
that are targeted in this invention. Preferably the aquaculture
water will be free of oxidizing pollutants selected from the group
consisting of permanganates, peroxides dichromates arylmethane
dyes, and mixtures thereof.
~L30~286
- 16a - 61316-657
In the various product formulations of this invention,
the usual practices of cleanliness and sanitation are to be
follo~ed as these relate to the conditions of mixing, packaging
and storage. The purity and grades of the various ingredients
including formaldehyde gas, formalalehyde solution, sodium
bisulfite, sodium formaldehydebisul.fite, potassium
formaldehydebisulfite, methanol, salt, starch, tableting
excipients, tableting lubricants, cross-povidone, buffers and
their components, colorants and dyes, diluents and carriers, and
water may vary from standard commercial or technical
, ~.~.,
.~
~:~0(:~286
-17-
grades to the highly purified, reagent or pharmaceutical
grades. The physical forms of the various solid
components may vary from ultra-fine powders to granular
and flake forms. Liquid components should be free from
suspended or precipitated material, but should such
material be present it shou:Ld be removed by suitable
settling and decantation or filtering. Any water should
be similarly free of suspended or precipitated material as
well as free from free or combined chlorine, including
chloramines, and free or ionized ammonia. In
addition, the water must be free of other free halogens
such as iodine (I) and bromine (Br) and their combined
forms which may reduce the final required concentration of
the sodium formaldehydebisulfite. Neutralization or
removal of potentially interfering substances would be
- permissible prior to the addition or formation of the
sodium formaldehydebisulfite. With observance of the
foregoing, any potable water source may be used in the
manufacturing process.
All components used in the production of these aquacul-ture
products must be free from, or rendered free from,
substances which may be toxic or otherwise detrimental to
fish, aquatic invertebrates, aquatic algae plants and
other aquatic life.
Both acids and alkalies hasten the decomposition of the
sodium formaldehydebisulfite. Therefore, solutions of
sodium formaldehydebisulfite must be manufactured so that
the final pH lies between 6.0 and 8Ø Dry
formulations must be manufactured so that if the resul-tant
mixture is hygroscopic, then decomposition of the sodium
formaldehydebisulfite will not result due to an acidic or
alkaline environment being created within the mixture.
In typical production of a liquid form of this invention,
the following parameters should be considered preferable
but not essential. If dry sodium formaldehydebisulfite is
~30~Z86
-18-
to be used it should be of photographic grade which has
typically less than 0.1~ free formaldehyde and less than
0.1% uncombined sodium bisulfite and of powdex form. If
the sodium formaldehydebisulfite is to be formed by
reaction, then 35-38~ formaldehyde solution with no
more than 15% methanol can be reacted with photographic
grade, powdered or granular sodium bisulfite. Water
should be completely deionized and have a pH of 6.5 to
7.5. The final solution is to be clear, colorless and
free from suspended or precipitated matter. Mixing
and filling equipment may desirably be fabricated from
stainless steel or polyvinyl chloride. The liquid product
can be packaged and stored in polyethylene containers with
polyethylene or polypropylene closures.
The relative instability of formaldehydebisul~ites in
alkaline media does not recommend the use of the in situ
reaction of formaldehyde solution with alkali sulfites,
and the presence of amounts of hydroxide ions (OH )
equimolar to the amounts of alkali metal
formaldehydebisulfites could be detrimental to aquatic
life in treated waters due to an increase in pH which
might result.
As previously indicated, the reaction mechanism and
reaction products are not completely understood in the
reaction of alkali metal formaldehydebisulfite with
chloramines, chlorine and ammonia in water. Howevert
research indicates that the reactants react on a one to
one molecular basis and this observation is utilized
in formulating the amount of alkali metal
formaldehydebisulfite in a dry or solution form which is
required for a specified amount of pollutant found in the
water to be treated. Accordingly, as minimum effective
quantities, alkali metal formaldehydebisulfite must be
present in an amount at least equal to the greater of the
quantity required to react on a one to one molecular basis
with the stoichioinetric amount of ammonia, the
~3[)~28~ 1
--19--
stoichiometric amount of monochloramine, the
stoichiometric amount of dichloramine or the
stoichiometric amount of chlorine in the form of
hypochlorites present in the water to be treated. It is
S naturally desirable, however, that the neutralizing
agent be present in excess. In a preferred embodiment of
the invention, the alkali metal formaldehydebisulfite is
sodium formaldehydebisulfite added in the amount at least
equal to the greater of the quantity required to react on
a one to one molecular basis with a reactant selected
from the group consisting of 4 times the stoichiometric
amount of ammonia, 12 times the stoichiometric amount of
monochloramine, 10 times the stoichiometric amount of
dichloramine and 12 times the stoichiometric amount of
chlorine in the form of hypochlorites present in the
water to be treated.
The invention is further exemplified with reference to the
following research examples investigating the parameters
of the alkali metal formaldehydebisulfites for use in
aquaculture and the efficacy for neutralizing chloramines
chlorine and ammonia from water to provide a safe
treatment program for aquatic life in culture water.
Example 1
Research was conducted to investigate the reaction between
sodium formaldehydebisulfite (hereinafter referred to as
"SFB" ) and ammonia as represented by the following
equation:
SFB + NH3 ----> ????
To test this reaction I prepared ammonia standards from
ammonium chloride and used the standard method of
measuring for ammonia with an Orion 901 Ionalyzer
connected to a chart recorder and an ammonia specific-ion-
electrode (hereinafter referred to as "SIE"). Two working
solutions of SFB, #1 SFB solution (120.85 g/L) and #2 SFB
~,3~2~16
-20-
solution (241.72 g/L) were prepared for use in the
subsequent reaction studies. The s-tandard ammonia
solutions were made so that upon addition of 1 mL of 10N
sodium hydroxide (NaOH) concentrations of 0.5 ppm, 1.0 ppm
and 2.0 ppm (as NH3) were produced. One-hundred
milliliters of each standarcl ammonia solution were
pipetted, in turn, into 150--mL Fleakers, a Teflon-coated
stirring bar was placed in the Fleaker and gentle stirring
was started. The prepared and standardized SIE was placed
into the solution and when a stable baseline was
achieved on the recorder, 1.0 mL of the NaOH solution was
pipetted into the solution. As soon as a stable reading
was obtained on the instrument and a new, stable baseline
was achieved on the recorder then an excess (1.0 mL) of
the #1 SFB solution was added and any change in the
measured ammonia concentration was tracked on both the
instrument and the recorder with instrument readings being
noted on the recorder chart at arbitrar~ intervals.
The results of the chemical tests suggest that the
reaction between ammonia and SFB even at high pH's is a
concentration dependent, second order reaction. As such
the reaction time is limited by the concentrations of both
reactants. This means that very low concentrations of
ammonia (less than 0.5 ppm) will exhibit very long
reaction times. In my tests a 10% reduction in the
initial concentration (0.5 to 0.45 ppm) took 15 minutes as
illustrated in Table 1. A similar percentage reduction
took only 3.68 minutes at an ini-tial concentration of 1.0
ppm with a change in rate at 15.26 minutes
(concentration of approxima-tely 0.65 ppm) as shown in
Table 2. With reference to Table 3, and with a 2.0 ppm
initial ammonia concentration, a 10~ reduction was
achieved at 4.88 minutes with a rate change at 14.1
minutes (concentration of approximately 1.40 ppm) and
an approximate 50~ reduction (1.06 ppm) was achieved at
33.16 minutes. In each case, subsequent additions of the
#l SFB solution resulted in an increase in the reaction
~a3~28~
-21-
rate.
Table 1.
Time: Conc. (ppm):
0:00.00 0.500
0:05.08 0.491
0:07.06 0.484
0O09.04 0~476
0:11.06 0.470
0:13.00 0.461
0:15.00 0.450
Table 2.
Time: Conc. (ppm):
0:00.00 1.00
0:02.00 0.938
0:03.68 0.900
0:05.96 0.833
0:06.96 0.810
0:08.00 0.788
0:10.50 0.729
0:14.40 0.656
0:23.10 0.590
0:32.80 0.531
-- i90~;28~;;
-22-
Table 3.
Time: Conc. (ppm):
0:00.00 2.00
0:02.00 1.97
0:04.88 1.80
0:07.40 1.62
0:11.12 1.46
0:17.00 1.31
0:23.70 1.18
0:33.16 1.06
0:33.52 Second Addition of SFB
0:34.98 1.00
0:36.20 0.955
0:39.42 0.861
0:41.80 0.800
0:43.28 0.770
0:44.00 Third Addition of SFB
0:45.06 0.697
0:46.76 0.627
0:48.98 0.565
0:51.78 0.508
0:56.18 0.457
0:56.98 0.450
Example 2
Additional tests were conducted to confirm the react~n
between SFB solutions and ammonia. In the first test, an
ad libidum solution of ammoniacal silver nitrate was
prepared by dissolving an unweighed quan-tity of silver
nitrate (AgNO3) in just enough 6.0N ammonium hydroxide to
produce a clear colorless solution free of precipitate.
Since it is known that any reduction in the slight
excess oE ammonia in this solution will result in the
precipitation of an insoluble silver compound, it was
assumed that just such a precipitate would form if SFB or
~0~
-23-
its solutions were added to the solution. Side-by-side
controls were used to con-firm that any reaction was not
due to some other variable such as loss of ammonia from
the solution to the atmosphere. Identical test tubes were
used. In each of six tubes, 15 drops of the
ammoniacal silver nitrate solution were added. Then 10
drops of totally deionized water were added to each tube
and each tube, in turn, was swirled to mix. In three of
the tubes, 25 drops of #2 SFB solution was added to
each. In the remaining tubes, 25 drops of totally
deionized water was added to each. Each tube was swirled
to insure mixing of the contents, and allowed to stand to
observe any visible changes.
In the test series of the ammoniacal silver nitrate
with the SFB solution the three tubes without added SFB
solution showed no reaction during the test which was
terminated after 1 hour. The three tubes with added SFB
solution showed an immediate reaction (within 30 seconds);
a white precipitate was formed which coagulated
readily upon additional swirling of the tubes. In
addition, the SFB-containing tubes exhibited only a slight
musty odor while the untreated tubes exhibited a
noticeably stronger and distinct ammonia odor. This test
showed that the SFB reacted to remove the ammonia
allowing the formation of an insoluble silver precipitate
compound.
Example 3
In a test series further examining the ammonia and SFB
reaction, 25 drops of 1.0 M cupric sulfate solution was
added to each of six identical test tubes. Next, 25 drops
totally deionized water was added to each tube and each
tube was swirled to insure mixing of the contents.
Next, 15 drops of 6.0N ammonium hydroxide solution was
added to each tube to produce a solution of ammoniacal
cupric sulfateO As with the ammoniacal silver ni-trate,
86
-24-
any dissipation or removal of ammonia from this solution
will result in the precipitation of an insoluble copper
compound. Finally, 50 drops of #2 SFB solution was added
to three of the six tubes, and 50 drops of totally
deionized water were added to the remaining three
tubes. Each tube was swirled to mix the contents.
The series of tests with ammoniacal cupric sulfate showed
essentially the same results as the test with ammoniacal
silver nitrate. The color change of the solutions and
the color of the precipitate were more dramatic. In the
SFB-containing tubes, the clear, bright blue solution
became turbid within 5 to 7 minutes and within 2 hours the
color of the solution changed to a very pale blue and a
copious amount (equivalent to approximately 1/3 the
volume of the total liquid in the tube) of blue-green
precipitate had formed. The control tubes exhibited no
color changes or precipitate formation during the test
which terminated after 4 hours. As with the previous test
series, only a slight musty odor was detectable in the
SFB-containing tubes while in the control tubes a strong
and distinct ammonia odor was detectable.
The same test parameters were employed in which two
different solutions were substituted for the #2 SFB
solution. These two solutions were: 1) an SFB solution
(hereinafter "#3 SFB solution") in which equal volumes of
sodium bisulfite having a 1 M concen-tration as sulfur
dioxide and a 1 M formaldehyde solution were mixed; and 2)
a potassium formaldehydebisulfite (hereinafter "KFB")
solution in which equal volumes of potassium metabisulfite
(K2S2O5) having a 1 M concentration as sulfur dioxide and
a 1 M formaldehyde solution were mixed. Both solutions
were designed to be 0.5 M in the respective alkali metal
formaldehydebisulfite (i.e., sodium in the former and
potassium in the la-tter solution). When tested in a
manner equivalent to that of the #2 SFB solution above
using 90 drops of each solution, identical results were
~3~28~;
-25-
obtained with solutions of ammoniacal cupric sulfate.
In an additional test series, a combination solution of
equal volumes of #3 SFB solution and KFB solution were
substituted for the #2 SFB solution at the rate of 45
drops of each of the ~3 SFB and KFB solutions, added
together, to solutions of ammoniacal cupric sulfate. This
combination solution reactecl in an identical manner as
cited above for the #2 SFB solution.
Example 4
In the next test series, 15 drops of 6.0 N ammonium
hydroxide was added to each of six test tubes. Next, 3
drops of bromthymol blue, U.S.P. test solution used as
a pH indicator, was added to each tube, and each tube was
swirled to mix the contents. Each solution turned a
characteristic blue color indicating an alkaline pH.
Next, 50 drops of #2 SFB solution was added to three of
the tubes and 50 drops of totally deionized water the
the remaining three tubes. Each tube was swirled to
insure mixing of the contents and each tube was stoppered
with tightly fitting silicone rubber stoppers and allowed
to stand to observe any changes in the color of the
solutions. One tube containing the added SFB solution
and one tube with just added water was periodically opened
at intervals of every 15 minutes for the first 2 hours,
then every hour for 6 additional hours, and smelled to
test for the odor of ammonia. The other four tubes
remained unopened throughout the entire 8 hours of the
test.
Throughout the test, all tubes retained the color
indicating an alkaline pH. In the control tubes, the
stoppers were easily removed indicating no reduction
in the atmospheric pressure within the tubes. In the SFB-
containing tubes, the stoppers in the two tubes which
remained unopened during the test had been pulled
~30~28~
-26-
noticeably deeper into the mouth of the tube to a depth of
.5 to .7 centimeters greater than the stoppers in the
other four tubes. Greater effort was required to extract
the stoppers from these two tubes. In the one SFB-
containing tube which was periodically opened for odortesting there was no noticeable difference in the depth of
the stopper or in the effort required to extract it at the
termination of the test. There was a clearly detectable
difference in ammonia odor in the three SFB-containing
tubes compared to the odor in the control tubes at the
termination of the test. The reduction of the atmospheric
pressure in the control tubes is indicative of the
consumption of ammonia, which ordinarily has an
appreciable vapor pressure at room temperature. The
initial odor of each tube was the characteristic
pungent odor typical of ammonia solutions.
Example 5
Research was conducted to investigate the reaction
between sodium formaldehydebisulfite (hereinafter referred
to as "SFs") and monochloramine as represented by the
following equation:
SFB + H2NCl ----> ????
The experiment was designed so that any reduction in
either chlorine or ammonia concentrations or both could be
observed, rather -than speculate as to the nature of the
reaction products. I prepared a standard monochloramine
solution from 28% ammonia solution and sodium hypochlorite
solution so that the resulting solution contained l
ppm monochloramine (H2NCl)~ One-hundred milliliters of
this solution were pipetted into a 150-mL Fleaker and
prepared as for the ammonia tests given in prior Example l
except no NaOH solution was added. When an instrument
reading of 1.00 ppm was achieved, 1.0 mL of the #l SFs
solution was added and any change was tracked on both the
instrument and the recorder with instrument readings being
noted on the recorder chart at arbitrary intervals.
~3002 !36
-27-
In the chloramine reaction test, the readings as shown in
Table 4 indicated an increase from the "set" meter reading
of lo 00 ppm to 1.46 ppm at 3.98 minutes at which time the
measured concentration of ammonia started decreasing.
This is consistent with a first order dechlorination
reaction followed by reaction with ammonia. As in
previous Example 1, addition of more of the SFB solution
resulted in an increased reaction rate. From the first
point oE recorded decrease in the ammonia concentration to
the point of the second addition of SFB solution, the
rate was equivalent to approximately 0.91% decrease in
; ammonia per minute. From the second addition to the third
addition of SFB solution, the rate was approximately 4.7~
decrease per minute, and from the third addi-tion, the rate
was approximately 7.4% decrease per minute.
~3~2~q~
-28-
Table 4.
Time: Conc. (ppm):
0: 00. 00 1. 00
0:03.98 1.46
0:05.55 1.44
0:07.19 1.42
0:08.53 1.40
0:10.00 1.38
0:11.75 Second Addition of SFB
0:12.15 1.25
0:13.30 1.20
0:13.98 1.15
0:14.72 1.10
0:15.45 1.05
0:16.~5 1.00
0:17.55 0.950
0:18.15 0.900
0:19.80 0.850
0:22.85 0.750
0:27.09 0.650 Third Addition of SFB
0:28.75 0.600
0:29.55 0.500
0:31.8~ 0.400
0:35.51 0.300
0:36.80 0.275
Example 6
Research was conducted to investigate the reaction of SFB
solutions with tap water, conditioned (aged~ aquarium
water, fresh synthetic sea water, and conditioned (aged)
synthetic sea water. This was done by adding 25-mL
portions to equal portions of each water type and
comparing to 25-mL portions of the untreated water type
mixed with 25 mL of deionized water in matched Nessler's
tubes (50 mL).
13~
-29-
The same kind of comparison was made using a ~5-ml sample
of a commercially available all purpose water conditioner.
There were no discernible differences between the
reference tubes and the test tubes in the experiments
with the different water types and the #l SFB solution.
This was also true of the tests with the all purpose water
conditioner instead of water. In the tests with the all
purpose water conditioner there were no discernible
differences after 21 days.
Example 8
Research was conducted to investigate the effects of SFB
solutions on seven different species of freshwater
fishes. The experimental design for these tests were
essentially the same throughout except for the number and
size of each species.
; 20 Two species were captured by seining and the other
five species were purchased from a tropical fish
wholesaler. The species chosen represented "typical"
families of ~reshwater a~uarium fishes and included
cyprinids, poeciliids, callichthyids, cichlids, characids,
centrarchids, and ictalurids. All fishes were
quarantined for a minimum of 14 days before being used in
any tests, and no species was used if any diseased or
dying individuals were observed among their population
until no disease or deaths had been observed for at least
10 days.
The test procedure for all species was the same. Three 4
liter beakers were filled with approximately 3,000 mL of
the water all from the same source. Aeration was provided
by a fine porosity airstone adjusted by valve so that
the fishes were not required to "fight" a current in the
beaker. ~ne of the three beakers was a control and had no
SFB solution addedO A second beaker had 0.4 mL of the #l
)()2~6
-30-
SFB solution added, which represented a double "normal"
dose of the active agent. A third beaker had 4.0 mL
added, which represented 10 times a "normal" dose.
Five of each species of cyprinid (gold barbs, Barbus
semi-fasciolatus), poeciliid (red-velvet swordtails,
Xipho~orus hell-erl), callichthyid (albino peppered
catfish, Corydoras paleatus), cichlid (silver angelfish,
Pterop~yllum emeeki), and characid (serpae tetra,
Hyphessobrycon callistus serpae) were used in each
beaker with one species per test. The cyprinids,
characids, and cichlids were tested for 24 and 48-hours
with two separate tests per species with different
individual Eishes used in each test. The other species
were each tested for 24 hours. The centrarchids
(longear sunfish, Lepomis megalotis) and ictalurids
(slender madtoms, Noturus ex _is) were used at the rate of
only one fish per beaker and then only the 24-hour series
were run for each species.
No deaths occurred with any of the species tested when SFB
solutions were added to aged aquarium water. This was
true for both the 24 and 48-hour tests.
Example 9
Research was conducted to investigate the effectiveness of
SFB for protecting fishes against toxic levels of chlorine
and chloramines.
Using municipal tap water, I tested the effectiveness of
the ~1 SFR solution for protecting silver angelfishes
against the toxic effects of chlorine and chloramines.
The same three-beaker set-up was used as in the previous
example, excep-t that unconditioned tap water was added
to the beakers after the water was allowed to flow from
the tap for a full ten minutes) Just as in the prior
tests, 5 angelfishes were added to each beaker. One
~3~286~
beaker served as a control with no SFB solution added.
The second and third beakers each received a 0.4 mL dose
of #l SFB solution. The fishes were observed during the
first four hours and then at the 8th hour and at the 24th
hour on termination of the test. The water was also
sampled for total and combined chlorine testing. Dead
fish were removed as found. The test procedure was
carried out twice in its entirety.
The municipal tap water utilized during the testing
was found to have a total chlorine content of 2.5 ppm and
a combined chlorine content of 2.0 ppm.
The experiment indicated that the #l SFB solution
protected the test fish, not only from death, but also
from the stressful effects of the new water. Within 1
hour, the fish in the control beakers having no SFB
solution had assumed stress coloration characterized by
darkening of their normal barred pattern. Within 2 hours,
40% of the fish in the control beakers were judged to
have lost the ability for normal swimming although they
still responded to a sharp tap on the side of the
beaker. At the conclusion of the tests, 50~ of -the fish
(3 out of 5 in test #1 and 2 out of 5 in test #2) in the
control beakers were dead. There were no mortalities
in the beakers to which SFB solutions had been added. At
1 hour, the beakers were all tested for total chlorine.
The control beakers showed no change in chlorine content
and the beakers containing SFB showed no chlorine
presence. There was still residual chlorine in the
control beakers at the conclusion of the tests after 24
hours.
Example 10
An investigation was undertaken to determine the effect,
if any, of overdose quantities of SFB on aquatic life in
marine watex.
-` ~L3~)028~
-32-
Thirty-six specimens of the pink-tipped anemone,
Condylactus ~iantea were distributed among four different
20-gal aquariums. The specific gravities of each tank
were adjusted so that the first tank was 1.016, the second
1.020, the third 1.025, and the fourth 1.030. The
anemones were allowed to acclimate to their tanks for 10
days. Ten-milliliter portions of the #l SFB solution were
pipetted, by bulb, onto the opened oral discs of three
anemones in each tank to observe the immediate reaction to
the solution in excess and the long-term reaction to
the solution in the tanks. Care was exercised to keep
from touching the animal with the tip of the pipette.
After the #l SFB solution was added, each tank was
observed for the first hour and then at 8 and 24 hours and
then ad lib. for the next three days.
I was unable to elicit any reaction from the anemones by
pipetting the #l SFB solution onto their oral discs.
However, in separate tests with deionized water
substituted for #1 SFB solution, all of the anemones
showed some reaction (6 oE 6) but only 2 of the animals
actually closed up in response to the test. No deaths
occurred among the population of test animals.
Example 11
Research was conducted to investigate the long-term
effectiveness of SFB for protecting fishes against toxic
levels of chloramines, chlorine and ammonia and the long-
term effect of overdose levels of sFs.
(1) Four, 20-gallon "community" aquariums were set up
with five silver angels, five gold barbs, six serpae
tetras, four red-velvet swordtails, and one albino
pepperred catfish per tank. Each tank was given
approximately a two-thirds water change daily, except
on the weekends. One tank (#7) was dosed with 5 mL of a
commercially available all-purpose water conditioner after
fresh tap water, without any -temperature adjustment, had
~3~2~
-33-
been added. A second tank (#8) was dosed with 5 mL of the
same all-purpose water conditioner before the tap water
was added. A third tank (#10) was dosed with 10 mL of the
#1 SFB solution for the Eirst 13 days of the test and
then, for the remaining 17 days, with 5 mL of #2 SFB
solution before the tap water was added. The fourth tank
($11) was dosed in the same manner as the third tank,
first with 10 mL of the #1 SFB solution and then with 5 mL
of #2 SFB solution after the tap water was added. This
test was conducted for a total of 30 days for a total
of 22 water changes on each aquarium.
(2) In eleven additional 20-gallon aquariums various
species and numbers were tested by adding 10.0 mL of the
#1 SFB solution each day, except weekends, for 23 days for
a total of 17 additions of #1 SFB solution and then by
adding 5 mL of #2 SFB solution each day, except weekends,
for 17 days for a total of 13 additions of #2 SFB. During
this 40-day period, no water was added or removed from the
eleven test aquariums.
The original aquarium census for these eleven aquariums
was as follows:
Tank #1 one 6" male bluegill sunfish (Lepomis macrochirus)
Tank #2 one 6" male longear sunfish
Tank #3 one 5" female longear sunfish ~ one 4" slender
madtom
Tank #4 fourteen 1" to 1-1/2" gold barbs
Tank #5 five 1" to 1-1/2" gold barbs ~ one 1" serpae tetra
Tank #6 five 1-1/2" silver angels
Tank #12 two 3" slender madtoms
Tank #13 one 7" green sunfish (Lepomis ~x~nellus)
Tank #14 ten 1" to 1-1/2" gold barbs
Tank #16 nine 1-1/2" silver angels
Tank #18 six 1-1/2" silver angels
The use, at first of 10 mL of the #l SFB solution, and
subsequently 5 mL of #2 SFB solution, resulted in exactly
~3002~
-34-
the same quantities of SFB being adcled to each of the test
aquariums. All of the test tanks were fed daily, ad lib,
except on weekends. All aquariums in these two tests were
filtered only with single under gravel filters. The
substrate in all test aquariums was l/~" x l/8" red
flint filter gravel at a depth of 3".
In the water changing tests with all-purpose water condi-
tioner and the SFB solutions, one fish (red-velvet
swordtail) died in tank #11 and one fish died in tank
#10 (red-velvet sword tail). In the all-purpose water
conditioner tanks, one silver angel died in tank #7.
These deaths were not considered significant. The general
health and appearance of all of the fishes were good. A11
of the fish were robust and ate with gusto when food
was ofEered.
In the daily-dosed tanks which had no water changes or
water additions, three (33-l/3~) angels died in tank #l.
There were no other deaths. All of the fishes ate
with gusto and were robust and healthy. All fishes
exhibited normal behavioral patterns such as begging~
displaying, schooling, and normal coloration.
Example 12
Research was conducted to investigate the effect of pH on
the reaction time of deamination in synthetic sea water
when using SFs solution.
In these tests, an ammonia specific-ion electrode (SIE)
and direct-reading meter were employed to track the change
in the relative free ammonia concentration over time. The
physical parameters for these tests include specific
gravity equal to 1.020 as determined by refractometer;
temperature equal to 20 +/- 1C; and total ammonia
concentration equal to l.00 ppm for one series of tests
and 5.00 ppm for a second series of tests. The tests were
~" ~3~2~
conducted in covered beakers containing 1 liter of the
test solution. The solutions were stirred continuously
during the tests. The ammonia SIE and meter were used in
a direct reading mode so that the electrode responded only
to the actual free ammonia in the solutions. For
those tests in which ~he total ammonia was 1.00 ppm, the
meter was given a "set concen-tration" equal to 1.00 and
5.00 for the 5.00 ppm tests. The meter then tracked the
actual, ~ree ammonia in the solution and displayed a
digital reading proportionate to the changes which
occurred over time as the reaction progressed.
Results of the effect on the neutralization reaction of
varying pH's in synthetic seawater and varying initial
concentration of ammonia are presented in Tables 5 -
11. This study reflects that the reaction favors an
increase in pH level and also favors higher initial
concentrations of ammonia.
Table 5.
pH = 6.000:
Relative Percent
Time: Conc. (ppm): Change:
0:00.00 1,00 0.00
O:Ol.00 0.993 0.79
0:02.00 0.992 0.80
0:05.00 0.985 1.50
0:10.00 0.974 2.60
0:20.00 0.953 4.70
0:30.00 0.935 6.50
1:00.00 0.889 11.10
2:00.00 0.825 17.50
4:00.00 0.728 27.20
5:00.00 0.712 28.80
6:00.00 0.683 31.70
13~)0~86;
-36-
Table 6.
pH = 7.000:
RelativePercent
Time: Conc. (ppm): Change:
0:00.00 1.00 0.00
0:01.00 0.976 2.40
0:02.00 0.951 4.00
0:05.00 0.898 lOo 20
0:10.00 0.878 12.20
0:20.00 0.781 21.90
0:30.00 0.715 28,50
1:00.00 0.578 ~.20
~:00.00 0.501 ~9.90
3:00.00 0.3~7 61.30
4:00.00 0.371 62.90
6:07.00 0.353 64.70
Table 7.
pH = 8.000:
, 25
RelativePercent
Time: Conc. (ppm): Change:
0:00.00 1~00 0.00
0 01.00 0.970 3.00
0:02.00 0.908 9.20
0O05.00 0.779 22.10
0:10.00 0.631 36.90
0:20.00 0.463 53.70
0:30.00 0.384 61.60
1:00.00 0.329 67.10
2:00.00 0.296 70.40
4:00.00 0.291 70.90
--` ~3~02~36
-37-
Table 8.
pH = 9.000:
Relative Percent
Time: Conc. (ppm): Change:
0:00.00 1.000.00
0:01.00 0.9158.50
0:02.00 0.83216.80
0:05.00 0.69130.90
0:10.00 ' 0.555 44.50
0:20.00 0.47752.30
0:30.00 0.44155.90
1:00.00 0.36763.30
2:05.00 0.29870.20
3:19~00 0.26873.20
Table 9.
pH = 6.000:
RelativePercent
Time: Conc. (ppm): Change o
2~ 0:00.00 5.00 0.00
0:01.00 4.96 0.80
0:02.00 4.93 1.40
0:05.00 4.80 4.00
0:10.00 4.69 6.20
0:20.00 4.41 11.80
1:00.00 3.90 22.00
2:00.00 3.46 30.80
3:00.00 2.94 41.20
4:00.00 2.61 47.80
5:00.00 2.55 49.00
6:00.00 2.41 51.80
6:40.00 2.39 52.20
7:00.00 2.32 53.60
-- ~.31)~28~
-38-
Table 10.
pH = 7.000:
Relative Percent
Time: Conc. (ppm): Change:
0:00.00 5.00 0.00
0:01.00 4.87 2.60
0:02.00 4.80 4~00
0:05.00 4.68 6.40
0:10.00 4.58 8.40
1:00.00 4.54 9.20
2:00.00 4.50 10.00
Table 11.
pH = 8.000:
Relative Percent
Time: Conc. (ppm): Change:
0:00.00 5.00 0.00
0:01.00 4.85 3.00
0:02.00 4.73 5.40
0:05.00 4.38 12.40
0:10.00 ~ 3.91 21.80
0:20.00 3.35 33.00
0:30.00 3.23 35.40
2:00.00 3.00 40.00
Example 13
Research was conducted to determine the eEfectiveness
of SFB solution to control losses of live marine animals
due to high total free ammonia levels as typically
encountered in shipping containers.
`
,
~30tl2~6
-39-
n these tests a variety of marine fishes and
invertebrates were collected in Nazareth Bay in St. Thomas
(U.S. Virgin Islands). These animals were packed in fresh
seawater treated with the #2 SFB solution dosed at the
rate of 4.93 mL/10 gallons of seawater in sealed
polyethylene bags. The bags were approximately 1.5 liter
in capacity. Each bag held approximately 500 mL of the
treated seawater along with the animals and 1 liter of
pure oxygen. Each bag was placed inside another bag and
sealed. The bags were placed in expanded styrene foam
boxes inside corrugated cardboard boxes. Four such boxes
containing a total of 65 bags of live animals were then
shipped from St. Thomas, via scheduled common air carrier,
to Miami, Florida, thence to Kansas City, Missouri. The
total shipping time was approximately 48 hours. No
untreated controls were used in this test.
It is well known among aquaculturists that marine fishes
and invertebrates suffer from and succumb to ammonia
build-up in shipping bags containing untreated water
and that significant losses can be expected. However,
there were no deaths of any of the marine fishes and
invertebrates shipped in ~2 SFB treated-seawater. The
health and condition of the fishes and invertebrates upon
arrival and subsequent removal to holding aquaria was
contrary to what would have occurred if no provision for
ammonia control had been made.
Example 14
A test was performed in which reduction in the measured
ammonia concentration was measured over time using 0.117
mL of the KFB solution added to 500 mL of a hard water
sample having a pH 8.0 at a temperature of 19.1 with a
total ammonia concentration of 1.00 ppm. The results
are shown in Table 12.
~L39~(~2~
-40-
Table 120
Time: Conc. (ppm):
0:00.00 1.00
0:01.00 0.892
0:02.00 0.837
0:05.00 0.759
0:10.00 0.722
0:20.00 0.683
A 31.7% reduction in the measured ammonia concentration,
~ at the end of 20 minutes, was demonstrated by the test.
1~
Example 15
Research was conducted to determine the effectiveness of
using a dry mixture form of SFB in place of SFB solutions
to neutralize aqueous free chlorine ~hypochlorites) and
ammonia.
A dry SFB mixture consisting of 58.995% sodium
formaldehydebisulfite and 41.005% fine blending salt
~sodium chloride) was used, volumetrically, at the same
rate as for #2 SFB solution (i.e., 1 teaspoon/10 gallons)
to treat two 20-gallon (75 liters) aquariums
containing 17 gallons (64 liters) each of aged
freshwater. The aged water had been pooled from water
taken from Eour aquariums in which a mixed population of
fishes had been maintained for 8 months. In the first
aquarium, a quantity of commercial bleach was added to
achieve a concentration of 2 ppm total chlorine. In the
second aquarium, a quantity of ammonium chloride solution
was added to achieve a concentration of 1 ppm total
ammonia. Into each aquarium, 8.4 mL of the dry SFB
mixture was measured by graduated cylinder and added
without mixing. Each aquarium was equipped with an air
diffuser to provide mixing and circulation of the water.
-" ~L3~02136
-41-
The pH and temperature of each aquarium was noted at the
beginning and end of the tests. The initial pH and final
pH remained the same at 6.8. The initial temperatures
were 21.2 and the final temperatures were 21~4 after 6
hours. No animals were maintained in the aquariums
during the tests.
Samples of 100 mL were drawn from each aquarium at the
start of the test and at time intervals of 1 hour, 2
hours, 3 hours and 6 hours for determination of the
concentrations of the target toxicants.
Table 13.
Chlorine Ammonia
Time: Conc. (Aq. #1): Conc. (Aq. #2):
0:00.00 2.0 ppm 1.00 ppm
1:00.00 0.00 ppm ~608 ppm
2:00.00 0.00 ppm .516 ppm
3:00.00 0.00 ppm o401 ppm
6:00.00 0.00 ppm .349 ppm
The results were consistent with and comparable to those
obtained with #2 SFB solution. Thus, dry or solution
formulations proved effective to neutralize the toxicants.
As indicated, this invention provides a one step method
for timely neutralizing chloramines, chlorine and ammonia
from saline and fresh waters Eor use in aquaculture.
It is nontoxic to fishes, aquatic invertebrates, marine
and freshwater algaes, and to aquatic plants. It does not
cloud the culture water or react with dissolved oxygen in
the culture water. It functions effectively throughout
the pH range of 6.0 to 9.0 of waters in which most
aquatic life is found. It can be combined with known
water conditioning chemicals and with known therapeutic
agents used in aquaculture.
~L30~
-42-
From the Eoregoing it will be seen that this inven-tion is
one well adapted to attain all the ends and objects
hereinabove set forth, together with the other advantages
which are obvious and which are inherent to the invention.
It will be understood that certain features and
~ subcombinations are of utility and may be employed without
'~ reference to other features and subcombinations. This is
contempla-ted by and is within the scope of the claims.
Since many possible embodiments may be made of the
invention without departing from the scope thereof, it is
understood that all matter herein set forth or shown in
the research examples is to be interpreted as illustrative
and not in a limiting sense.
