Application Of Ozone To Recirculating Aquaculture Systems

Brian L. Brazil
The Department of Wildlife and Fisheries Science,
Virginia Tech
Blacksburg, Virginia, 24060

Steven T. Summerfelt
The Conservation Fund’s Freshwater Institute
PO Box 1746, Shepherdstown, West Virginia, 25443

George S. Libey
The Department of Wildlife and Fisheries Science,
Virginia Tech
Blacksburg, Virginia, 24060

Brian/George: please spell check, I did not have time.

Introduction

Water in recirculating aquaculture systems is continuously filtered and reused to maintain a

suitable rearing environment. Increasing water usage raises the mass of fish that can be
produced on a limited water resource and allows for more control over the culture
environment. Also, because of intensive water reuse, systems that recirculate water have
effluents that are relatively low in volume, but contain proportionally higher nutrient and
organic levels, which can be treated for discharge more readilly (on a total mass basis) than
other aquaculture effluents.

Use of recirculating systems to achieve environmental control is not without draw backs.
Reusing the majority of water each day requires processes that remove both dissolved and
particulate matter to maintain water quality. Focus is often placed on ammonia removal
within a biofilter and on processes that remove settleable and filterable solids. However, as
production intensified and water reuse increases, the removal of nitrite, dissolved organics,
termed refractory organics, and colloidal solids left by conventional filtering becomes
increasingly important.

Insifying production in recirculating systems generally places more organic and nitrogenous
load on the biofilter, which can reduce their capacity to complete the two-steps serial
conversion of ammonia to nitrite (nitritification) and nitrite to nitrate (nitratification).
Because the increased loading increases competition for space and oxygen to the detriment of
the bacteria that convert nitrite into nitrate (nitratifiers), a net increase in nitrite concentration
across the biofilter and within the recirculating system as a whole results. These increased
nitrite levels of can be toxic to fish.

Refractory organics accumulate because they are not readily biodegradable due to their size
or chemical nature and because daily replacement of water is usually less than 15 percent.
Colloidal solids levels increase in systems with low water exchange because they are
generally not removed well by common settling and filtration units (Chen et al., 1994).
Elevated concentrations of refractory organics and colloidal solids may elicit physiological
responses that lead to restricted growth and eventually increased mortality rates (references).

Nitrite, refractory compounds, and colloidal organic materials that accumulate in

recirculating systems can be removed by increasing the daily water exchange rates or by
introducing oxidizing agents. To avoid high water exchange rates, some closed-system
culturists have chosen to apply ozone because of ozone’s attractive physio-chemical
characteristics. Traditionally, ozone has been used to remove pathogens from discharges
and/or water supplies of aquaculture systems (Owsley 1991; Cryer 1992). Ozone can be
added to recirculating systems to support water treatment by: (1) directly oxidizing nitrite to
nitrate, (2) breaking relatively non-biodegradable refractory organic compounds into smaller
and more biodegradable compounds, and (3) precipitating dissolved organic molecules and
microflocculating colloidal organic matter, which improves their removal via settling,
filtration or foam fractionation.

Physical Properties

Ozone is a metastable molecule of oxygen possessing three electrons rather than two. This
unstable and highly reactive species is produced by the reaction:

O + O2 ⇒ O3

which is initiated by a high energy field (Bablon et al. 1991). The standard free energy of
formation Go (1 atm) = 161.3 kJ / mol reveals that thermal activation is impossible;
although, heat is frequently used to decompose ozone (Bablon et al. 1991).

Rice et al. (1981) reported that in the atmosphere the half-life of ozone was approximately 12
hours making storage of ozone difficult. Ozone must be generated on site because the heat
generated during compression destroys any ozone present and because gas concentrations
70% or greater can spontaneously explode (Bablon et al. 1991). Additionally, when liquid
oxygen containing dissolved ozone evaporates, ozone separates creating a mixture which has
exploded at concentrations as low as 30% (Kinman 1972).

Generation
High energy sources such as cheminuclear sources, corona discharge, electrolytic processes,
and ultraviolet light (wavelengths less than 200 nm) are capable of exciting the electrons of
the oxygen molecule (Bablon et al. 1991). Generation of ozone in corona fields, a high
energy field established between two dielectric metals, is the most common method used to
produce large quantities of ozone (Bablon et al. 1991). When dried air or gaseous oxygen is
passed through the energy field, a portion of the diatomic oxygen molecules are excited to
create triatomic ozone (Rosenthal 1981; Bablon et al. 1991). Ultraviolet light generators are
less expensive to purchase but energy requirements can be 30 times greater than corona
generation. However, improvements in lamp technologies may reduce production costs
(Bablon et al. 1991).

Either air or pure oxygen can be used as a feed gas. However, 2 to 3 times more energy is
required to produce ozone at similar concentrations using air rather than purified oxygen
(Bablon et al. 1991). Additionally, the output concentration from a generator can be roughly
doubled by using pure oxygen rather than air, because pure oxygen contains up to 77% more
oxygen per unit volume than air (Rice and Netzer 1984). Corona discharge generators have
been reported to produce 6-8% ozone in the feed gas, but more commonly produce 2-3%
(Hudlicky 1990). The efficieny of generation depends upon the concentration of oxygen in
the feed gas and the percentage of ozone produced; it requires about 10 kwh of electricity to
produce 1 kg ozone at a concentration of 4-6% in an oxygen feed gas (Carlins and Clark,
1982).

Reliable and efficient generation of ozone requires that the feed gas has a dew point
temperature less than 65°C, and is free of particles and coalescible oil mists (Dimitriou
1990). These impurities foul the dielectrics within the corona discharge cells and react with
the ozone, which can reduce generator output. Experience at the Freshwater Institute
(Shepherdstown, West Virginia) revealed that hydrocarbon contamination of the liquid
oxygen supplied feed gas reduced ozone generation efficiency and output concentrations
(Bullock et al., 1996). Hydrocarbons can be retained during the distillation of liquid oxygen
and, on occasion, can result in concentrations greater than 20 ppm in industrial grade liquid
oxygen. Hospital grade oxygen (99.9% pure), although more expensive, will alleviate this
problem, extending efficient operational periods. Dimitriou (1990) suggested a limit of 5
ppm hydrocarbon in the oxygen feed gas.

Treatment Applications

The decision to use ozone should be made realizing it is not a panacea to all production
problems. It is important the culturist precisely identify the goals of ozone injection for
economically effective use. Though ozone is a nondiscriminating agent, ambient condition
determines its efficacy.

Organic Oxidation

The presence of dissolved organic compounds is readily identified by the yellow/brownish

coloration of culture water. The main coloring agents, humic acid, amino acids, carboxylic
acids, carbohydrates, etc., are collectively referred to as humic substances (Hirayama et al.
1988). The accumulation of dissolved organic compounds, solids and oxygen demanding
matter in recirculating systems is dependent on feed input, water exchange rate, solids
removal in the clarifier, and to a lesser degree biofiltration. Organic concentrations increase
in proportion daily and cumulative feed inputs (Hirayama et al. 1988). Easter (1992)
observed maximum CBOD, COD, and DOC levels approaching 25, 100, 40 mg/L,
respectively, in recirculating systems rearing hybrid striped bass. While exact sublethal and
lethal concentrations of dissolved organics have been not been identified; however, the
accumulation of dissolved organics has been implicated as a possible caused for reduced fish
growth rates and reduced nitrification efficiencies (Hirayama et al. 1988; Morrison and Piper
1988; Easter 1992; Nunely 1992; Bosworth 1994).

Ozone and its reaction by products are capable of oxidizing a great many organic substances
(Rice et al., 1981; Bablon et al., 1991). Ozone has been reported to effectively decrease the
accumulation of non-biodegradable organic compounds in recirculating systems (Rosenthal
and Otte, 1980). First order reaction rates describe the oxidation of organic substrates (M)
with ozone:

d [M] / dt = k [O3] [M]

where the rate limiting substance can be either ozone or the substrate (Bablon 1991). The
oxidation potential of M must be high for degradation to occur. The reactivity with ozone is
usually more pronounced than with any other oxidizing agent (Bablon 1991). Sites of initial
reaction are multiple bonds (C C, C=C, C O R, C=C X) or negatively charge atoms (N,
P, O, S, and nucleophilic carbons), therefore strong initial reactivities are anticipated for
molecules possessing OH, CH3, or OCH3 groups and weaker reactivities with NO2, CO2H or
CHO constituents (Richard and Brener 1984; Hudlicky 1990; Bablon 1991). A complete
explanation of organic substrate degradation is beyond the scope of this discussion,
however, recognize that the for mentioned functional groups construct the organic
compounds dissolved in culture water making them readily available for chemical oxidation
which will produce biologically assimilable molecules (Baily 1972; Masschelein 1992;
Bablon 1991).

Ozone addition also reduces the accumulation of TSS and COD within recirculating systems,
probably due to improved filtration resulting from ozone-induced microflocculation and
precipitation of dissolved organic compounds (Summerfelt et al. 1996).

Ammonia Removal

Reduction of ammonia in the biofilters within recirculating systems is a major concern in

recirculating systems because ammonia is a by-product of metabolism and because unionized
ammonia is extremely toxic to fish at low levels (Schreck and Moyle 1992). Ozone can
directly oxidize ammonia to nitrate:

4O3 + NH3 ⇒ NO3- + 4O2 + H3O+

However, the reaction rate constants for this process are extremely slow at pH levels below
9.3 (Richard and Brener 1984; Bablon et al. 1991). Conditions conducive to direct
ammonia oxidation are not typical in recirculating aquaculture systems, freshwater or marine,
which operate at a pH level between 6.8 and 8.4.

Even so, ozone addition may change the water chemistry within a recirculating system
sufficiently to improved ammonia removal within the biofilter. Autotrophs of primary
concern, Nitrosomonas sp. and Nitrobacter sp., are responsible for the nitrification process
(Wheaton et al. 1994):

NH4+ + 1.5 O2 ⇒ H2O + NO2- + 0.5 O2 ⇒ NO3-

In this aerobic process, Nitrosomona sp. oxidize ammonia to nitrite which is further oxidized
to nitrate by Nitrobacter sp. Ozone oxidation of non-biodegradable organic compounds
makes them more biodegradable, i.e., the oxidized organic molecules can be more readilly
assimilated by heterotrophic microorganisms. Additionally, ozonation improves the removal
of solids within recirculating systems through precipitation of dissolved organic matter and
microflocclation of colloidal solids (Summerfelt et al. 1996), which reduces the total organic
loading on the biofilter compared to an unozonated system. Research at Virginia Tech (B.
Brazil, unpublished data) on freshwater recirculating systems employing rotating biological
contactors (RBCs) showed ammonia removal efficiencies nearly 26% higher in systems
receiving ozonation than in non-ozonated systems. Sutterlin (1984) and Paller and Lewis
(198) also reported improved nitrification when ozone was added to recirculating systems.
The authors suggest that reducing the total load of organics on the biofilter may reduce the
growth of heterotrophic bacteria and allow for more growth of autotrophic nitrifying bacteria.
Collins et al. (1975) demonstrated that autotrophic bacteria had significantly lower growth
rates than heterotrophic bacteria, which is a disadvantage to autotrophic bacteria when
competing for space and oxygen with heterotophics. Limiting the nutrient base available to
heterotrophs may restrict their growth and allow autotrophic colonies to expand (Wheaton et
al 1994).

Nitrite Removal

Unlike the reaction rates of ammonia oxidation, direct ozone oxidation of nitrite to nitrate
proceeds readily:

NO2- + O3 ⇒ NO3- + 3O2

The reaction rate constant, 3.3 - 3.7 x 105 M-1 s-1 is largely pH independent (Bablon et al.
1991). Ozone stoichiometrically oxidizes nitrite to nitrate (Bablon et. al. 1991); therefore,
addition of ozone to recirculating systems is beneficial because it reduces nitrite levels
compared to systems that do not receive ozone (Rosenthal, 1980; Sutterlin et al., 1984,
Rosenthal and Kruner, 1985; Paller and Lewis 1988; Summerfelt et al. 1996). If on occasion
the nitratification in the biofilter is lost, ozonation of the system will help prevent the
accumulation of nitrite, which is a substantial benefit.

Because routine ozonation reduces the nitrite concentration going to the biofilter, over a long
period ozonation also reduces the quantity of nitratifying bacteria in the biofilter and, thus,
reduces the total nitrite removal capacity of the biofilter. If ever regular ozone addition is
interupted, nitrite rapidly accumulates within the recirculating systems, which can produce
serious fish health problems (S. Summerfelt, Freshwater Institute, unpublished data).

Disinfection

Inactivation of pathogens is a function of the residual ozone concentration, residence time,

pathogen load, temperature, pH, and alkalinity (Sproul 1975; Legeron 1984; Masschelein
1992). Ozone and by-product radicals react with substances prefrentually based on oxidation
potential and stoichiometric amounts of the substrates present (Bablon et al. 1991).
However, becuase there is so much more organic carbon present than typical amounts of
ozone added to recirculating systems, ozone’s affect on microbrial reductions in recirculating
systems is limited (Kinman 1972; Rosenthal 1981; Legeron 1984; Bullock et al. 1996).

Residual concentrations ranging from 0.5 to 4.0 mg/L are typically used to eradicated water
borne pathogens (Kinmam 1972). In pure water systems, residual concentrations between 0.4
and 0.01mg/L have proven effective in reducing E. coli and S. faecalis (Kinman 1972; Block
1982). Kinman (1972) demonstrated that in the pure water systems a contact time as little as
15 seconds was sufficient to destroy 100% of the microbes present. However, most systems
exert an ozone demand which increases the amount of ozone that must be added to sustain an
ozone residual for a given contact time. For example, secondary effluent contacted for 10
minutes with greater than 50 mg of ozone per liter reduced bacteria counts by 99%; the
increased ozone demand from dissolved organics accounted for the large amount of ozone
that must be added just to achieve a 2 log10 reduction (Kinman 1972). Increasing either
residual ozone concentration or contact time are required to produce significant disinfection
in situations with high levels of organic matter (Kinmam 1972; Block 1982; Bablon et al.
1991; Bullock et al. 1996). Complete bacterial reductions is necessary when treating water
for human consumption, but may not be required in the treatment of fish-culture water.

Seasonal variations in the pathogen load is often a characteristic of surface waters, rendering
the source unusable to certain aquaculture applications without disinfection (Conrad et al.
1975; Piper et al. 1982). Hatcheries using surface waters are particularly interested in
reducing pathogen loads to reduce mortalities and improve the growth and quality of fish.
Rosenlund (1975), rearing rainbow trout, reported that maintaining residual ozone
concentrations between 0.1 to 0.6 mg/L in the effluent of their contactor insured complete
removal of bacteria. Conrad (1975), artificially increasing the pathogen load of Flexibacter
columnaris, observed a 99% reduction in bacteria counts after ozonation. While complete
sterilization was not accomplished significant increases in survival rates from 60 to 96%
were observed. Conrad (1975) remarked that water low in sediment and organic matter
enhanced the bactericidal efficacy of the low level ozonation used. Seasonal disease
outbreaks of Ceratomyxa shasta at the Cowlitz hatchery (Washington) resulting in a 62.5%
mortality rate were significantly reduced to 1.4% through ozonation; the authors reported that
steelhead and cutthroat trout reared in water ozonated with a CT > 0.84 min.mg/L were free
of infection (Tipping 1988).

The reduction of opportunistic pathogens should be the role of ozonation in closed systems
with respect to disinfection. McClave (1992) working with marine systems reported that
fungal and yeast incidences were lower after ozonation than after chlorination. Collins
(1992) reported that attempts to achieve disinfection failed when the an ozone dose of 0.1 to
0.15 mg/L was applied to marine mammal exhibition pools. The author indicated that total
ozone demand of system required higher dosages to affect disinfection. In closed systems,
organic loads can reach levels that might make disinfection economically prohibitive;
therefore, achieving a reduction in pathogen loading or another improvements in water
quality should be the goal of ozonation.

Injection Regime

Pivotal to effective ozone use are the method and location of injection and amount of ozone
injected. Ozone treatments can be applied as a batch injection, as a series of daily injections,
or continuously supplied throughout the day. The method and location of injection, along
with the amount of ozone to be injected, should be chosen to meet the primary treatment
goals characteristic of a given recirculating system.

Method

Batch injection refers to using one continuous injection period to apply the ozone treatment.
Serial injection periods can be used to apply the ozone treatment over a greater portion of the
day injection ozone several periods throughout the day. With continuous injection, ozone can
be added to the system 24 hours per day. The decision to use one method over the other
depends on the culturists management strategy, which may be linked to the feeding schedule.

Introduction of feed initiates the degradation of water quality. Easter (1992) and Herbst
(1994) conducted diurnal studies of recirculating systems to evaluate the impact of feed
delivery on rearing conditions. Approximately 3 to 4 hours after the delivery of feed,
dissolved organics (Herbst 1994) and ammonia concentrations (Easter 1992) peaked.
Dissolved oxygen concentrations dropped to their lowest within minutes and required up to 2
hours to return to prefered levels. Based on these results, we believe that batch treatment can
be employed to improve water quality if no more than three moderate feed allotments are
presented per day. Based upon experiences at Virginia Tech, injection should begin just prior
to final feeding and extent for at least 3 hours (B. Brazil, unpublished data). Employing
serial or continuous injection periods can be used regardless of the number of feedings, but
these injection periods should be no shorter than 15 minutes.

Batch ozone injection produces larger fluctuations in water quality than serial or continuous
ozone injection would produce. Continuous ozone addition provides the least fluctuations in
water quality, especially if fish are fed more than 3-4 times per day. Also, providing a given
ozone dose based on feed loading (i.e., daily mass of ozone added is proportional to the daily
mass of feed added) with continuous ozone addition requires less ozone addition per unit
time than a batch or serial injection strategy. However, ozone added by batch is always
introduced when water quality has reached its worst cyclical condition within the system;
therefore, batch ozone addition may produce more water quality benefits per unit of ozone
added than are produced by either serial or continuous ozone addition.

Implimenting batch injection, if done manually, is less complicated and cheaper than
employing a serial injection regime. Implicit in serial injection is the fact that the ozone
generator will be cycled off and on. An automated controlled systems would be best suited to
operate this generation system. A central computer can be used to calculate and control
delivery of the proper amount of ozone to multiple culture systems.

Location

Because ozone can be used for many differentfunctions, the site of ozone injection can have a
large affect on how the recirculating system responds to the treatment. Arguements can be
made for injecting ozone in several locations within a recirculating system, particularly
adding ozone just before the biofilter versus generating and adding ozone within the oxygen
used to supersaturate the water just before the fish culture tank. Regardless of where ozone is
injected, however, there could be problems with residuals concentrations affecting either the
fish within the culture tank or the bacteria within the biofilter.

Direct ozonation of the rearing tank to achieve an ozone residual is not recommended.
Ozone is extremely toxic at low concentrations. Researchers have reported 96-hour LC50s of
9.3 and 80 µg / L for rainbow trout (Wedemeyer et al. 1979) and striped bass larvae (Hall et
al. 1981), respectively. Wedemeyer et al. (1979) reported partial gill pathology and reduced
feeding were observed at a residual concentration of 5 µg / L. Most recently, Bullock et al.
(1996) reported that ozone destroys gill lamellar epithelium which then produces ionic
imbalances that lead to death.

Adding ozone before the biofilter may allow ozone residual to enter the biofilter; this residual
would expend itself on the biosolids present. In this manner, the biofilter can be used to
shield the culture tank from potentially harmful ozone residuals. Research at Virginia Tech
indicates that measurable ozone residual levels entering the biofilters did not adversly affect
the biofilters performance (B. Brazil, unpublished data). However, in certain instances, it is
possible that an ozone residual could damage the micro-organisms within the biofilter, which
would be indicated by an increase in ammonia or nitrite concentrations.

Possibly the most economical way of adding large quantities of ozone to a recirculating
system is to generate and add ozone within the oxygen feed used to supersaturate the water
just before entering the fish culture tanks (Bullock et al. 1996; Summerfelt et al. 1996).
Adding ozone within the oxygen feed gas takes advantage of an oxygen supply and gas-
transfer unit that the fish already require. Adding ozone just before the fish culture tank also
allows the ozone to act upon and reduce the nitrite and fish pathogens just before the water
contacts the fish. Adding ozone just before the fish culture tank, however, has the drawback
of potentially exposing fish to ozone residuals, if the residuals were allowed to accumulate
within the culture tank (Bullock et al. 1996). Bullock et al. (1996) showed that the risk of
exposing fish to ozone was reduced when lower ozone loading rates were used and when an
ORP controller was used to prevent ozone accumulation within each culture tank.
Additonally, the risk of exposing fish to ozone can be reduced by using an ozone contact
chamber, to retain the water for several minutes, before passing the water to the fish culture
tank. Others have reported on destruction of ozone residuals in water through the use of 254
nanometer ultraviolet-light (Cryer 1992).

As will be discussed later, ozonation can increase biogradeability and nitrification rates (see
the Application and Treatment section for more details). Yet, it is unknown whether-or-not it
is necessary to add ozone directly before the biofilter to positively affect nitrification within
the unit. However, it is likely that the point of ozone injection within the recirculating system
does not have a large affect on oxidation of non-biodegradable organic compounds and their
subsequent assimilation within the biofilter. Additionally, ozone will oxidize dissolved
organic compounds and colloidal solids within the water regardless of where ozone is
injected; However, microflocculation and precipitation of the destabilized compounds is
affected by the length of contact time before the solids removal device. The culture tank can
double for the chamber to provide contact time for microflocculation and precipitation.

Amount

Conrad et al. (1975), Tipping (1988), Blogoslawski (1992), Rueter and Johnson (1995) have
suggested treatment regimes to remove fish and crustaceans pathenogentic organism from
influent culture waters based on the CT concept (Legeron 1982). There has been little past
published work to suggest appropriate dosage rates for the reduction of organics in
recirculating aquaculture systems. Ozonation rates in published works (Williams 1982;
Sutterlin et al. 1984; Morrison and Piper 1988; Poston and Williams 1988; Poston and
Williams 1990) appear to be based on generator production capacities to evaluate ozone
treatment or treatment regimes used in the water industry. The authors believe that unless
disinfection is the objective of ozone injection, ozone should be injected to reduce organic
loads and increase biogradeability as is the case in wastewater treatment.

Research in a water recirculating system used to culture rainbow trout at the Freshwater
Institute indicated that an ozone dosing rate of 25 g of ozone per kilogram of feed improved
water quality and microscreen filtration (Summerfelt et al. 1996) and reduced bacterial gill
disease (BGD) associated mortalities and chemical treatments required to control BGD
epizootics (Bullock et al. 1996); adding ozone at a higher rate 36-39 g of ozone per kilogram
of feed had roughly the same affect on water quality, microscreen filtration, and BGD
epizootics, but was much more likely to produce fish mortality when on occasion ozone
accumulated to toxic levels (Bullock et al. 1996). Research at the Freshwater Institute also
indicated that neither ozone dosing rate was sufficient to produce greater than even a 1 log10
reduction in the numbers of heterotrophic bacteria in the system water or on gill tissue
(Bullock et al. 1996). Rapid loss of oxidation capacity (ozone half-life < 1-15 sec) caused by
levels of nitrite and organic carbon was blamed for the failure of ozone to reduce numbers of
heterotrophic bacteria.

Brazil (1996) reported that between 10 and 15 g of ozone of per kilogram of feed delivered
per day should provide an adequate reduction of dissolved organic concentrations. Rearing
hybrid striped bass, Brazil (1996) found that DOC concentrations did not differ significantly
when compared to levels measured in systems not receiving ozone injection; however, the
ratio of DOC: accumulated feed input differed significantly. Increased growth rates and
lowered turbities were positively correlated to ozone dosage rate. However, diminishing
returns were discovered for growth rates at dosage rates greater than 13g of ozone per
kilogram of feed. Research at Virginia Tech (B. Brazil, unpublished data) does indicate that
a minimum ozone dosage should be no lower than 5 - 7 g of ozone per kilogram of feed
(???in conjuction with foam fractionation????).

Brian/George: discussing foam fractionation here is out of place because it has not been
discussed previously.

Foam fractionation reduces the dissolved organics concentration thereby lowering the ozone
demand of the system (Weeks and et al. 1992; Chen et al. 1993)

Table 1. Definition of some terminology commonly used to describe ozonation.

(Brian/George: some of these definitions need work and please place this table were you
think it would fit best.)
Parameter Definition
Applied dosage rate amount of ozone injected per treatment basis per injection period (mg / L /
unit of time)
C.T a measure of microbial reduction potential based upon the product of
the ozone residual concentration times the contact time
Dosage rate amount of ozone injected per treatment basis [M] (g / M)
Injection period the length of time ozone is injected (time)
Injection rate amount of ozone in the gas stream supplied to the contactor per hour
(mg / L / h, g / m3 / h)
Ozone concentration the amount of ozone present within the gas stream (% ozone by weight, mg /
L)
Ozone production the amount of ozone generated daily (g / day)
Ozone residual concentration of measurable ozone leaving the contactor (mg / L)
Ozone treatment total amount of ozone to be injected daily (g / day) = dosage rate * M / day

Summary

The implementation of an ozonation regime can only be accomplished knowing the

goals of the treatment. The culturist must understand the limitations of ozone and impact that
it will have on system performance. Before ozone is injected it is recommended that other
literature be reviewed. This discussion was intended to be a starting point to suggest
injection strategies and dispell a few myths about ozone. Ozone can be of great benefit to
increasing production, but it can also be even greater detriment if not used properly.

Concepts paramount to economically efficient ozonation in closed systems are unit process
optimization and diminishing turns. Optimization is accomplished by sizing the process
according to the total waste produced by the introduction of feed (Lucchetti and Gray 1988),
ozone dosage rates should be no exception. Applying too little or too much ozone would be
economically ineffecient. The difficult part is determining the optimum ozone dosage rate.
References

Bablon, G., W.D. Bellamy, M. Bourbigot, F.B. Daniel, M. D., F. Erb, B. Gordon, B.
Langlais, A. Laplanche, B. Legube, G. Martin, W. J. Masschelein, G. Pacey, D.A. Rechhow
and C. Ventresque. 1991. Fundamental Aspects. Pages 11-132 in Bruno Langlais, David A.
Reckhow and Deborah R. Brink, editors. Ozone in Water Treatment: Application and
Engineering. AWWA Research Foundation (Denver, Colorado).

Bailey, P.S. 1972. Organic groupings reactive toward ozone mechanisms in aqueous media.
Pages 29-60 in F.L. Evans III. Ozone in water and wastewater treatment. Ann Arbor Science
Publishers Inc. Ann Arbor, Michigan.

Block, J.C. 1982. Removal of bacteria and viruses by ozonation. Pages 66-68 in W.J.
Masschelein. Ozonization manual for water and wastewater treatment. John Wiley and
Sons, New York.

Blogoslawski, W.J. 1992. Ozone treatment of seawater to control vibriosis in mariculture of

penaid shrimp, Penaeus vanameii. Abstracts 3rd International Symposium On The Uses of
Ozone In Aquatic Systems. The International Ozone Association, Pan American Committee
and The New York Aquarium, Osborn Laboratories of Marie Sciences, Greenwich,
Connecticut

Bosworth, B.G. “Use of Species genetic resources in Morone breeding programs.” Ph.D.
Dissertation, Virginia Polytechnic Institute and State University. 1994.

Brazil, B.L. “Impact of Ozonation on System Performance and Growth Characteristics of

Hybrid Striped Bass (Morone chrysops X M. saxatilis) and Tilapia Hybrids (Sarotherodon
sp.) Reared in Recirculating Aquaculture Systems.” Ph.D. Dissertation, Virginia Polytechnic
Institute and State University. 1996.

Bullock, G. L., Summerfelt, S. T., Noble, A., Weber, A., Durant, M. D., and Hankins, J. A.
“Effects of ozone on outbreaks of bacterial gill disease and numbers of heterotrophic bacteria
in a trout culture recycle system.” In Successes and Failures in Commercial Recirculating
Aquaculture (Conference Proceedings). NRAES-98. Northeast Regional Agricultural
Engineering Service: Ithaca, New York. July 1996.

Carlins, J. J., and R. G. Clark. “Ozone generation by corona discharge.” Pages 41-76 In R. G.
Rice and A. Netzer, editors. Handbook of Ozone Technology and Applications, Volume 1.
Ann Arbor Science Publishers. Ann Arbor, Michigan. 1982.

Chen, S., M.B. Timmons, J.J. Bisogni, Jr., and D.J. Aneshansley. “Protein and its removal
by foam fractionation.” The Progressive Fish-Culturist 55(1993) 76-82.
Collins, M.T., J.B. Gratzek, E.B. Shotts Jr., D.L. Dawe, L.M. Campbell, and D.R. Senn.
1975. Nitrification in an aquatic recirculating aquaculture system. Journal of Fisheries
Research Board of Canada 32:2025-2031.

Conrad, J.F., R.A Holt, and T.D. Kreps. 1975. Ozone disinfection of flowing water. The
Progressive Fish-Culturist 37:134-136.

Cryer, E. “Recent applications of ozone in freshwater fish hatchery systems.” Pages 134-154
In W. J. Blogoslawski, editor. Proceedings of the 3rd International Symposium on the Use of
Ozone in Aquatic Systems. International Ozone Association, Pan American Committee.
Norwalk, Connecticut. 1992.

Easter, C. 1992. Water chemistry characterization and component performance of a

recirculating aquaculture system producing hybrid striped bass. M.S. Thesis, Virginia
Polytechnic Institute and State University.

Hall, L.W. Jr., D.T. Burton, and L.B. Richardson. “Comparison of ozone and chlorine
toxicity to the developmental stages of striped bass, Morone saxatalis.” Canadian Journal of
Fisheries and Aquatic Sciences 38(1981):752-757.

Hirayama, K., H. Mizuma, and Y. Mizue. 1988. The accumulation of dissolved organic
substances in closed recirculation culture systems. Aquacultural Engineering 7:73-87.

Hudlicky, M. 1990. Oxidations in organic chemistry. American Chemical Society. Page 4.

Kinman, R.N. 1972. Ozone in water disinfection. Pages 123-143 in F.L. Evans III. Ozone
in water and wastewater treatment. Ann Arbor Science Publishers Inc. Ann Arbor, Michigan.

Lawson, T.B. 1995. Fundamentals of aquacultural engineering. Chapman and Hill , New
York, New York.

Legeron, J.P. “Contact time of ozonation.” Ozonization Manual for Water and Wastewater
Treatment. W.J. Masschelein, editor. John Wiley and Sons, New York. 1982. Pages 133-
136.

Legeron, J.P. 1984. Ozone disinfection of drinking water. Pages 1-20 in R. G. Rice and A.
Netzer, editors. Handbook of ozone technology and application volume II: ozone for
drinking water treatment. Butterworth Publishers, Boston, Massachusetts.

Lucchetti, G.L and G.A. Gray. “Water reuse systems: A review of principal components.”
The Progressive Fish-Culturist 50(1988):1-6.

Masschelein, W.J. 1992. Unit processes in drinking water treatment. Marcel Dekker, Inc.
New York.
McClave, C.A. 1992. A comparative study of microbial contents of a chlorinated vs an
ozonated/chlorinated marine mammal system. 3rd International Symposium On The Use of
Ozone In Aquatic Systems. The International Ozone Association, Pan American Committee
and The New York Aquarium, Osborn Laboratories of Marine Sciences. Greenwich,
Connecticut.

Metcalf and Eddy, Inc. 1991. Wastewater engineering: treatment disposal, and reuse. 3rd
edition. McGraw Hill Inc., New York, New York. Pages 70-77.

Morrison, J.K. and R.G. Piper. 1988. Effect of Reused water on atlantic salmon. The
Progressive Fish-Culturist 50:110-112.

Nunely, C.E. 1992. Production of hybrid striped bass (Morone chrysops X Morone
saxatilis) in a recirculating aquaculture system. M.S. Thesis, Virginia Polytechnic Institute
and State University.

Owsley, D. E. “Ozone for disinfecting hatchery rearing water.” Pages 417-420 In J. Colt and
R. J. White, editors. American Fisheries Society Symposium 10. American Fisheries Society,
Bethesda, Maryland. 1991.

Paller, M.H. and W.M. Lewis. 1988. Use of ozone and fluidized-bed biofilters for increased
ammonia removal and fish loading rates. The Progressive Fish-Culturist 50:141-147.

Piper, R.G., I.B. McElwain, L.E. Orme, J.P McCraren, L.G. Fowler, and J.R Leonard. 1982.
Fish hatchery management. United States Department of the Interior, Fish and Wildlife
Service, Washington, D.C.

Poston, H.A. and R.C. Williams. “Interrelations of Oxygen Concentration, Fish Density, and
Performance of Atlantic Salmon in an Ozonated Water Reuse System. The Progressive Fish-
Culturist 50(1988):69-76.

Poston, H.A. and R.C. Williams. “Effects of Ozonated-Water Reuse on Salinity Tolerance of
Altanic Salmon.” The Progressive Fish-Culturist 52(1990):36-40.

Rice, R.G., C.M. Robson, G.W. Miller, and A.G. Hill. 1981. Uses of ozone in drinking
water treatment. Journal of American Water Works Association 73:1-44.

Richard, Y. And L. Brener. 1984. Removal of ammonia and other nitrogen derivatives from
drinking water with ozone. Pages 99-140 in R. G. Rice and A. Netzer, editors. Handbook of
ozone technology and application volume II: ozone for drinking water treatment.
Butterworth Publishers, Boston, Massachusetts.

Rosenthal, H., 1981. “Ozonation and sterilization.” Pages 219-274 In K. Tiews, Editor.
Proceedings of World Symposium on Aquaculture in Heated Effluents and Recirculation
Systems, Berlin. pp. 219-274.
Rosenthal, H. and Kruner, G. “Treatment efficiency of an improved ozonation unit applied to
fish culture situations.” Ozone: Sci. Eng., 7(1985): 179-190.

Rosenlund, B.D. 1975. Disinfection of hatchery influent by ozonation and the effects of
ozonated water on rainbow trout. Pages 59-69 in W.J Blogoslawski and R.G. Rice, editors.
Aquatic applications of ozone. The International Ozone Institute, Inc., Syracuse, New York.

Rueter, J. and R. Johnson. 1995. The use of ozone to improve solids removal during
disinfection. Aquacultural Engineering 14:123-141.

Sproul, O.J. 1975. Virus inactivation by ozone. Pages 16-24 in W.J Blogoslawski and R.G.
Rice, editors. Aquatic applications of ozone. The International Ozone Institute, Inc.,
Syracuse, New York.

Standard Methods for the Examination of Water and Wastewater. 1992. 17th edition (1989)
Edited by Clesceri, Greedberg, and Trussell. American Public Health Association.
Washington D.C.

Summerfelt, S. T. “Effects of ozone on microscreen filtration and water quality

in a recirculating rainbow trout culture system.” In Successes and Failures in Commercial
Recirculating Aquaculture (Conference Proceedings). NRAES-98. Northeast Regional
Agricultural Engineering Service: Ithaca, New York. July 1996.

Sutterlin, A.M, C.Y. Couturier, and T. Devereaux. 1984. A recirculating system using ozone
for the culture of Atlantic salmon. The Progressive Fish-Culturist 46:239-244.

Tipping, J.M. 1988. Ozone control of ceratomyxosis: survival and growth benefits to
steelhead and cutthroat trout. The Progressive Fish-Culturist 50:202-210.

Uhlig, G. 1984. Closed system for generating ozone from oxygen and its application to the
treatment of drinking water at Duisburg, Federal Republic of Germany. Pages 141-173. In
Rip G. Rice and Aharan editors. Handbook of ozone technology and applications volume II:
ozone for drinking water treatment. Butterworth Publishers, Boston, Massachusetts.

Wedemeyer, G.A., N.C. Nelson, and W.T. Yasutake. “Physiological and biochemical aspects
of ozone toxicity to rainbow trout (Salmo gairdneri).” Journal Fisheries Research Board of
Canada 36(1979):605-614.

Weeks, N.C., M.B. Timmons, and S. Chen. “Feasibility of using foam fractionation for the
removal of dissolved and suspended solids from fish culture water.” Aquaculture
Engineering 11(1992) 251-265.

Wheaton, F.W. 1977. Aquacultural Engineering. John Wiley and Sons, New York.
Wheaton, F.W., J.N. Hocheimer, G.E. Kaiser, M.J. Krones, G.S. Libey and C.C. Easter.
1994. Nitrification filter principles. Pages 101-171. In M. B. Timmons and T.M. Losordo,
editors. Developments in Aquaculture and Fisheries Science, volume 27: Aquaculture Water
Reuse Systems: Engineering Design and management. Elsevier Science B.V. Amsterdam,
The Netherlands.

Williams. R.C., S.G. Hughes, and G.L. Rumsey. 1982. Use of ozone in a water reuse system
for salmonids. The Progressive Fish-Culturist 44:102-105.