Reviews in Aquaculture, 1–31 doi: 10.1111/raq.

12354

Nitrite implications and its management strategies in

aquaculture: a review
Alexander Ciji and Mohammad Shahbaz Akhtar
ICAR-Directorate of Coldwater Fisheries Research, Bhimtal, Nainital, India

Correspondence
Dr Mohammad Shahbaz Akhtar, Nutritional
Physiology Lab., ICAR-Directorate of
Coldwater Fisheries Research, Bhimtal-
263136, Nainital, Uttarakhand, India. E-mail:
mdshahbazakhtar@gmail.com
Received 31 August 2018; accepted 11 May
2019.
Abstract
High nitrite levels are often encountered in intensive and recirculatory aquacul-
ture systems but can also be found in natural waters under certain circumstances.
Significant research advancement on nitrite and its implications in aquaculture
sector has been made for last few decades, but emphasis on its management
aspects has been given due consideration since recent past. Therefore, this review
article not only aims to summarize the voluminous published reports on nitrite
implications in fish but also recapitulates the research progress made on mitiga-
tion/management strategies. Nitrite exposure has been shown to adversely affect
fish and shellfish growth, moulting, blood oxygen carrying capacity, water bal-
ance, osmoregulation, ionic homoeostasis and cause endocrine disruption. The
nitric oxide produced from nitrite can inhibit several components of steroido-
genic pathway leading to reduced reproductive performance. Different
approaches and management strategies that have been evolved so far to address
the nitrite-associated stress have varying degrees of success. Adopting better nutri-
tional approach like feeding antioxidant or nutraceutical fortified feeds along with
effectively functioning biological filters is found to be helpful in reducing the dele-
terious effects of nitrite in cultured animals. This review paper, containing the
plethora of information on nitrite and its management in one place, will definitely
act as a ready reference for different stakeholders such as farmers, researchers and
policymakers for sustainable development of aquaculture.
Key words: aquaculture, aquaponics, methaemoglobin, nitrite, nutritional intervention, salt
addition.

bacteria (NOB) (Nitrobacter spp., Nitrospira spp.).

Introduction
The world aquaculture production will need to reach
102 million tons by 2025 to sustain the present level of
consumption (FAO 2016). Aquaculture intensification is
one of the potential strategies to meet this projected
requirement as land resources are shrinking for its
expansion. However, the excessive use of proteinaceous
feed, nitrogenous fertilizers along with higher fish stock-
ing densities in intensified aquaculture result in increased
load of nitrogenous as well as other toxic metabolites in
the waterbodies (Das et al. 2004a). Frequent aeration or
strong wind action can also raise nitrite levels because of
increased pond mixing of nitrites produced in oxygen-
deficient bottom muds. During bacterial nitrification,
ammonia is firstly oxidized to nitrite by ammonia-oxi-
dizing bacteria (AOB) (Nitrosomonas spp. or Nitrosospira
spp.) and then to less toxic nitrate via nitrite-oxidizing
Hence, nitrite remains available in water as an interme-
diate in the bacterial oxidation of ammonia, the nitroge-
nous excretory product of fish, to nitrate. Hence,
imbalances in bacterial nitrification process can also ele-
vate the nitrite concentrations in water (Deane & Woo
2007). Based on thermodynamics and electron transfer,
the ratio of AOB to NOB in a balanced nitrifying system
should be theoretically 2:1 (Hooper et al. 1997). How-
ever, controversial disproportionate AOB/NOB ratio has
also been reported by other investigators (Mari et al.
2012; Ramdhani et al. 2013). The bacteria Nitrosomonas
spp., which converts ammonia to nitrite, functions well
at temperatures of 15–20°C. However, Nitrobacter spp.
the bacteria that transform nitrite to nitrate function
poorly at these temperatures and prefer a pH range of
7.5–8.6 with at least 1 mg L
1 dissolved oxygen. There-
fore, increased nitrite toxicity is reported in conditions

© 2019 Wiley Publishing Asia Pty Ltd 1

A. Ciji and M. S. Akhtar
of lower pH and cool temperature in several teleosts and
crustaceans (Huey et al. 1984; Chen & Cheng 2000).
In traditional extensive aquaculture systems, the nitrite
levels seldom exceed the tolerance/desirable limit of
1 mg L
1 to fish (<0.1 mg L
1 Boyd & Tucker 1998 and
0.005–0.5 mg L
1 Stone & Thomforde 2004). Nitrite
implications are more likely to occur in closed intensive
culture systems due to insufficient, inefficient or malfunc-
tioning filtration systems for removing waste ammonia
from water through nitrification (Kroupova et al. 2005).
Nitrite build up in intensive aquaculture systems can
exceed 50 mg L
1 (Kamstra et al. 1997). Furthermore, the
limited capacity of water resources urges the need of recir-
culating aquaculture systems. In recirculatory aquaculture
systems, the ultraviolet irradiation used for water disinfec-
tion may be another source of nitrite if the treated water
contains high nitrate levels, as nitrate can be converted to
nitrite by UV irradiation (Kroupova et al. 2018; Lu et al.
2009; Sharrer et al. 2005). Elevated nitrite concentration
may also occur as a result of changes in the pattern of agri-
cultural practices, food processing and industrialization
(Chow & Hong 2002). It has been reported that the con-
centration of nitrite increases as high as 20 mg L
1 in
shrimp ponds (Tacon et al. 2002) and causes high mortali-
ties (Guo et al. 2013). In nutshell, the four primary sources
of nitrogenous wastes in aquaculture are: the excretory
products of fish, organic debris from dead and dying
organisms, uneaten feed and faeces and nitrogen gas from
the atmosphere.
Nitrite also plays an important role in the nitrous oxide
(N2O) emission from aquaculture systems. Hu et al. (2012)
reviewed that aquaculture can be an important anthro-
pogenic source of the greenhouse gas nitrous oxide which
has a global warming potential 310 times that of carbon
dioxide over a 100-year lifespan. During microbial nitrifica-
tion and denitrification process, N2O is produced and is
diffused into the atmosphere. The average N2O emission
factor of aquaculture system is calculated to be 1.69 g
N2O
N per kg fish produced (Hu et al., 2012). Consider-
ing the present growth rate of aquaculture, the global
N2O
N emission from aquaculture in 2030 is estimated
to be 3.83 9 1011 g by Hu et al. (2012) and 1.01 9 1012 g
by Williams and Crutzen (2010). Hence, nitrite is not only
a potential foe of aquaculture industry but also a major
challenger to the global environment.
This elevated nitrite is a potential factor eliciting stress in
aquatic animals (Palachek & Tomasso 1984a,1984b; Sam-
paio et al. 2002) resulting disease outbreaks and enormous
losses (Kshirsagar et al. 1995; Maag & Vinther 1996; Ole
et al. 2009). The acute and chronic effects of nitrite on dif-
ferent aquatic organisms have extensively been reviewed by
several authors (Philips et al. 2002; Jensen 2003; Camargo
et al. 2005; Kroupova et al. 2005; Camargo & Alonso 2006;
2 © 2019 Wiley Publishing Asia Pty Ltd
Tomasso 2012; Ramano & Zeng 2013; Kroupova et al.
2018). However, very less emphasis has been given to its
remedial measures which are important for aquaculture
management and sustainable production. Additionally,
most of the studies and reviews focussed on the effects of
nitrite on haematologic and metabolic alterations as well as
effects on ion exchange process while limited attentions
were given to endocrine disruptions. In the present review,
not only a comprehensive account on physiological conse-
quences of nitrite, the potential mitigational/management
strategies to be adopted for minimizing the deleterious
effect of nitrite exposure are also elaborately discussed.
Symptoms of nitrite toxicity in fish
Toxicity of nitrite to fish varies greatly depending on a vari-
ety of external and internal factors. Some fish may simply
be lethargic hovering at the bottom of the tank, while
others may die suddenly with no apparent signs of infec-
tion. Usually, gross lesions are not apparent. Common
symptoms of higher levels of nitrite include gasping at the
surface of the water, hanging near water outlets, increased
heart rate, hyperventilation, swollen, flared and brown-
coloured gills. Furthermore, the colour of freshly drawn
blood may be light brown. Sometimes, the skin may appear
reddish with excessive mucus production. Even with opti-
mum dissolved oxygen levels of the rearing water, the
affected fish may frequently show signs of hypoxia. Hyper-
plasia, vacuolization and elevated numbers of chloride cells
were noticed in the gills of nitrite-affected carps (Svo-
bodova et al. 2005a). The early recognization of nitrite
intoxication through these symptoms could be useful to
prevent severe impairment and loss of the cultured fish.
Nitrite uptake and accumulation
Aquatic animals including fish are more susceptible to high
nitrite concentrations than terrestrial animals. Nitrite
enters the fish mainly by diffusion through the skin and
gills. It is highly problematic in freshwater fish, as they are
hyperosmotic to their environment. Active ions uptake
across the gills is required for freshwater fish to maintain
ionic homoeostasis (Kroupova et al. 2005). They absorb
nitrite principally through the chloride cells of the gill by
competing with active chloride uptake and can accumulate
to high levels in the blood (Kroupova et al. 2005). There-
fore, nitrite intoxication is more prevalent in fish with
higher branchial chloride uptake rates (Williams & Eddy
1986). Additionally, other tissues such as gill, brain, liver
and muscle also can accumulate significant amounts of
nitrite (Margiocco et al. 1983). Long-term exposure to even
low concentrations of nitrite can damage the immune sys-
tem of fish and thereby make them more vulnerable to
Reviews in Aquaculture, 1–31

secondary diseases. However, in marine fish, the nitrite is
less toxic as nitrite uptake through gills is against the active
chloride transport (Camargo et al. 2005). However, studies
in several marine species revealed that elevated nitrite levels
reduced growth (Siikavuopio & Sæther 2006), increased
methaemoglobin formation (Scarano & Saroglia 1984),
caused osmoregulatory disorders (Deane & Woo 2007) and
altered normal physiology (Grosell & Jensen 2000; Park
et al. 2013). Several studies described a positive correlation
between nitrite concentration in blood and methaemoglo-
bin formation in salmonids, ictalurids and cyprinids (Bath
& Eddy 1980; Huey & Beitinger 1982). In marine fish,
nitrite uptake possibly occurs across the intestinal and gill
epithelium (Grosell & Jensen 2000; Jensen 2003; Deane &
Woo 2007; Tomasso 2012).
Physiological consequences of nitrite toxicity
Nitrite has been recognized as a toxic compound and can
induce a variety of physiological alterations in fish and the
toxicity may result from a combination of physiological/
biochemical effects. The toxicity varies with the speed and
degree of nitrite accumulation, environmental conditions,
duration of exposure and species as well as on the endoge-
nous capacity for detoxification and elimination of nitrite.
Nitrite exposure has been shown to adversely affect fish
growth (Woo & Chiu 1994), gill osmoregulatory processes
(Woo & Chiu 1994), blood oxygen-carrying capacity (Jen-
sen 2003), ionic homoeostasis (Grosell & Jensen 2000; Jen-
sen 2003) and more recently reported to cause endocrine
disruption (Edwards & Guillette 2007). Various physiologi-
cal disturbances induced by high nitrite are described here-
under:
Reduced growth and feed intake
A reduction in feed intake was noticed in nitrite exposed
fish and shrimp (Campos et al. 2013; Roques et al. 2013).
Nitrite exposure has been shown to adversely affect growth
in several fish and shellfish species (Colt et al. 1981; Frances
et al. 1998; Koo et al. 2005; Mallasen & Valenti 2006;
Siikavuopio & Saether 2006; Ciji et al. 2013a). This decrease
in growth rate can be due to reallocation of energy towards
maintenance and repair during stress (Kumaraguru & Bea-
mesh 1986). Furthermore, the increased energy demand for
the detoxification process of nitrite may also result in
growth retardation. The increased energy expenditure for
osmoregulation and ionic homoeostasis (Chen & Cheng
1993; Cheng et al. 2013) along with reduced feed intake
may be attributed to reduced growth observed in nitrite-ex-
posed fish. Furthermore, Han et al. (2017) reported alter-
ations in digestive enzyme synthesis due to decreased
number of R cells (the major site of digestive enzyme
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
synthesis) in the hepatopancreas of Litopenaeus vannamei
at higher ammonia and nitrite which resulted in reduced
metabolic energy hampering its growth. Nitrite exposure
also increased the moulting frequency in Penaeus monodon
juveniles with less size increment at each moult leading to
slow growth rate (Chen & Chen 1992). Besides, Portunus
pelagicus showed significantly increased ‘moult death syn-
drome’ characterized by the inability to completely shed
the old exoskeleton at higher nitrite levels (Romano & Zeng
2009). Furthermore, the higher oxygen consumption dur-
ing moulting and consequent greater intoxication with
nitrite have been suggested for the higher mortality rate
observed during moulting period in nitrite exposed
shrimps (Wasielesky 2000; Furtado et al. 2016).
Methaemoglobinaemia and other haematological
alterations
Blood is the primary target site of nitrite action in fish, and
the major physiological effect of nitrite is the conversion of
haemoglobin to methaemoglobin. Nitrite oxidizes iron in
haemoglobin from 2+ to 3+ oxidation state leading to the
formation of methaemoglobin or ferrihaemoglobin and this
physiological condition is known as methaemoglobinaemia
(Kiese 1974; Jensen 2003; Kroupova et al. 2005). The
methaemoglobin is not able to transport oxygen to tissues
leading to tissue hypoxia (Cameron 1971; Kroupova et al.
2005), and therefore, the affected fish gasp at the water sur-
face even if the oxygen level in the rearing water is relatively
high. Respiratory system is the chief target of the toxic effect
of higher nitrite. The higher proportion of methaemoglobin
gives blood a brownish colour (Cameron 1971) and hence
also known as brown blood disease. Studies described that
fish could withstand methaemoglobin levels up to 70% of
normal haemoglobin without significant mortalities (Smith
& Williams 1974; Brown & Mc Leay 1975). Moreover, cer-
tain fish species have acclimatization mechanisms with
acute exposure resulting in increased formation of methae-
moglobin and then normalize to basal levels upon long-
term exposure (Hilmy et al. 1987; Roques et al. 2013). Sev-
eral studies indicated that transferring a nitrite-affected fish
to clean water helps to restore the normal physiological
value of haemoglobin in 24–72 h (Huey et al. 1980; Knud-
sen & Jensen 1997). The effect of nitrite on the haemo-
lymph of crustaceans is variable and contradictory. Several
authors suggested that nitrite oxidizes haemocyanin in the
haemolymph to methaemocyanin and reduces the oxygen
transport capacity in Penaeus monodon (Cheng & Chen
2002), Macrobrachium rosenbergii (Chen & Lee 1997) and
Marsupenaeus japonicus juveniles (Cheng & Chen 1998).
However, the oxygen-carrying capacity was relatively unaf-
fected in other crustaceans, namely, freshwater crayfish
Astacus astacus (Jensen 1990b; Mallasen & Valenti 2006).
3
A. Ciji and M. S. Akhtar
Drastic decline in haemoglobin, haematocrit and ery-
throcyte count were commonly observed in nitrite-exposed
fish (Knudsen & Jensen 1997; Jensen 2003; Das et al. 2004a;
Ciji et al. 2012; Kim et al. 2018). Reduction in haemoglobin
content could be attributed to methaemoglobinaemia
(Knudsen & Jensen 1997; Jensen 2003) caused by nitrite as
well as depression and exhaustion of the haemopoiesis
under the hypoxic condition as suggested by Gill et al.
(1991). Another possible explanation for the decrease in
haemoglobin and haematocrit during nitrite intoxication is
the lysis of blood cells as suggested by Jensen (1990b) and
Knudsen and Jensen (1997). The formation of methaemo-
globin along with the reduced haemoglobin concentration
might have resulted in hypoxic condition within the fish.
Reduced haemoglobin and haemopoietic activity leading to
anoxia was reported in nitrite-exposed mrigal (Das et al.
2004a). Scarano and Saroglia (1984) suggested that the
increased energetic demand of RBCs due to higher activity
of NADH-methaemoglobin reductase to reconvert the
methaemoglobin to functional haemoglobin might have
resulted in a faster destruction of RBCs by the spleen and
kidney. The shrinkage of RBCs due to loss of potassium
ions to maintain K+ homoeostasis and the increased
removal of RBCs from the circulation (Jensen 1990b;
Knudsen & Jensen 1997) can be attributed to the reduction
in RBCs observed during nitrite exposure. Ciji et al. (2012)
observed a negative correlation between serum K+ and
haematocrit value in nitrite-exposed Labeo rohita.
Structural/morphological alterations
Fish gill is a multifunctional organ involved in excretion,
osmoregulation and respiration (Maetz 1971). The largest
surface area of gills is in direct contact with water making
them vulnerable to waterborne contaminants (Alvarado
et al. 2006; Romano & Zeng 2009; Luzio et al. 2013). It has
been reported that nitrite exposure induces clubbing, par-
tial/total fusion of secondary lamellae, hypertrophy and
hyperplasia of mucus and chloride cells, epithelial cell
necrosis, lifting of the epithelium and oedema leading to
heavy degeneration of gills (Deane & Woo 2007; Romano
& Zeng 2009; Wuertz et al. 2013). These morphological
alterations are linked to the physiology of the fish and ulti-
mately result in osmoregulatory dysfunction of gills (Deane
& Woo 2007; Park et al. 2007; Romano & Zeng 2009; Saoud
et al. 2014). However, nitrite exposure did not induce any
histopathological changes in the gills of silver perch, Bidya-
nus bidyanus (Frances et al. 1998).
The liver is a primary organ involved in the detoxifica-
tion of pollutants and exposure to different pollutants may
lead to structural damage (Pathan et al. 2010). Intracellular
oedema, cell necrosis or apoptosis, increases in sinusoidal
space, pycnotic nuclei, disintegration of hepatocytes,
4 © 2019 Wiley Publishing Asia Pty Ltd
formation of fat droplets and vacuolization were the major
histological changes seen in the livers of nitrite exposed
Megalobrama amblycephala and Cirrhinus mrigala (Than-
gam 2014; Sun et al. 2014b). In rainbow trout (Oncor-
hynchus mykiss), severe necrosis of the retina was observed
during nitrite exposure (Hofer & Gatumu 1994; Aggergaard
& Jensen 2001). Mild to moderate hyperplasia and hyper-
trophy of thyroid gland were evidenced in nitrate-exposed
white-spotted bamboo sharks, Chiloscyllium plagiosum
(Morris et al. 2011). Besides, Edwards and Guillette (2007)
suggested an increase in Leydig cell and testicular size/
weight during nitrate exposure. An increase in kidney
weight was reported in nitrite-exposed Sparus sarba and
Labeo rohita (Deane & Woo 2007; Ciji et al. 2013a). In tele-
osts, the kidney, along with the gills and intestine, plays a
significant role in the excretion and maintenance of
homoeostasis of the body fluids (Hinton et al. 1992; Evans
1993). Furthermore, teleost kidney acts as an excretory
route for the metabolites of a variety of xenobiotics (Than-
gam 2014). Acute and sublethal nitrite exposure resulted in
glomerular expansions, swelling of tubules, dilation of
glomerular capillaries and hyaline droplet degeneration in
the kidney of Cirrhinus mrigala (Thangam 2014). Park
et al. (2007) observed abnormal gross pathology in skin,
gill, liver and kidney tissues of dark-banded rockfish,
Sebastes inermis exposed to higher nitrite. The authors
reported necrosis of dermis and severe cytological destruc-
tion of hepatocytes and kidney cells.
Since crustaceans possess a hard and calcified exoskele-
ton that is quite impermeable to ions, gill is perhaps at high
risk to the toxic effect of environmental pollutants
(Pequeux 1995; Romano & Zeng 2013). Limited studies
have been undertaken relating the effect of nitrite on histo-
morphological alterations of gill in crustaceans. Romano
and Zeng (2009) reported disrupted pillar cells, increased
epithelial lifting and swelling as well as increased number of
haemocytes in the gill lamellae of blue swimmer crab, Por-
tunus pelagicus exposed to nitrite. The hepatopancreas, the
most important digestive gland in crustaceans, was found
to be remarkably disorganized during nitrite exposure. Han
et al. (2017) reported disorganized tubules with injured
connective tissues, blurred boundaries, decreased epithelial
cells, infiltrated haemocytes, narrowed lumen and vac-
uolization in the hepatopancreas of L. vannamei exposed
to higher ammonia/nitrite.
Disruption of ionic regulation
The alteration of ionic homoeostasis is well evidenced in
nitrite-exposed fish and crustaceans (Williams & Eddy
1986; Jensen et al. 1987; Harris & Coley 1991; Jensen 1993,
1996a; Stormer et al. 1996; Knudsen & Jensen 1997; Cheng
& Chen 1998). As nitrite is supposed to enter the fish and
Reviews in Aquaculture, 1–31

crustacean body by competing with chloride for the active
branchial chloride uptake through Cl
/HCO3
exchanger
(Williams & Eddy 1986), it can lead to disturbances in
extracellular electrolyte composition, principally with a
reduction in plasma chloride concentration (Jensen et al.
1987; Jenson 2003). Furthermore, accumulation of NO2-
disturbs K+ homoeostasis leading to loss of potassium from
skeletal muscle (Jensen 1990b; Stormer et al. 1996; Knudsen
& Jensen 1997) and red blood corpuscles (Jensen 1990a),
leading to extracellular hyperkalaemia in fish (Jensen et al.
1987). Aggergaard and Jensen (2001) have reported that
rainbow trout exposed to nitrite showed a rise in plasma
K+ and a decrease in plasma Cl-. In nitrite-exposed carps, a
significant reduction in plasma Na+ was observed (Jensen
et al. 1987; Ciji et al. 2012). Jensen et al. (1987) explained
that the significant reduction in plasma Na+ in nitrite
exposed carp might be due to the inhibition of Na+-K+-
ATPase activity. Furthermore, Tipsmark and Madsen
(2003) suggested inhibitory effect of nitric oxide on gill and
kidney Na+/K+-ATPase activity. Nitrite also inhibits car-
bonic anhydrase in trout gills (Gaino et al. 1984), and this
inhibition reduces the production of H+, the counter ion
for Na+ uptake, reducing the Na+ influx rate (Harris &
Coley 1991). However, an increase in plasma sodium levels
has been reported in Lates calcarifer exposed to nitrite
(Woo & Chiu 1997). A drastic decline in total osmolality,
plasma sodium and chloride was observed in nitrite-ex-
posed Channa striata (Lefevre et al. 2012). In contrast, total
osmolality remained unchanged in several species during
nitrite exposure (Jensen et al. 1987; Aggergaard & Jensen
2001; Jensen 2007; Lefevre et al. 2011; Ciji et al. 2013a).
According to Jensen et al. (1987), some adjustments in elec-
trolyte status occur during nitrite exposure to maintain
ionic homoeostasis or electroneutrality altered by the rise
in plasma NO2
and it is balanced either by a parallel rise
in cation or a parallel reduction in anion concentration.
Nitrite exposure resulted in reduced haemolymph Cl- and
Na+ levels in Pacifastacus leniusculus and Marsupenaeus
japonicas (Harris & Coley 1991; Cheng & Chen 1998). The
haemolymph osmolality was also found to be altered in
M. japonicas (Harris & Coley 1991), P. monodon (Chen &
Cheng 2000) and A. astacus (Jensen 1996b) during nitrite
exposure. The authors suggested that disruptions to Cl
/
HCO3
exchanger and increased water uptake might con-
tribute to haemolymph dilution and lowered Cl- and Na+
levels and osmolality.
Shift in metabolism/increased anaerobic substrate
oxidation
The formation of methaemoglobin along with reduced hae-
moglobin levels during nitrite intoxication compromises
the oxygen-carrying capacity of blood leading to hypoxic
Reviews in Aquaculture, 1–31
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Problems and management of nitrite in aquaculture
condition in tissues. Furthermore, nitrite is proven to
induce haemolytic anaemia (Scarano & Saroglia 1984),
which can aggravate the hypoxia stress (Moraes et al.
2006). Regulation of the glycolytic pathway has been sug-
gested as a metabolic strategy to endure hypoxia
(Hochachka & Somero 2002). Generally, during hypoxia, a
decrease in liver glycogen and an increase in liver lactate, a-
glycerophosphate and succinate was observed in teleosts
(Thillart et al. 1980; Jorgensen & Mustafa, 1980a,1980b;
Woo & Wu 1984; Yu & Woo 1987). Hybrid tambacu (Piar-
actus mesopotamicus 9 Colossoma macropomum), traira
(Hoplias malabaricus), common carp (Cyprinus carpio) and
oscar (Astronotus ocellatus) exposed to nitrite showed
increased lactate concentrations in the liver and plasma
indicating enhanced anaerobic oxidation of glucose under
hypoxia (Jensen 1992; Moraes et al. 1998; Moraes et al.
2006). Similarly, Arillo et al. (1984) reported a marked
decrease in blood glucose and liver glycogen and an
increase in liver and brain lactate of rainbow trout exposed
to nitrite. Furthermore, enhanced lactate dehydrogenase
(LDH) activity was observed in nitrite-exposed carps and
freshwater prawn (Das et al. 2004b; Jiang et al. 2013; Ciji
et al. 2014) indicating that pyruvate, the end product of
glycolysis, enters into anaerobic pathway leading to
increased lactate production. However, in Lates calcarifer,
significant reduction in liver glycogen was evident, but lac-
tate accumulation was noticed only when exposed to extre-
mely high nitrite concentrations (Woo & Chiu 1997).
Disruption of water balance
Despite several studies describing toxic effect of nitrite on
fish, its effect on water balance and water transport proteins
are not well addressed. Recent studies showed increased
reno-somatic index in Sparus sarba and Labeo rohita
exposed to higher nitrite (Deane & Woo 2007; Ciji et al.
2013a). Based on immunoassays, the authors suggested that
increased water retention in nitrite-exposed S. sarba is due
to reduced aquaporin 3 water channel proteins availability
in kidneys resulting in inefficient water extrusion (Deane &
Woo 2007). However, further detailed studies are required
to substantiate the effect of nitrite on aquaporin expression
and the resultant water retention. Similarly, Cheng and
Chen (1998) reported increased tissue water accumulation
in nitrite-exposed kuruma shrimp, Penaeus japonicas.
Oxidative damage and apoptosis
Nitrite exposure could enhance the intracellular formation
of reactive oxygen species (ROS) and reactive nitrogen spe-
cies (RNS) in aquatic animals and hence induces oxidative
damage (Jensen & Hansen 2011; Ciji et al. 2012; Guo et al.
2013; Romano & Zeng 2013; Sun et al. 2014a; Jensen et al.
5
A. Ciji and M. S. Akhtar
2015). Studies showed alteration of the antioxidant defense
system leading to the loss of antioxidant components com-
prising of superoxide dismutase (SOD), catalase, glu-
tathione peroxidase (GPx) and glutathione reductase
(GSH) during nitrite exposure (Apel & Hirt 2004; Bopp
et al. 2008; Jia et al. 2015). Upregulation of genes encoding
major antioxidative enzymes, namely, catalase, GPx, GSH,
glutathione-S-transferase and SOD, was noticed in nitrite-
exposed fish and shellfish as a coping strategy to increased
oxidative stress (Sun et al. 2014a; Zheng et al. 2016; Lin
et al. 2018a). The imbalance between antioxidant protec-
tion and free radical production was attributed to the toxi-
cological effect of nitrite in Macrobrachium rosenbergii
(Wang et al. 2004). Nitrite-induced oxidative stress
reported to cause impairment in membrane function and
loss of structural integrity due to oxidation of polyunsatu-
rated fatty acids (Wang et al. 2004). Higher malonaldehyde
(MDA) levels were also noticed in nitrite-exposed fish and
shrimp indicating prevalent oxidative stress and the resul-
tant depletion of antioxidant components (Uner € et al.
2001; Sun et al. 2014a; Jia et al. 2015; Han et al. 2017; Lin
et al. 2018a; Wang et al. 2018). Furthermore, the increased
ROS and NO formation upon nitrite exposure are also
known to initiate apoptosis (Jensen & Hansen 2011; Inad-
omi et al. 2012; Miao et al. 2018) through upregulation of
major transcription factor P53 and other apoptosis related
genes (Doman et al. 1999; Luzio et al. 2013; Guo et al.
2013; Cheng et al. 2015; Jia et al. 2015). Sun et al. (2014b)
reported nitrite-induced upregulation of apoptosis-related
genes, namely, caspase-8 and caspase-9 and enhanced
apoptosis in gills of Megalobrama amblycephala. Similarly,
previous studies demonstrated nitrite-induced haemocyte
apoptosis in P. monodon, L. vannamei and M. rosenbergii
via caspase-dependent pathway (Xian et al. 2011; Guo et al.
2013; Zhang et al. 2015). Furthermore, defender against
apoptotic death (DAD-1) is an anti-apoptotic gene
(Yamada et al. 2004) involved in the inhibition of
unwanted apoptosis (Lee et al. 2003; Zhu et al. 2008), and
its expression was found to be significantly affected by
nitrite exposure in M. japonicus (Zheng et al. 2016).
Reduction of immunocompetence
Nitrite exposure is known to suppress the immune
responses of fish and shellfish and makes them susceptible
to various infections (Cheng et al. 2002; Tseng & Chen
2004; Ciji et al. 2013b, 2015). The major defence compo-
nents such as lysozyme, complements and immunoglobulin
(IgM) play critical role in protecting fish against pathogens
and harmful conditions (Tort et al. 2003). Earlier investiga-
tions reported that nitrite exposure caused reduction in
lysozyme activity and IgM levels in fish compromising the
immune system (Ciji et al. 2015; Jia et al. 2016).
6 © 2019 Wiley Publishing Asia Pty Ltd
Furthermore, activation of complement system was evi-
denced in nitrite-exposed fish suggesting reduced immuno-
competence (Jia et al. 2016) as persistent activation of
complement could lead to immunosuppression (Peakman
et al. 1989; Yin et al. 2014). Significant reduction in total
leucocyte count observed in nitrite-exposed carps indicat-
ing failure or exhaustion of leucopoiesis further substanti-
ating immunosuppressive effect of nitrite (Das et al. 2004a;
Ciji et al. 2013a). Another potential indicator of stress, the
heat shock proteins (HSPs) which function as molecular
chaperones, are involved in folding and unfolding of newly
synthesized and of denatured proteins, respectively (Iwama
et al. 1999), are known to participate in the immune
response (Viant et al. 2003; Kim et al. 2013; Zhang et al.
2015). Hence, increased levels of HSPs (HSP70 and HSP90)
may reflect protein damage or increased tolerance to subse-
quent stress (Iwama et al. 1998). Several researchers
demonstrated enhanced expression of HSP70 and HSP90
in fish exposed to nitrite (Deane & Woo 2007; Sun et al.
2014b; Jia et al. 2016). Toll-like receptor-3 (TLR-3) is one
of the innate immunity-related pattern recognition recep-
tors and the activation of which induces the expression and
secretion of various cytokines to regulate inflammatory
response (Lee et al. 2014). Jia et al. (2016) reported
increased expression of cytokine genes (IL-1b and TNF-1a)
in the gills of the nitrite-exposed turbot indicating nitrite-
induced inflammatory response.
Production and release of haemocytes into the haemo-
lymph is one of the most essential immunological
responses of crustaceans as they possess only innate immu-
nity (Johansson et al. 2000; Ellis et al. 2011). Reduced total
haemocyte count, phagocytosis and phenoloxidase activity
were reported in a number of crustaceans exposed to higher
levels of nitrite indicating different magnitude of immuno-
suppression (Cheng et al., 2002; Tseng & Chen 2004;
Chand & Sahoo 2006; Xian et al. 2011; Romano & Zeng
2013). Furthermore, Zheng et al. (2016) reported a down-
regulation of immune genes, namely, lysozyme and stylicin
in Marsupeneaus japonicus indicating a weakened immune
response during nitrite exposure. Freshwater prawn Macro-
brachium rosenbergii and Macrobrachium malcolmsonii were
found to be more susceptible to nitrite toxicity than marine
shrimp (Wickins 1976; Chand & Sahoo 2006). Apoptosis of
immune-related tissues was enhanced at higher nitrite lead-
ing to immunosuppression in P. monodon and L. vannamei
(Xian et al., 2002; Tseng & Chen 2004; Xian et al. 2011;
Guo et al. 2013).
The nutrient dense water in the intensive culture systems
and RAS harbours a wide diversity of microorganisms. It
has been evident that the accumulation of nitrogenous
compounds has negative effects on ammonia and nitrite
oxidizers as Auffret et al. (2013) noticed reduced abun-
dance of dominant nitrite oxidizer Nitrospira in the RAS at
Reviews in Aquaculture, 1–31
 Problems and management of nitrite in aquaculture

higher levels of nitrite. The microbial compositions of rear-

ing water and diet greatly influences the shaping and colo-
nization of microbes in fish gut. Generally, Proteobacteria,
Bacteroidetes and Planctomycetes are normally dominant
in the intestines of shrimp and fish (Roeselers et al., 2011;
Rungrassamee et al., 2014; Xiong et al., 2015; Duan et al.,
2017; Dehler et al., 2017). Although the effect of rearing
environment/culture systems on gut microbiota is well doc-
umented, the impact of water nitrite in particular has not
been studied extensively in teleosts. One study in Litope-
naeus vannamei by Duan et al. (2018) showed a decrease in
the intestinal bacterial diversity in response to ammonia
and nitrite exposure. The authors noticed a shift in the
dominant bacterial phyla from Proteobacteria to Bac-
teroidetes, and a simultaneous decrease in Planctomycetes
in ammonia and nitrite-exposed shrimp. Furthermore, a
decrease in the symbiotic Photobacterium and an increase
in the opportunistic pathogens including Nautella and
Pseudoalteromonas were also noticed. By altering the gut
microbial composition, the intestinal immunity is compro-
mised in ammonia and nitrite-exposed Litopenaeus van-
namei (Duan et al., 2018).
Endocrine disruption
The effect of nitrite on endocrine functions and reproduc-
tive health of fish is received considerable attention only in
recent years. In vivo and in vitro studies in mouse already
proved that intake of nitrite and nitrate causes steroid hor-
mone deficiency (Panesar & Chan 2000). The anticipated
mechanisms (Fig. 1) for nitrite/nitrate induced reproduc-
tive dysfunctions include mitochondrial conversion to
nitric oxide (NO) and the resultant alterations in enzymatic
actions (Guillette & Edwards 2005; Edwards & Hamlin
2018). Low pH and tissue hypoxia due to increased lactate
and methaemoglobin levels proved to result in the forma-
tion of nitric oxide from nitrite (Zweier et al. 1999) which
can potentially inhibit several components of the steroido-
genic pathway (Fig. 1), including steroidogenic acute regu-
latory protein (StAR), cytochrome P450-sidechain cleavage
enzyme (SCC) and 3b-hydroxysteroid dehydrogenase
(3bHSD) (Delaforge et al. 1995; DelPunta et al. 1996; Pane-
sar & Chan 2000). It was suggested that the NO reacts with
iron complexes of cytochrome p450 series of enzymes pre-
sent at multiple locations along the steroidogenic pathway

Figure 1 Schematic diagram presenting probable mechanism of endocrine (reproductive) disruption by nitrite in fish [StAR: Steroidogenic acute reg-
ulatory protein; P450cc: Cytochrome P450-sidechain cleavage enzyme; 3/ 17 bHSD: 3/ 17b-hydroxysteroid dehydrogenase; P450 17a: Cytochrome
P450 17a-hydroxylase; P450 arom: Cytochrome P450 aromatase; Hb: Haemoglobin; MetHb: Methaemoglobin; -Ve: Inhibitory action of nitric oxide;
+Ve: Stimulatory action of nitric oxide; ??: Possibility of either inhibitory or stimulatory effect by NO that needs to be investigated].

Reviews in Aquaculture, 1–31

© 2019 Wiley Publishing Asia Pty Ltd 7
A. Ciji and M. S. Akhtar
(DelPunta et al. 1996; Weitzberg & Lundberg 1998; Panesar
& Chan 2000). However, its inhibitory role on other
enzymes of the steroidogenic pathway such as p450 17a-hy-
droxylase (p450 17a) and 17b-hydroxysteroid dehydroge-
nase (17bHSD) is still needs to be investigated. As these
two enzymes share common properties with cyctochrome
p450 series and steroid dehydrogenase enzymes, there are
fair chances that NO has an inhibitory role in their activi-
ties as well. Hypoxia itself acts as an endocrine disruptor in
fish and serum levels of testosterone, estradiol and tri-
iodothyronine significantly decreased in carp Cyprinus car-
pio exposed to chronic hypoxia (Wu et al. 2003). Freitag
et al. (2015) reported that moderate concentrations of envi-
ronmental nitrate (10.3 mg L
1) increased the synthesis of
testosterone in Atlantic salmon while elevated concentra-
tions (101.8 mg L
1) inhibited testosterone synthesis
through increased formation of NO in the gonadal tissues.
The authors speculated that by inhibiting aromatase,
another P450 enzyme, the conversion of testosterone to
estradiol might have reduced at moderate nitrate concen-
trations resulting in higher plasma testosterone levels.
Nitrite exposure was found to impair steroid hormone syn-
thesis (both testosterone and estradiol) in Labeo rohita (Ciji
et al. 2013c) juveniles (Fig. 2). Recently, Lin et al. (2018b)
reported significant reduction in testosterone levels in
nitrite-exposed zebrafish through downregulation of the
expression of GnRH and LH genes. Nitrate affects sper-
matogenesis either indirectly through alterations of
Figure 2 Effect of nitrite on thyroid and reproductive hormones of
Labeo rohita (Ciji et al. 2013c). Mean values bearing different super-
scripts (a, b, c) in the series vary significantly (P < 0.05). Data expressed
as mean  SE, n = 6. , 0 mg NO2-N/L; , 2 mg NO2-N/L
8 © 2019 Wiley Publishing Asia Pty Ltd
steroidogenic enzyme or directly by regulating sperm cell
apoptosis (Edwards & Guillette 2007) as nitric oxide is
implicated in apoptosis of spermatogonia, spermatocytes
and spermatids (Zini et al. 1996; El-Gohary et al. 1999;
Lue et al. 2003; Di Meglio et al. 2004). Reduction in
gonadosomatic index and testis weight was noticed in
nitrite-exposed green sea urchin and zebrafish (Siikavuo-
pio et al. 2004; Lin et al. 2018b). A number of vertebrate
studies showed that exposure to nitrate/nitrite or other
NO donors reduces reproductive hormone concentra-
tions, fertilization and pregnancy rates, dry embryo
weights and semen quality, suggesting a direct negative
effect on reproductive performance (DelPunta et al. 1996;
Guillette & Edwards 2005; Edwards et al. 2006a; Edwards
et al. 2006b). Nevertheless, limited literature is available
relating the effect of nitrite on fish steroidogenesis and
reproduction. Edwards and Guillette (2007) observed an
increase in testicular weight and a reduction in sperm
count in nitrite-affected mosquito fish Gambusia hol-
brooki without a significant reduction in testosterone
indicating that the increase in the Leydig cell would
account for the increased testicular size and the conse-
quent maintenance of testosterone concentrations.
In addition to sex steroids, nitrite is known to reduce
the production of thyroid hormones in Sparus sarba
(Deane & Woo 2007) and L. rohita (Ciji et al. 2013c)
juveniles (Fig. 2). However, white- spotted bamboo sharks
(Chiloscyllium plagiosum) reared in 70 mg L
1 NO3-N for
29 days did not manifest a significant reduction in
plasma-free thyroxine (T4), but showed mild to moderate
hyperplasia and hypertrophy of the thyroid gland (Morris
et al. 2011). Similarly, plasma thyroid levels in Atlantic
salmon was unaffected by nitrate exposure (Freitag et al.
2015). Mukhi et al. (2005) explained that T4 could be
stored within the colloid of thyroid follicles, and there-
fore, a reduction in plasma T4 can only be observed after
the exhaustion of stored T4 in the follicles. Low levels of
nitrate in drinking water resulted in a significant reduc-
tion in thyroid hormone levels in Wistar rats (Eskiocak
et al. 2005). Earlier studies described nitrate as a goitrogen
which can competitively bind to the sodium/iodide sym-
porter on thyroid follicles leading to reduced availability
of iodine for thyroid hormone synthesis (Lahti et al. 1985;
Tonacchera et al. 2004; De Groef et al. 2006). Thyroid
hormones play vital roles in growth, development, repro-
duction and stress responses in fish (Higgs et al. 1977;
Leatherland 1994; Power et al. 2001; Peter 2007). Ciji
et al. (2013c) reported a positive correlation between
growth rate and serum thyroid hormone levels in L. ro-
hita juveniles. Although the effect of nitrite exposure on
fish growth is extensively studied its adverse effect on thy-
roid hormones, the key metabolic regulator is not yet
comprehensively studied.
Reviews in Aquaculture, 1–31

Flesh quality
Fish raised in recirculatory aquaculture systems (RAS) can
develop ‘earthy’ and/or ‘musty’ off-flavours due to the
accumulation of geosmin and 2-methylisoborneol, respec-
tively, which can adversely affect the quality and market
value of the fish (Tucker, 2000; Howgate, 2004). These off-
flavours are commonly observed in aquatic animals reared
in freshwater culture systems with high nutrient loads
(Tucker, 2000). Blevins et al. (1995) indicated increasing
geosmin/biomass production by Streptomyces halstedii with
increasing NO3-N concentrations contrary to Schrader
et al. (2013) who did not observe any difference in geosmin
concentrations in RAS waters or in rainbow trout flesh with
increasing NO3-N concentrations. Hence, further studies
are needed to uncover the exact source of off-flavours in
fish raised in RAS and to rule out the effect of nitrogenous
compounds on off-flavours. Guttman and van Rijn (2008)
found an increase in growth of actinomycetes and produc-
tion of off-flavour compounds under aerobic and organic-
rich conditions. A higher phosphate level (1 mg L
1) is
known to stimulate geosmin production in aquaculture
systems (Robertson et al., 2006; Auffret et al., 2013).
Mechanism of nitrite detoxification
Several mechanisms are proposed for detoxification of
nitrite. Fish erythrocytes contain NADH-methaemoglobin
reductase enzyme capable of reconverting methaemoglobin
to haemoglobin (Huey & Beitinger 1982). Hence, transfer-
ring a nitrite-poisoned fish to clean water helps in the grad-
ual restoration of normal proportion of haemoglobin
unless nitrite toxicity has progressed too far (Knudsen &
Jensen 1997; Kroupova et al. 2018). The elevated metHb
levels during prolonged nitrite exposure assumed to
increase the activity of methaemoglobin reductase resulting
in a reduction in nitrite concentrations with a concomitant
increase in plasma nitrate (Schoore et al. 1995; Knudsen &
Jensen 1997). Jensen and Nielsen (2018) suggested that the
higher metHb reductase activity in fish is an indication of
higher Hb autoxidation. Enhanced activity of methaemo-
globin reductase has been observed in nitrite-exposed B.
cephalus and L. rohita (Fig. 3) as a protective mechanism
by reducing Fe3+ in the haemoglobin to Fe2+ (Avilez et al.
2004; Ciji et al. 2012). Usually, nitrite is eliminated through
both gills and urine (Jensen 2003). Another detoxifying
mechanism of nitrite in fish is its oxidation to low toxic
nitrate (Doblander & Lackner 1997). Hence, nitrate con-
centrations in the body found to increase during nitrite
exposure unless excretion is concurrently increased (Dob-
lander & Lackner 1996, 1997; Stormer et al. 1996). The
enzymes, particularly catalase and cytochrome oxidase, are
proved to be the major routes of detoxification of nitrite to
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Figure 3 Effect of nitrite concentration on methaemoglobin reductase
activity in blood of Labeo rohita (Ciji et al. 2012). Mean values bearing
different superscripts (a and b) in both the nitrite concentrations vary
significantly (P < 0.05). Data expressed as mean  SE, n = 6.
nitrate (Kroupova et al. 2005). Crustaceans also have the
ability to detoxify nitrite by converting it into nitrate (Chen
& Cheng 1995; Jensen 1996b; Cheng & Chen 1998).
Factors affecting nitrite toxicity
Nitrite toxicity varies considerably depending on a large
number of external and internal factors (Table 1). Toxicity
differs noticeably across different species of fish. Brackish
water and marine fish are less susceptible to nitrite toxic-
ity. Fish species with high branchial chloride uptake rate
(e.g. rainbow trout, perch, channel catfish and pike) are
more sensitive to nitrite than fish species with low chloride
uptake (e.g. carp, blue gill and eel) (Perrone & Meade
1977; Bath & Eddy 1980; Gutzmer & Tomasso 1985; Wil-
liams & Eddy 1986; Jensen 1995). Salmonids are the most
sensitive among the freshwater fish species studied (Krou-
pova et al. 2018). It also varies depending on the duration
of nitrite exposure, water quality as well as fish size and
age (Williams & Eddy 1988; Aggergaard & Jensen 2001;
Jensen 2003; Kroupova et al. 2018). Some fish species
show intraspecific variability in nitrite uptake and its sus-
ceptibility (Margiocco et al. 1983; Jensen 2003). These dif-
ferences between individuals were often observed in
rainbow trout (Margiocco et al. 1983; Jensen 2003), and it
may be attributed to differences in the number of chloride
cells and its exposed surface area or difference in the pro-
liferation of chloride cells (Kroupova et al. 2005). Agger-
gaard and Jensen (2001) suggested that 24- to 48-h
exposure is required for maximum accumulation of nitrite
in fish. Different water quality parameters such as chlo-
ride, other anions (bromides) and cations (calcium, mag-
nesium, sodium and potassium), ammonia, pH, oxygen
and temperature are known to affect the susceptibility of
fish to nitrite toxicity (Kroupova et al. 2005). The older
9
A. Ciji and M. S. Akhtar

Table 1 Effects of other water quality parameters on nitrite levels/toxicity

Parameter Effect Reference

Chloride (Cl-) • With increase in Cl concentration nitrite toxicity decreases.

-
Russo and Thurston
concentration • The relationship between nitrite toxicity and chloride concentration is reported to
be linear in many teleosts.
(1977); Palachek and
Tomasso (1984b);
• Calcium chloride is found to be more effective than sodium chloride at reducing
nitrite toxicity in fish.
McConnell (1985);
Kroupova et al. (2005)
Mazik et al. (1991)
Fontenot et al. (1999)
Bromide (Br-) • Decreases the toxicity of nitrite. Eddy et al. (1983)
• 10.7mMmMofnitrite
1
sodium bromide (80 mg L ) was enough to offset the presence of

1
(32 mg L nitrite-N) for Atlantic salmon (Salmo salar) in freshwa-
ter.
pH • The effect of pH on nitrite toxicity within the natural/optimum pH range of fish
seems to be very little.
Lewis and Morris
(1986); Wedemeyer
• The fraction of HNO in a given nitrite concentration increases with decrease in pH
2
level. Hence, the nitrite-induced toxicity increases as it is considered to be depen-
and Yasutake (1978)
Chen and Cheng
dent on the concentration of HNO2.
• Bath
(2000) Meade and
and Eddy (1980) reported that pH had no significant on nitrite toxicity in rain- Perrone (1980)
bow trout except at extreme pH levels (<5.0 and >10).
• Aanism
change in the pH can alter the uptake characteristics of ion-concentrating mech-
in the gills, leading to pH effects over very large pH ranges.
Temperature • lamellar water
Increase in temperatures enhances the nitrite uptake in fish by way of increased
flux and ionic exchange through the gill and thus increases the
Huey et al. (1984)
Lewis and Morris
nitrite toxicity. (1986)
• Over a small range of temperature (22–30°C), Colt and Tchobanoglous (1976)
found no significant relationship between temperature and nitrite toxicity.
• Channel
1
catfish kept at 30°C in the presence of 0.91 mg L nitrite-N over a 24-h
built-up methaemoglobin concentrations twice as high as those of fish held at
10°C.
• However, it is found that lower temperatures reduce the efficiency of detoxification
mechanisms of nitrite.
Dissolved oxygen • Oxygen may affect nitrite toxicity because nitrite reduces the oxygen-carrying
capacity of blood.
Bowser et al. (1983)

• Low dissolved oxygen can diminish the bacterial nitrifying process to convert
ammonia to nitrite.
• Infishthe presence of nitrite, oxygen demand was comparatively higher in channel cat-
Ammonia • No synergistic or additive effects on plasma corticosteroid levels were observed
when the channel catfish were exposed to ammonia and nitrite in combination.
Tomasso et al. (1981)
Vedel et al. (1998)
• Combined nitrite and ammonia exposure resulted in high mortality in rainbow trout
at the highest exposure concentrations (600 l nitrite x 500 l ammonia).
M M

• Enhances nitrite accumulation and triggers potassium loss in teleost.

Alkalinity • Nitrifying
tion process.
bacteria use bicarbonate as carbon source for their growth and nitrifica- Paz (1984) and
Biesterfeld et al.
• Maintaining
1
adequate alkalinity concentrations in the range of 40–80 mg L (as
CaCO ) has been reported to be critical for sustaining nitrification.
(2003) Chen et al.
3 (2006)
• Itfication
1
is recommend to maintain an alkalinity of 200 mg L as CaCO to support nitri-
when water exchange rate is minimal.
3

Divalent and trivalent • Have very low effects on nitrite toxicity. Lewis and Morris
anions (such as sulphate, (1986) Russo et al.
phosphate and borate) (1981)

fish are more susceptible to nitrite toxicity as compared to
smaller fish possibly because of higher activity of detoxify-
ing enzyme methaemoglobin reductase in smaller fish
(Kiese 1974; Kroupova et al. 2018). However, Siwicki et al.
(1994) suggested that a higher oxygen-binding capacity of
haemoglobin in young trout as compared with older ones
makes small ones less susceptible to nitrite stress. Further-
more, Kroupova et al. (2018) suggested that, in early
developmental stages, respiration occurs mostly through
skin thus lowering the importance of oxygen transport
through blood. Increased tolerance of younger fish to
higher nitrite concentrations has been demonstrated in

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10 © 2019 Wiley Publishing Asia Pty Ltd

several fish species Oncorhynchus kisutch (Perrone &
Meade 1977), Salmo gairdneri (Smith & Williams 1974;
Russo et al. 1974; Russo & Thurston 1977), Salmo trutta
(Bartlett & Neumann 1998), Pimephales promelas (Pala-
chek & Tomasso 1984a,1984b), Oreochromis niloticus
(Atwood et al. 1996) and Danio rerio (Voslarova et al.
2006).
Remedial/mitigation measures
Since nitrite toxicity is inevitable in aquaculture systems,
different approaches and management strategies (Fig. 4;
Tables 2 and 3) have been evolved and evaluated to address
the nitrite-associated stress responses. The major remedial/
mitigation measures are discussed below.
Salt addition
Adding salt preferably chloride is the most common and
standard practice for reducing the adverse effect of nitrite
toxicity. Increasing salinity of the rearing water is shown to
be effective in mitigating the toxicity of ammonia and
nitrite (Table 1) to fish (Bianchini et al. 1996; Sampaio
et al. 2002; Roques et al. 2013) due to its competitive effects
on branchial nitrite uptake (Crawford & Allen 1977). In
Figure 4 Strategies to remove nitrite or mitigate its toxicity in aquaculture systems.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
freshwater, nitrite is more toxic (~55 times) to milkfish
Chanos chanos in comparison to those reared in 16 ppt
brackish water (Almendras 1987). The 96-h LC50 of nitrite
to seabass (Lates calcarifer) was 14.5 mg L
1 in 0 ppt while
it was recorded as 105 mg L
1 in 15 ppt (Woo & Chiu
1994). Bath and Eddy (1980) postulated that the gill chlo-
ride cells in freshwater teleosts have a low affinity for nitrite
ions, and hence, nitrite uptake can be competitively inhib-
ited by external chloride ions. Pistekova et al. (2005)
observed a logarithmic relationship between nitrite toxicity
and chloride concentrations. Tucker and Hargreaves (2004)
suggested that the application of chloride in the form of
common salt at a ratio of chloride to nitrite of 30:1 pre-
vents nitrite toxicity and associated physiological alter-
ations. However, this protective effect is not uniform
between species (reviewed by Lewis & Morris 1986). The
European Inland Fisheries Advisory Commission (EIFAC)
recommends a weight ratio of chloride to nitrite-nitrogen
for salmonids/rainbow trout and cyprinids as 17:1 and 8:1
respectively. However, later case studies by Svobodova et al.
(2005b) showed incidence of health impairment and mor-
talities in different fish species (catfish, tench and tilapia) at
chloride to nitrite-nitrogen ratio well above the optimum
levels of EIFAC (1984). The recommended ratio of chloride
to nitrite-nitrogen for different commercial fish species is
11
A. Ciji and M. S. Akhtar

Table 2 Different management/mitigation approaches adopted for reducing nitrite toxicity in fish and shellfish

Management approach Fish/shellfish species NO2- N (mg/L) Responses Reference

Salt/chloride addition
20 ppt salinity Takifugu obscurus 20 mg L
1 NO2
–N for 96 h Higher survival, reduced Wang et al. (2018)
(Puffer fish) methaemoglobin and MDA
formation, increased SOD,
catalase and GSH activities,
increased expression of
antioxidant genes, increased gill
Na+K+ATPase activity and tissue
HSP70
≥240 mg L
1 chloride Sander lucioperca (Pike 10 mg L
1 NO2
–N for 42 days Inhibited nitrite accumulation in Wuertz et al. (2013)
as NaCl perch) plasma and muscle; reduced
methaemoglobin formation
10.1 mg L
1 chloride Orconectes limosus 40 mg L
1 NO2
–N for 96 h Increased tolerance; higher Kozak et al. (2005)
as NaCl (Spiny-cheek crayfish) survival
261.3 mg L
1 chloride Oncorhynchus kisutch 29.8 mg L
1 NO2
–N for 48 h Ameliorated stress; lowered Perrone and Meade
(Cl-) (Coho salmon) mortality and methaemoglobin (1977)
levels
Sodium chloride/ Oreochromis niloticus 10 mg nitrite-N per L for 96 h Increased the LC50; higher Atwood (2001)
calcium chloride was (Nile tilapia) survival and lowered plasma
added to raise the nitrite concentrations
chloride concentration
by 10 mg L
1
Sodium chloride was Clarias batrachus 78.7 mg L
1 NO2
–N for 48 h Increased the LC50; higher Duangsawasdi and
added at 250 mg L
1 survival Sripoomun (1981)
70 mg L
1 chloride Oreochromis niloticus 100 mg nitrite-N per L for 96 h An increase in the LC50 and Wang et al. (2006b)
(Nile tilapia) reduced mortality
100 mg L
1 chloride Danio rerio (Zebrafish) ~680 mg nitrite-N per L for 96 h Increased the LC50, higher Voslarova et al.
survival (2006)
50 mg Cl- per L Macrobrachium 10 mg nitrite-N per L for 24 h Increased oxyhaemocyanin, Chen and Lee
rosenbergii (Giant haemolymph protein, osmolality, (1997)
freshwater prawn/ chloride, Na+, K+ & Ca+ and
Scampi) decreased nitrite concentration
Nutritional manipulations/interventions
Dietary Labeo rohita (Rohu) 2.0 mg L
1 Augmented growth; increased Ciji et al.
supplementation of haemoglobin and haematocrit; (2013a, 2013b,
300 mg kg
1 reduced oxidative damage; 2013c, 2014)
a-tocopherol acetate reduced nitrite accumulation in
for 45 days muscle; increased activity of
methaemoglobin reductase;
improved immunity and disease
resistance
Dietary Labeo rohita (Rohu) 2.0 mg L
1 Augmented growth, reduced Ciji et al. (2013c,
supplementation of oxidative damage; improved 2015)
1.5% L-tryptophan for immunity and disease resistance
45 days
Dietary administration Macrobrachium 21.0 mg L
1 sodium nitrite Increased survival; enhanced Chand et al. (2006)
of 300 mg kg
1 rosenbergii (Giant innate immune responses by
bovine lactoferrin for freshwater prawn/ increasing phenoloxidase
7 days Scampi) activity, total protein and
agglutinin levels
Modification of pond bottom
Application of humic Danio rerio (Zebrafish) 21. 1 mMol nitrite-N for 144 h Higher survival Meinelt et al. (2010)
substances

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12 © 2019 Wiley Publishing Asia Pty Ltd
 Problems and management of nitrite in aquaculture

Table 3 Microbial mitigation/biological approaches for removal of toxic nitrite

Macro/microorganisms Cultured species Removal efficiency References

Bacteria – Vibrio alginolyticus
Bacteria – Klebsiella oxytoca
Bacteria – Nitrobacter winogradskyi
Bacteria – Nitrobacter
Bacteria – Nitrospira marina and
N. moscoviensis
Cyanobacterium – Oscillatoria
Mixed diatoms – Amphora, Cymbella
and Navicula
Seaweed – Ulva pertusa
Macroalgae – Gracilaria sp.
Shrimp
– Reduced NO2-N concentration from 10 mg L
1
Litopenaeus vannamei
Penaeus monodon
Litopenaeus vannamei
Penaeus monodon hatchery
Penaeus monodon hatchery
Sea cucumber (Apostichopus
japonicus)
Penaeus monodon
Degraded NO2-N from 11.41 to 5.46 mg L
1 Shan et al. (2016)
over 12 h in shrimp culture ponds
Liu et al. (2013)
to zero over 20 h in simulated condition
Mean NO2-N remained below 1.0 mg L
1 in the Kuhn et al. (2010)
Nitrobacter-treated systems and steadily
increased and reached 30 mg L
1 on day 20 in
untreated systems
NO2
concentration of the treatments with Hao et al. (2017)
Nitrobacter inoculation decreased from
0.06 mg L
1 on the first day to 0.03 mg L
1 on
the 16th day while NO2
concentration of the
treatments without Nitrobacter inoculation
increased from 0.06 mg L
1 on the initial day
to 0.13 mg L
1 on 16th day
– Brown et al. (2013)
0.07 g m
2 day
1 Khatoon et al. (2007)
0.06 g m
2 day
1 Khatoon et al. (2007)
0.45 g N m
2 day
1 Wang et al. (2007)
Promoted nitrite removal after 2–3 days of the Hao et al. (2017)
experiment

listed in Table 4. Russo and Thurston (1977) reported that Table 4 The recommended minimum weight ratio of chloride to
increase in chloride concentration by 1 mg L
1 resulted in nitrite-nitrogen for different commercial fish species
an increase in 96-h LC50 value by 0.29 mg L
1 N-NO2 in
Fish species Chloride to Reference
rainbow trout. However, salt addition/increasing salinity is
NO2-N ratio
practically feasible for fish that can tolerate high salinity
and chloride ions. In freshwater fish, increasing salinity Coho salmon (Oncorhynchus 17:1 Perrone and
itself induces stress due to osmotic pressure variations and kisutch) yearling Meade (1977)
can lead to physiological imbalances. The rate of formation Coho salmon (Oncorhynchus 4:1 Perrone and
kisutch) fry Meade (1977)
of methaemoglobin is directly proportional to the plasma
Channel catfish (Ictalurus 20:1 Tomasso et al. (1980)
nitrite concentration which in turn depends on water chlo- punctatus)
ride content in several fish species (Schwedler & Tucker Common carp 5:1 Hasan and
1983; Williams & Eddy 1988; Cheng & Chen 1999). The (Cyprinus carpio) fry Macintosh (1986)
effect of waterborne chloride on plasma nitrite and methae- Tilapia (Oreochromis niloticus) ≥150:1 Svobodova
moglobin levels in different fish species are depicted in Fig- et al. (2005a)
ures 5 and 6 respectively. Roques et al. (2013) observed a Milk fish (Chanos chanos) 15:1 Almendras (2014)
Pike perch (Sander ≥24:1 Wuertz et al. (2013)
potential attenuating effect of the addition of sodium chlo-
lucioperca)
ride on the feed intake of African catfish, Clarias gariepinus. Salmonids (rainbow trout, brook ≥100:1 Buric et al. (2016)
Wang et al. (2018) observed an explicit interaction between trout, brown trout, Siberian
nitrite and salinity on the responses of Hb, MetHb, antioxi- sturgeon, Russian sturgeon)
dant defence, Na+/K+ATPase, HSP70 and cortisol indicat-
ing a reduction in the impact of nitrite with increasing
salinity. The authors also hypothesized that although salin- (Sowers et al. 2004; Romano & Zeng 2013; Ramiırez-
ity may reduce the decline of RBCs caused by nitrite, RBCs Rochin et al. 2017).
abundance is also reduced by osmotic pressure caused by Chloride levels can be increased by adding commercially
increased salinity in the estuarine puffer fish Takifugu available sodium chloride (common salt) or calcium chlo-
obscurus. Several studies in crustaceans indicated that at ride. The effectiveness of chloride source in extenuating
lower salinities, the toxicological effect of nitrite increases nitrite toxicity varies between species. For instance, in

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© 2019 Wiley Publishing Asia Pty Ltd 13
A. Ciji and M. S. Akhtar

Figure 5 Effect of waterborne chloride (Cl
) on plasma nitrite-nitrogen levels in different teleost species: The trends of plasma nitrite-N reduction
were as in (i) Coho salmon Y =
4.78x + 57.1, R2 = 1; Meade and Perrone (1991), (ii) Shortnose sturgeon Y =
16.988x + 116.36, R2 = 0.834;
Fontenot et al. (1999), (iii) Pike perch Y =
0.0887x5 + 6.0061x4
160.71x3 + 2122.8x2
13845x + 35695, R2 = 0.966; Wuertz et al. (1997),
(iv) Rainbow trout Y =
18.637x + 299.3, R2 = 0.991; Eddy et al. (1983) (units were converted) and (v) Stripped bass Y =
5.6429x + 153.07,
R2 = 0.9995; Mazik et al. (2012). , Shortnose strugeon (2.23 mg/L NO2-N); , Rainbow trout (42 mg/L NO2-N); , Stripped bass (250 mg/L
NO2-N); , Pike perch (10 mg/L NO2-N); , Coho salmon (10 mg/L NO2-N)

Figure 6 Effect of waterborne chloride (Cl
) on nitrite-induced methaemoglobin levels in different teleost species: The trends of metHb reduction
were linear as in (i) Coho salmon – Y =
7x + 68.4, R2 = 1; Meade and Perrone (1991), (ii) Atlantic sturgeon – Y =
7.75x + 83.5, R2 = 1; Matsche
et al. (2012), (iii) Channel catfish – Y =
7.43x + 93.377, R2 = 0.969; Bowser et al. (1983), (iv) Tilapia – Y =
6.6x + 118.2, R2 = 1; Yanbo et al.
(2002), (v) Common carp – Y =
5.3x + 112.2, R2 = 1; Svobodova et al. (2009), (vi) Pike perch – Y =
7.2006x + 146.25, R2 = 0.9414; Wuertz
et al. 1997 and (vii) Stripped bass – Y =
6.2143x + 171.57, R2 = 0.9902; Mazik et al. 2012. , Channel catfish (5 mg/L NO2-N); , Tilapia
(15 mg/L NO2-N); , Stripped bass (250 mg/L NO2-N); , Pike perch (10 mg/L NO2-N); , Common carp (67 mg/L NO2-N); , Coho salmon
(10 mg/L NO2-N); , Atlantic sturgeon (1 mg/L NO2-N)

certain fish such as steelhead trout (Salmo gairdneri; Wede- brevirostrum; Fontenot et al. 1999), striped bass (Morone
meyer & Yasutake 1978) African sharptooth catfish, Clarias saxatilis; Mazik et al. 1991) and sunshine bass (hybrid of
lazera; Hilmy et al. 1987), shortnose sturgeon (Acipenser female white bass Morone chrysops X male striped bass

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14 © 2019 Wiley Publishing Asia Pty Ltd

Morone saxatilis; Weirich et al. 1993) calcium chloride was
reported to be more effective than sodium chloride. How-
ever, in tilapia (Atwood et al. 1996) and channel catfish
(Tomasso et al. 1980), both sodium chloride and calcium
chloride appeared to be equally effective in inhibiting
nitrite uptake. A twofold rise in 96-h LC50 of nitrite was
noticed in steelhead trout by addition of 65 mg L
1 chlo-
ride as sodium chloride while this increase was 35-fold
when the same concentration of chloride was maintained
by using calcium chloride (Wedemeyer & Yasutake 1978).
This difference had potentially been attributed to the ability
of calcium to alter the membrane permeability leading to
reduced nitrite diffusion, another route of nitrite uptake in
certain fish species (Potts & Fleming 1970; Fleming 1974).
Water exchange
One of the common practices for nitrite removal is regular
water exchange (Losordo & Timmons 1994; Boyd & Tucker
1998). However, water exchange in large quantum is labori-
ous and expensive leading to increased cost of production
in aquaculture. It also increases the risk of widespread of
diseases and eutrophication in rivers and coastal waters.
Therefore, the release of effluent water containing high
levels of toxic nitrite to the environment is non-sustainable
(Van Breemen & Van Dijk 1988) and cause biosafety issues.
Hence, water exchange is not advisable to address nitrite
toxicity in aquaculture unless cultured species is in critical
conditions. However, it is one of the most recommended
practices to avoid nitrite toxicity in aquarium or ornamen-
tal fish rearing units.
Electrochemical method for nitrite removal
The major consequence of aquaculture intensification is
the generation of aquaculture wastewater which is princi-
pally characterized by high loads of nitrogenous com-
pounds including ammonia, nitrite and nitrate (Petrilli &
Tonukari, 1997). Nitrite results in the wastewater effluents
of aquaculture systems due to fish excretion and decompo-
sition of uneaten feed (Poxton & Allhouse, 1982). Nitrite
can also react with other compounds such as amines and
amides leading to the production of cancer-causing com-
pounds like nitrosamines (Liao & Mayo, 1974). Further-
more, nitrite in wastewater results in the emission of a
green house gas nitrous oxide (N2O) to the environment
contributing to global warming (Foley et al., 2010). Treat-
ment of the wastewater including aquaculture wastewater
has become a grave issue for the environmentalists
throughout the world (Boggs and Botte, 2009; Saleem &
Essa, 2010). Furthermore, effluents from the aquaculture
industry are currently battling with stricter legal regulations
(Diaz et al., 2010) as the presence of high concentrations of
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Problems and management of nitrite in aquaculture
these toxic nitrogenous compounds causes a plethora of
troubles in the receiving aquatic environment. Therefore,
cost-effective and environment-friendly treatment of aqua-
culture wastewater is need of the hour for sustainable aqua-
culture development.
The electrochemical method of wastewater treatment has
been successfully employed to remove various pollutants
from industrial wastewaters (Lin & Wu, 1996b; Abuzaid
et al., 1999; Chen, 2004; Emamjomeh & Sivakumar, 2009).
Electrochemical technique can also be applied for nitrite
removal (Chakrabarti et al., 2011). Treatment of aquacul-
ture wastewater using electrochemical method has received
some attention recently (reviewed by Mook et al., 2012)
due to its advantage of high treatment efficiency, no sludge
production and relatively low investment costs (Mook
et al., 2012). However, its implementation to manage aqua-
culture wastewater in large scale is very meagre. Therefore,
the possibility of employing electrochemical methods for
nitrite removal from aquaculture wastewater, especially
from intensified shrimp/fish farms needs to be explored on
industrial basis for environment-friendly aquaculture
development. It warrants comprehensive research pro-
grammes to be taken to standardize the process and upscale
it to address the aquaculture waste, especially nitrite prob-
lem. Nevertheless, governments should also mandatorily
advocate/enforce electrochemical method for aquaculture
wastewater treatment before approving intensified aquacul-
ture units.
Microbial mitigation/biological approaches for removal of
toxic nitrite
The efficient way to remove nitrite is through biological
approaches (Table 3) as compared to physical and chemical
methods that lead to pollution and unfavourable residues
formation and accumulation (Van Rijn et al., 2006). How-
ever, higher nitrite concentrations are known to negatively
affect microorganisms due to its affinity for metal ions
which are attributable to the bacteriostatic property of
nitrite (Kroupova et al. 2018). Biofilms or microbial con-
sortium especially those of nitrifying bacteria have been
used for improving water quality by maintaining low levels
of total ammonia nitrogen, nitrite nitrogen and pathogenic
bacteria in culture systems (Yusoff et al. 2006; Shariff et al.
2009; Barik et al. 2018). The use of probiotic and nitrifying
bacterial consortium increased the survival and reduced the
ammonia and nitrite toxicity in experimental shrimp cul-
ture (Karthik et al. 2015). Supplementation of Bacillus
licheniformis as a water probiotic resulted in nitrite removal
and alterations of the microbial composition in grass carp
rearing system (Liang et al. 2015). Commercially available
probiotics such as Environ-AC (Wockard Biostate Ltd),
PROFS (Biostadt India Ltd.) and CIBAMOX (ICAR-CIBA)
15
A. Ciji and M. S. Akhtar
were found to be effective in maintaining good water qual-
ity including low levels of ammonia and nitrite in different
aquaculture systems (Lakshmanan & Soundarapandian
2008; Barman et al. 2015). Nitrifying bacteria Nitrosomonas
sp. and Nitrobacter sp attached to bentonite, zeolite, CaCO3
and amberlite in RAS had higher activity than free-living
strains as they utilize carbonate and bicarbonate alkalinity
as a nutrient source and help for maintenance of pH buffer-
ing capacity in the nitrification process (Diab & Shilo 1988;
Hao et al. 2017). Water probiotics can also be used as an
inoculum or start-up culture for lowering the time required
for the development of new or damaged biological filters
(Hochheimer & Wheaton 1991) and enhancing nitrifica-
tion efficiency of already established filters in RAS (Gross
et al. 2003). Some of the leading companies manufacturing
different types of biofilters and other equipments for com-
mercial scale recirculatory aquaculture system are AKVA
Group (https://www.akvagroup.com), Billund Aquaculture
(https://www.billundaquaculture.com), CM Aqua Tech-
nologies (https://www.cmaqua.dk), OxyGuard Interna-
tional A/S (http://www.oxyguard.dk), Hydrotech Inc.
(https://www.hydrotech.com) and Frea Aquaculture Solu-
tions (https://www.frea-solutions.com). The application of
biofloc technology in aquaculture is another promising
strategy to improve water quality. In addition to reducing
the demand for feed protein, the colonization of different
bacteria, algae, fungi and detritus in the biofloc assists in
the removal of ammonia and nitrite (Burford et al. 2004;
Holl et al. 2006; Serfling 2006). Moreover, biofloc technol-
ogy is one of the approaches to reduce the nitrous oxide
emissions associated with nitrification and denitrification
process from aquaculture systems (Hu et al., 2012). How-
ever, the potential of biofloc to increase the production of
carbon dioxide while minimizing N2O emissions should be
considered.
Micro- and macroalgae for nitrite removal
Micro/macroalgae either alone or in consortia with other
microorganisms, especially bacteria, can effectively remove
nitrogenous compounds from aquaculture systems as well
as from aquaculture effluent/wastewater (Jia & Yuan 2016).
Microalgae are photosynthetic organisms producing large
amounts of oxygen that can drive aerobic oxidation of
ammonia to low toxic nitrate. Furthermore, they can also
digest inorganic nitrogen sources such as ammonium,
nitrite and nitrate with ammonium as the most preferred
source and thereby help in the assimilation of these toxic
nitrogenous compounds (Zhou 2014; Jia & Yuan 2016).
Microalgae such as Chlorella sp., Scenedesmus, Chlorococcus
sp., Spirulina sp., Chlamydomonas and Phormidium are
most commonly employed for the removal of nitrogenous
compounds (Su et al. 2012; Cai et al. 2013; Alcantara et al.
16
2015; Jia & Yuan 2016). In algae–bacteria consortia, the
microalgae not only uptake nutrients but also provide dis-
solved oxygen for bacteria to remove nutrients (Karya et al.
2013). Wolfaardt et al. (1994) reported a 37% increase in
the efficiency of bacteria in the presence of microalgae.
Macroalgae (seaweeds) such as red algae of the genus
Gracilaria and Porphyra and green algae of the genus Ulva
have also been widely used as biofilters for the efficient
removal of nitrogenous compounds (Harlin et al. 1979;
Nelson et al. 2001; Msuya et al. 2006; Zhou et al. 2006;
Blouin et al. 2007).
Aquaponics recirculation system (ARS)/
Aquaculture-aquaponic systems
The integration of aquaculture with hydroponic cultivation
of plants is a viable alternative strategy not only to reduce
the nitrogenous compounds in RAS but also to reduce the
emission of green house gas, nitrous oxide from aquacul-
ture systems (Hu et al., 2012). The basic principle of
aquaponics system is to recycle the nutrient-rich wastewa-
ter from the aquaculture system in which plants act as a
natural filter as they uptake nutrients preventing its accu-
mulation and helps to maintain water quality (Rakocy
et al., 2006). Wide variety of plants including water spinach
(Ipomoea aquatica), mustard green (Brassica juncea) and
lettuce (Lactuca sativa) and the herbaceous plants, such as
basil (Ocimum basilicum), peppermint (Mentha x piperita)
and spearmint (Mentha spicata) can be grown in aquapon-
ics. Endut et al. (2011) evaluated the performance of water
spinach (Ipomoea aquatica) and mustard green (Brassica
juncea) in removing inorganic nitrogen and phosphate
from aquaculture wastewater and found that water spinach
was able to significantly reduce the total ammonia nitrogen,
nitrite-N, nitrate-N and orthophosphate with efficiencies of
78.32–85.48%, 82.93–92.22%, 79.17–87.10% and 75.36–
84.94%, respectively, compared to mustard green that
removed the nutrients in the range of 69.0–75.85% for total
ammonia nitrogen, 72.49–79.34% for nitrite-N, 66.67–
80.65% for nitrate-N and 66.79–77.87% for orthophos-
phate. However, the plants in the ARS can also contribute
to N2O emissions; hence, further research is needed to eval-
uate N2O emissions from aquaculture-aquaponic systems
for its further advocation.
Nutritional manipulations/interventions
The development of better nutritional strategies, particu-
larly the dietary incorporation of antioxidants, could be an
effective strategy to reverse the deleterious effects of higher
levels of nitrite (Ciji et al. 2013a). Although the ameliora-
tive effect of antioxidants on nitrite has long been recog-
nized, comparatively little experimental evidence for the
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 Problems and management of nitrite in aquaculture

role of dietary antioxidants in the toxicity of nitrite is avail-
able, especially in fish. In mammalian models, it is revealed
that the adverse effects of nitrite can be significantly
reduced by the addition of ascorbic acid or a-tocopherol
(Ames 1983; Hirneth & Classen 1984; Wagner et al. 1985;
Mirvish 1995). Intravenous administration of ascorbic acid
reduces methaemoglobin levels in rainbow trout (Cameron
1971). Reduced methaemoglobin levels (Fig. 7) were
observed in steelhead trout Salmo gairdneri and channel
catfish Ictalurus punctatus fed higher levels of ascorbic acid
(Blanco & Meade 1980; Wise et al. 1988). Recent studies
reported the protective role of vitamin E in ameliorating
the adverse effects of nitrite (Ciji et al. 2013a,2013c)
through enhanced activity of methaemoglobin reductase
and lowered tissue nitrite accumulation (Table 5). The
Figure 7 Dietary ascorbic acid (Vitamin C) reduces nitrite-induced
methaemoglobin levels in (i) Channel catfish when fed for 3 weeks and
then exposed to nitrite (Wise et al. 2010) and in (ii) Steelhead trout
when fed for 4 weeks and then exposed to nitrite (Blanco & Meade
1980). , Channel catfish (5 mg/L nitrite exposure); , Rainbow
trout (10 mg/L nitrite exposure)
exposure to environmental stressors or contaminants
including nitrite can potentiate the intracellular formation
of reactive species of oxygen (ROS), such as hydrogen per-
oxide, superoxide and the hydroxyl radical, and reactive
nitrogen species (RNS) capable of causing oxidative dam-
age to biological systems (Livingstone 2001; Monserrat
et al. 2007; Jensen & Hansen, 2011). The superoxide may
also interact readily with nitric oxide (NO) to form another
reactive and potent cytotoxic free radical peroxynitrite
(ONOO_), which is comparatively more stable than NO
(Pryor & Squadrito 1995; Van Dyke 1997; Squadrito &
Pryor 1998). The ability of ONOO_/ to cross cell mem-
branes contributes to its toxicity by allowing access to
intracellular target molecules (Macfadyen et al. 1999).
Nitric oxides and peroxynitrites can inhibit/inactivate sev-
eral enzyme systems such as ornithine decarboxylase (Seidel
et al. 2001), catalase (Keng et al. 2000), cytochrome
P450BM-3 (Daiber et al. 2000) and GPx (Padmaja et al.
1998). Beharka et al. (2002) have shown that vitamin E
supplementation reduces NO production in endotoxin-
stimulated macrophages of old mice. These findings suggest
that vitamin E is capable of reducing the production and/
or availability of not only superoxide but also of NO and
ONOO_/ and helps to attenuate oxidative damage resulting
from ROS (Shklar & Schwarz 1996; Klann et al. 1998; Azzi
et al. 1998; Chow & Hong 2002). It has been reported that
vitamin E supplementation reduces mortality and nitrite
toxicity-induced histopathological alterations of liver tis-
sues in rats (Hong & Chow 1987, 1988). By reducing avail-
able superoxide and NO, vitamin E may alleviate nitrite
toxicity by reducing the formation of reactive ONOO_/
(Chow & Hong 2002). However, it is not yet clear if the
action of vitamin E to reduce the generation of superoxide
and other ROS is independent of its antioxidant function.
In our earlier study, we noticed that dietary fortification of
vitamin E help to overcome the adverse effects of nitrite
exposure in Labeo rohita through enhanced activity of
methaemoglobin reductase and reduced formation of

Table 5 Effect of dietary L-tryptophan and vitamin E in alleviating nitrite toxicity with respect to the following physiological parameters in Labeo
rohita

Parameters Effect Dose-dependent trend

Vitamin E† L-tryptophan‡

Feed conversion ratio Reducing Y =
0.2x + 2.52, R2 = 0.892 Y =
0.09x + 2.433, R2 = 0.906
Specific growth rate Increasing Y = 0.045x + 0.346, R2 = 0.832 Y = 0.02x + 0.363, R2 = 0.923
Serum cortisol Reducing/ameliorating Y =
41.5x + 201.3, R2 = 0.847 Y =
36.5x + 198, R2 = 0.860
Methaemoglobin reductase Increasing Y = 0.006x + 0.017, R2 = 0.998 Y = 0.000x + 0.023, R2 = 0.75
Nitrite accumulation in liver Reducing Y =
0.125x + 2.14, R2 = 0.956 No definite trend

Source of data for relationship calculation:

†
Ciji et al. (2013a, 2014),
‡
Ciji et al. (2015)

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© 2019 Wiley Publishing Asia Pty Ltd 17
A. Ciji and M. S. Akhtar
superoxide, NO and peroxynitrite (Ciji et al. 2013a, 2014).
We further noticed that vitamin E aid to reverse the nitrite-
induced inhibition of steroidogenesis in Labeo rohita (Ciji
et al. 2013c). Selenium is another antioxidant that can be
used to reduce the deleterious effect of higher nitrite (Chow
& Hong 2002). Being a central part of glutathione peroxi-
dases (GPx), the enzyme responsible for the reduction in
organic hydroperoxides and H2O2 (Chow & Tappel 1974),
selenium acts synergistically with vitamin E in antioxidant
defence. In nutshell, dietary vitamin E/vitamin C reduces
the production and availability of superoxide and NO.while
selenium increases the level of seleno-enzymes/compounds
capable of scavenging ONOO_/ formed, if any, during
nitrite exposure and protect the fish from oxidative damage
and associated cell injuries (Sies et al. 1997; Chow & Hong
2002). However, Wang et al., (2006a,2006b) viewed that
higher dietary selenium is harmful as it increases the toxic-
ity of nitrite to prawn Penaeus vannamei. Dietary adminis-
tration of bovine lactoferrin (Fig. 8) is also found to be
promising in reducing the deleterious effect of nitrite in
M. rosenbergii (Chand et al. 2006).
Another potential dietary nutrient that can be used to
reduce the negative effect of nitrite intoxication is L-trypto-
phan (Table 5). Ciji et al. (2015) reported that dietary forti-
fication with higher levels of tryptophan found to be
effective in combating the nitrite-induced stress and trig-
gering the immunity in Labeo rohita. The authors further
suggested that the antioxidant properties of tryptophan
and its various metabolites (Keithahn & Lerchl 2005;
Reyes-Gonzales et al. 2009) as well as the potential role of
tryptophan in reducing the nitrite stress-induced elevation
of plasma cortisol might have protected L. rohita from the
adverse effect of nitrite. Recent studies in yellow catfish Pel-
teobagrus fulvidraco and crucian carp Carassius auratus
showed that exogenous supplementation of taurine, an
organic acid, helped to mitigate the adverse effect of high
Figure 8 Dietary administration of bovine lactoferine for 14 days
reduces percentage mortality due to nitrite (21 mg/L sodium nitrite)
stress in Macrobrachium rosenbergii (Chand et al. 2006). , Mortality;
, Linear (Mortality)
18
levels of ammonia by improving the antioxidative compe-
tence, nonspecific immunity and survival (Li et al. 2016;
Ren et al. 2016). The antioxidant property of shrimp caro-
tenoids, particularly b-carotene, astaxanthin and its esters,
has the protective effect against ammonia-induced stress in
characids Hyphessobrycon eques and common carp Cyprinus
carpio (Pan et al. 2011; Rama & Manjabhat 2014) as well as
protect post-larvae of shrimp Pleoticus muelleri against
nitrite stress (Diaz et al. 2014). However, the ameliorative
effect of taurine and shrimp carotenoids on nitrite toxicity
in fish needs to be explored elaborately.
Modification of pond bottom
The humic substances (HSs), which accumulate along with
toxic nitrogenous compounds in fish rearing systems, have
the potential to bind ammonia or nitrite due to the pres-
ence of carboxylic and amino acid functional groups in
them (Meinelt et al. 2010). Furthermore, the weak pH-buf-
fer capacity of HSs may stabilize the pH and, hence, prevent
the formation of toxic components (Meinelt et al. 2010). In
addition to its ability to detoxify nitrite and unionized
ammonia, the HSs have the capability to improve the stress
resistance of fish (Bouchnak & Steinberg 2010; Steinberg
et al. 2010; Meinelt et al. 2010). It has been suggested that
HSs can also interfere with the uptake and excretion of
nitrogenous compounds, thus decreasing their toxicity
(Meinelt et al. 2001; Matsuo & Val 2007). Hence, the addi-
tion of low-cost commercial humic substances (HSs) of
high reactivity to recirculating aquaculture systems is rec-
ommended as an alternative to chloride in reducing the
adverse effect of nitrite (Meinelt et al. 2010).
Ozonation of process water in recirculatory aquaculture
systems (RAS)
The high levels of water reuse in RAS generally lead to
accumulation of toxic nitrogenous compounds. Ozone,
popularly known as an effective disinfection agent, has the
potential to efficiently remove nitrite (Colberg & Lingg
1978; Rosenthal & Otte 1979; Schroeder et al. 2011) and
considerably improve the water quality in RAS without the
formation of any toxic ozone-produced oxidants (Summer-
felt & Hochheimer 1997; Summerfelt 2003; Tango & Gag-
non 2003). Ozone (O3) reacts instantly with nitrite and
converts it into low toxic nitrate with a rapid and constant
reaction rate of 3.7 9 105 M s
1 (Hoigne et al. 1985; Lin
& Wu 1996a,1996b). A quick nitrite oxidation by means of
instant ozonation is advantageous to sustain the intermedi-
ate step of nitrification in a biofilter, as this step is very sen-
sitive to changes in pH, organic load, substrate
concentration and other environmental changes (Kr€ uner &
Rosenthal 1983). As the process of ozonation and
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© 2019 Wiley Publishing Asia Pty Ltd

biofiltration for nitrite oxidation is competitive in nature,
batch ozonation is generally recommended for nitrite
removal (Schroeder et al. 2011). It has been suggested that
dietary administration of 3–25 g O3 per kg of feed to RAS
improved water quality and growth rate of fish (Summer-
felt et al. 2009; Martins et al. 2010). Ozonation is one of the
best suitable methods to reduce the nitrite levels in RAS
and fish/shellfish hatcheries.
Conclusion
This review reports the physiological consequences of
nitrite with greater emphasis on different methods/ap-
proaches for its mitigation/management in fish. Since
nitrite is a disrupter of multiple physiological functions
including ion regulatory, respiratory, cardiovascular,
endocrine and excretory processes, it is imperative to
monitor the water nitrite level regularly to enhance aqua-
culture production. The chances of nitrite problem can
be significantly reduced by avoiding overstocking and
overfeeding. Regular water exchange along with frequent
aeration and regular water testing can lessen the nitrite
problem to a greater extent. Newly constructed ponds
should be stocked gradually according to the efficiency of
biological filters to maintain the optimum water quality.
Addition of salt/chloride has been proven to be effective
in reducing the nitrite uptake but at the cost of dis-
turbing ionic homoeostasis. Adopting better nutritional
approach like feeding antioxidant-fortified diets could be
another effective and easy strategy to quash the adverse
impacts of nitrite toxicity in fish. Coupling these
approaches with an effectively functioning biological filter
will be helpful in dealing with nitrite issues in recircula-
tory aquaculture systems for sustainable production. Bio-
logical mitigation such as micro algae and biofloc
technology and electrochemical oxidation are promising
methods for nitrite removal. However, their applications
in real aquaculture are limited, and hence, more research
needs to be done in this area so that it can be explored
on commercial scale for better management of nitrite in
intensified aquaculture systems.
References
Abuzaid NS, Al-Hamouz Z, Bukhari AA, Essa MH (1999) Elec-
trochemical treatment of nitrite using stainless steel elec-
trodes. Water, Air and Soil Pollution 109: 429–442.
Aggergaard S, Jensen FB (2001) Cardiovascular changes and
physiological response during nitrite exposure in rainbow
trout. Journal of Fish Biology 59: 13–27.
Alcantara C, Mu~noz R, Norvill Z, Plouviez M, Guieysse B (2015)
Nitrous oxide emissions from high rate algal ponds treating
domestic wastewater. Bioresource Technology 177: 110–117.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Almendras JME (1987) Acute nitrite toxicity and methe-
moglobinemia in juvenile milkfish (Chanos chanos Forsskal).
Aquaculture 61: 33–40.
Alvarado NE, Quesada I, Hylland K, Marig omez I, Soto M
(2006) Quantitative changes in metallothionein expression in
target cell-types in the gills of turbot (Scophthalmus maximus)
exposed to Cd, Cu, Zn and after a depuration treatment.
Aquatic Toxicology 77: 64–77.
Ames BN (1983) Dietary carcinogens and anticarcinogens: oxy-
gen radicals and degenerative diseases. Science 221: 1256–
1264.
Apel K, Hirt H (2004) Reactive oxygen species: metabolism,
oxidative stress, and signal transduction. Annual Reviews in
Plant Biology 55: 373–399.
Arillo A, Gaino E, Margiocco C, Mensi P, Schenome G (1984)
Biochemical and ultrastructural effects of nitrite in rainbow
trout: Liver hypoxia as of the acute toxicity mechanism. Envi-
ronmental Research 34: 135–154.
Atwood HL, Fontenot QC, Tomasso JR, Isely JJ (2001) Toxicity
of nitrite to Nile tilapia: effect of fish size and environmental
chloride. North American Journal of Aquaculture 63: 49–51.
Auffret M, Yergeau E, Pilote A, Proulx E, Proulx D, Greer CW
et al. (2013) Impact of water quality on the bacterial popula-
tions and off-flavours in recirculating aquaculture systems.
FEMS Microbiology Ecology 84: 235–247.
Avilez IM, Altran AE, Aguiar LH, Moraes G (2004) Haematolog-
ical responses of the Neotropical teleost matrinxa (Brycon
cephalus) to environmental nitrite. Comparative Biochemistry
and Physiology Part C Toxicology and Pharmacology 139: 135–
139.
Azzi A, Aratri E, Boscoboinik D, Clement S, Ozer NK, Ricciarelli
R et al. (1998) Molecular basis of alphatocopherol control of
smooth muscle cell proliferation. BioFactors 7: 3–14.
Barik P, Ram R, Haldar C, Vardia HK (2018) Study on nitrifying
bacteria as bioremediator of ammonia in simulated aquacul-
ture system. Journal of Entomology and Zoology Studies 6(3):
1200–1206.
Barman P, Bandyopadhyay P, Mondal KC, Mohapatra PKD
(2015) Water quality improvement of Penaeus monodon cul-
ture pond for higher productivity through bioremediation.
Acta Biologica Szegediensis 59: 169–177.
Bartlett F, Neumann D (1998) Sensitivity of brown trout alevins
(Salmo trutta L.) to nitrite at different chloride concentra-
tions. Bulletin of Environmental Contamination and Toxicology
60: 340–346.
Bath RN, Eddy FB (1980) Transport of nitrite across fish gills.
Journal of Experimental Zoology 214: 119–121.
Beharka AA, Wu D, Serafini M, Meydani SN (2002) Mechanism
of vitamin E inhibition of cyclooxygenase activity in macro-
phages from old mice: role of peroxynitrite. Free Radical Biol-
ogy and Medicine 32: 503–511.
Bianchini A, Wasielesky W Jr, Miranda KC (1996) Toxicity of
nitrogenous compounds to juveniles of flatfish Paralichthys
orbignyanus. Bulletin of Environmental Contamination and
Toxicology 56: 453–459.
19
A. Ciji and M. S. Akhtar
Biesterfeld S, Farmer G, Russell P, Figueroa L (2003) Effect of
alkalinity type and concentration on nitrifying biofilm activ-
ity. Water Environment Research 75: 196–204.
Blanco O, Meade T (1980) Effect of dietary ascorbic acid on sus-
ceptibility of steelhead trout (Salmo gairdneri) to nitrite toxic-
ity. Revista de Biologia Tropical 28: 91–107.
Blevins WT, Schrader KK, Saadoun I (1995) Comparative physi-
ology of geosmin production by Streptomyces halstedii and
Anabaena sp. Water Science and Technology 31: 127–133.
Blouin N, Xiugeng F, Peng J, Yarish C, Brawley SH (2007) Seed-
ing nets with neutral spores of the red alga Porphyra umbili-
calis (L.) Kutzing for use in integrated multi-trophic
aquaculture (IMTA). Aquaculture 270: 77–91.
Boggs BK, Botte GG (2009) On-board hydrogen storage and
production: an application of ammonia electrolysis. Journal of
Power Sources 192: 573–581.
Bopp SK, Abicht HK, Knauer K (2008) Copper-induced oxida-
tive stress in rainbow trout gill cells. Aquatic Toxicology 86:
197–204.
Bouchnak R, Steinberg CEW (2010) Modulation of longevity in
Daphnia magna by food quality and simultaneous exposure to
dissolved humic substances. Limnologica 40: 86–91.
Bowser PR, Falls WW, Vanzandt J, Collier N, Phillips JD (1983)
Methemoglobinemia in channel catfish: methods of preven-
tion. The Progressive Fish Culturist 45: 154–158.
Boyd CE, Tucker CS (1998) Pond Aquaculture Water Quality Man-
agement, p. 700. Kluwer Academic Publishing, Boston, MA.
Brown DA, McLeay DJ (1975) Effect of nitrite on methe-
moglobin and total hemoglobin of juvenile steelhead trout
(Salmo gairdneri). Journal of the Fisheries Research Board of
Canada 35: 822–827.
Brown MN, Briones A, Diana J, Raskin L (2013) Ammonia-oxi-
dizing archaea and nitrite-oxidizing nitrospiras in the biofilter
of a shrimp recirculating aquaculture system. FEMS Microbi-
ology Ecology 83: 17–25.
Burford MA, Thompson PJ, McIntosh RP, Bauman RH, Pearson
DC (2004) The contribution of flocculated material to
shrimp, Litopenaeus vannamei, nutrition in a high-intensity,
zero-exchange system. Aquaculture 232: 525–537.
Buric M, Blahovec J, Kouril J (2016) Feasibility of open recircu-
lating system in temperate climate – a case study. Aquaculture
Research 47: 1156–1167.
Cai T, Park SY, Li Y (2013) Nutrient recovery from wastewater
streams by microalgae: status and prospects. Renewable and
Sustainable Energy Reviews 19: 360–369.
Camargo JA, Alonso A (2006) Ecological and toxicological
effects of inorganic nitrogen pollution in aquatic ecosystems:
a global assessment—a review. Environment International 32:
831–849.
Camargo JA, Alonso A, Salamanca A (2005) Nitrate toxicity to
aquatic animals: a review with new data for freshwater inver-
tebrates. Chemosphere 58: 1255–1267.
Cameron JN (1971) Methemoglobin in erythrocytes of rainbow
trout. Comparative Biochemistry and Physiology Part A Molec-
ular and Integrative Physiology 40: 743–749.
20
Campos BR, Furtado PS, D’Incao F, Wasielesky W, Poersch LHS
(2013) Nitrogen compounds on food consumption of pink-
shrimp Farfantepenaeus brasiliensis. Ciencia Rural 43: 2202–
2207.
Chakrabarti MH, Saleem M, Irfan MF, Raza S, Hasan DB, Daud
WMAW (2011) Application of waste derived activated carbon
felt electrodes in minimizing NaCl use for electrochemical dis-
infection of water. International Journal of Electrochemical
Science 6: 4470–4480.
Chand RK, Sahoo PK (2006) Effect of nitrite on the immune
response of freshwater prawn Macrobrachium malcolmsonii
and its susceptibility to Aeromonas hydrophila. Aquaculture
258: 150–156.
Chand RK, Sahoo PK, Kumari J, Pillai BR, Mishra BK (2006)
Dietary administration of bovine lactoferrin influences the
immune ability of the giant freshwater prawn Macrobrachium
rosenbergii (de Man) and its resistance against Aeromonas
hydrophila infection and nitrite stress. Fish and Shellfish
Immunology 21: 119–129.
Chen G (2004) Electrochemical technologies in wastewater treat-
ment. Separation and Purification Technology 38: 11–41.
Chen JC, Chen SF (1992) Effects of nitrite on growth and molt-
ing of Penaeus monodon juveniles. Comparative Biochemistry
and Physiology Part C Toxicology and Pharmacology 101(3):
453–458.
Chen JC, Cheng SY (1993) Hemolymph PCO2, hemocyanin,
protein levels and urea excretions of Penaeus monodon
exposed to ambient ammonia. Aquatic Toxicology 27: 281–
292.
Chen JC, Cheng SY (1995) Hemolymph oxygen content, oxyhe-
mocyanin, protein levels and ammonia excretion in the
shrimp Penaeus monodon exposed to ambient nitrite. Journal
of Comparative Physiology Part B Biochemical Systems and
Environmental Physiology 164: 530–535.
Chen JC, Cheng SY (2000) Recovery of Penaeus monodon from
functional anemia after exposure to sublethal concentration
of nitrite at different pH levels. Aquatic Toxicology 50: 73–83.
Chen JC, Lee Y (1997) Effect of nitrite exposure on acid-base
balance, respiratory protein, and iron concentrations of giant
freshwater prawn Macrobrachium rosenbergii at low pH.
Archives of Environmental Contamination and Toxicology 45:
35–46.
Chen S, Ling J, Blancheton JP (2006) Nitrification kinetics of
biofilm as affected by water quality factors. Aquaculture Engi-
neering 34: 179–197.
Cheng CH, Yang FF, Ling RZ, Liao SA, Miao YT, Ye CX et al.
(2015) Effects of ammonia exposure on apoptosis, oxidative
stress and immune response in pufferfish (Takifugu obscurus).
Aquatic Toxicology 164: 61–71.
Cheng SY, Chen JC (1998) Effects of nitrite exposure on hemo-
lymph electrolyte, respiratory protein and free amino acid
levels and water content of Penaeus japonicus. Aquatic Toxicol-
ogy 44: 129–139.
Cheng SY, Chen JC (1999) Hemocyanin oxygen affinity, and the
fractionation of oxyhemocyanin and deoxyhemocyanin for
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd

Penaeus monodon exposed to elevated nitrite. Aquatic Toxicol-
ogy 45: 35–46.
Cheng SY, Chen JC (2002) Joint action of elevated ambient
nitrite and nitrate on hemolymph nitrogenous compounds
and nitrogen excretion of tiger shrimp Penaeus monodon.
Comparative Biochemistry and Physiology Part C Toxicology
and Pharmacology 131: 303–314.
Cheng SY, Shieh LW, Chen JC (2013) Changes in hemolymph
oxyhemocyanin, acid–base balance, and electrolytes in Marsu-
penaeus japonicus under combined ammonia and nitrite
stress. Aquatic Toxicology 130–131: 132–138.
Cheng W, Liu CH, Chen JC (2002) Effect of nitrite on interac-
tion between the giant freshwater prawn Macrobrachium
rosenbergii and its pathogen Lactococcus garvieae. Diseases of
Aquatic Organisms 50: 189–197.
Chow CK, Hong CB (2002) Dietary vitamin E and selenium
and toxicity of nitrite and nitrate. Toxicology 180: 195–
207.
Chow CK, Tappel AL (1974) Response of glutathione peroxidase
as a function of dietary selenium in rats. Journal of Nutrition
104: 444–451.
Ciji A, Sahu NP, Pal AK, Dasgupta S, Akhtar MS (2012) Alter-
ations in serum electrolytes, antioxidative enzymes and
haematological parameters of Labeo rohita on short-term
exposure to sublethal dose of nitrite. Fish Physiology and Bio-
chemistry 38: 1355–1365.
Ciji A, Sahu NP, Pal AK, Akhtar MS (2013a) Physiological
changes in Labeo rohita during nitrite exposure: detoxifica-
tion through dietary vitamin E. Comparative Biochemistry
and Physiology Part C Toxicology and Pharmacology 158:
122–129.
Ciji A, Sahu NP, Pal AK, Akhtar MS, Meena DK (2013b)
Hemato-immunological responses of nitrite exposed Labeo
rohita fed diets supplemented with vitamin E and L-trypto-
phan and challenged by Aeromonas hydrophila. The Israeli
Journal of Aquaculture – Bamidgeh 65: 889–897.
Ciji A, Sahu NP, Pal AK, Akhtar MS (2013c) Nitrite-induced
alterations in sex steroids and thyroid hormones of Labeo
rohita juveniles: effects of dietary vitamin E and ltryptophan.
Fish Physiology and Biochemistry 39: 1297–1307.
Ciji A, Sahu NP, Pal AK, Akhtar MS, Tincy V, Mishal P et al.
(2014) Effect of dietary vitamin E and nitrite exposure on
growth and metabolic variables of Labeo rohita juveniles.
National Academy Science Letters 37(2): 123–129.
Ciji A, Sahu NP, Pal AK, Akhtar MS (2015) Dietary L-trypto-
phan modulates growth and immuno-metabolic status of
Labeo rohita juveniles exposed to nitrite. Aquaculture Research
46: 2013–2024.
Colberg PJ, Lingg AJ (1978) Effect of ozonation on microbial
fish pathogens, ammonia, nitrate, nitrite, and BOD in simu-
lated reuse hatchery water. Journal of Fisheries Research Board
of Canada 35: 1290–1296.
Colt J, Ludwig R, Tchobanoglous G, Cech J (1981) The effects of
nitrite on the short-term growth and survival of channel cat-
fish, Ictalurus punctatus. Aquaculture 24: 111–122.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Colt J, Tchobanoglous G (1976) Evaluation of the short-term
toxicity of nitrogenous compounds to channel catfish Ictalu-
rus punctatus. Aquaculture 8: 209–224.
Crawford RE, Allen GH (1977) Seawater inhibition of nitrite
toxicity to chinook salmon. Transactions of the American Fish-
eries Society 106: 105–109.
Daiber A, Herold S, Schoneich C, Namgaladze D, Peterson JA,
Ullrich V (2000) Nitration and inactivation of cytochrome
P450BM-3 by peroxynitrite stopped-flow measurements
prove ferryl intermediates. European Journal of Biochemistry
267: 6729–6739.
Das PC, Ayyappan S, Das BK, Jena JK (2004b) Nitrite toxicity in
Indian major carps: sublethal effect on selected enzymes in
fingerlings of Catla catla, Labeo rohita and Cirrhinus mrigala.
Comparative Biochemistry and Physiology Part C Toxicology
and Pharmacology 138: 3–10.
Das PC, Ayyappan S, Jena JK, Das BK (2004a) Nitrite toxicity in
Cirrhinus mrigala (Ham): acute toxicity and sub lethal effect on
selected haematological parameters. Aquaculture 235: 633–644.
De Groef B, Decallonne BR, Van der Geyten S, Darras VM,
Bouillon R (2006) Perchlorate versus other environmental
sodium/iodide symporter inhibitors: potential thyroid-related
health effects. European Journal of Endocrinology 155: 17–25.
Deane EE, Woo NYS (2007) Impact of nitrite exposure on endo-
crine, osmoregulatory and cytoprotective functions in the
marine teleost Sparus sarba. Aquatic Toxicology 82: 85–93.
Del Punta K, Charreau EH, Pignataro OP (1996) Nitric oxide
inhibits Leydig cell steroidogenesis. Endocrinology 137: 5337–
5343.
Delaforge M, Piffeteau A, Boucher JL, Viger A (1995) Nitric
oxide formation during the cytochrome P-450-dependent
reductive metabolism of 18-nitro-oxyandrostenedione. Jour-
nal of Pharmacology and Experimental Therapeutics 274: 634–
640.
Dehler CE, Secombes CJ, Martin SAM (2017) Environmental
and physiological factors shape the gut microbiota of Atlantic
salmon parr (Salmo salar L.). Aquaculture 467: 149–157.
Di Meglio S, Tramontano F, Cimmino G, Jones R, Quesada P
(2004) Dual role for poly (ADP-ribose) polymerase-1 and-2
and poly (ADP-ribose) glycohydrolase as DNA-repair and
pro-apoptotic factors in rat germinal cells exposed to nitric
oxide donors. Biochimica et Biophysica Acta Molecular Cell
Research 1692: 35–44.
Diab S, Shilo M (1988) Effect of adhesion to particles on the sur-
vival and activity of Nitrosomonas sp. and Nitrobacter sp.
Archives of Microbiology 150: 387–393.
Diaz V, Ibanez R, Gomez P, Urtiaga AM, Ortiz I (2010) Kinetics
of electro-oxidation of ammonia-N, nitrites and COD from a
recirculating aquaculture saline water system using BDD
anodes. Water Research 45: 125–134.
Diaz AC, Velurtas SM, Espino ML, Fenucci JL (2014) Effect
of dietary astaxanthin on free radical scavenging capacity
and nitrite stress tolerance of postlarvae shrimp Pleoticus
muelleri. Journal of Agricultural and Food Chemistry 62:
12326–12331.
21
A. Ciji and M. S. Akhtar
Doblander C, Lackner R (1996) Metabolism and detoxification
of nitrite by trout hepatocytes. Biochimica et Biophysica Acta
1289: 270–274.
Doblander C, Lackner R (1997) Oxidation of nitrite to nitrate in
isolated erythrocytes: a possible mechanism for adaptation to
environmental nitrite. Canadian Journal of Fish and Aquatic
Sciences 54: 157–161.
Doman RK, Perez M, Donato NJ (1999) JNK and p53 stress sig-
naling cascades are altered in MCF-7 cells resistant to tumor
necrosis factor-mediated apoptosis. Journal of Interferon and
Cytokine Research 19: 261–269.
Duan YF, Zhang Y, Dong HB, Wang Y, Zhang JS (2017) Effects
of dietary poly-b-hydroxybutyrate (PHB) on microbiota com-
position and the mTOR signaling pathway in the intestines of
Litopenaeus vannamei. Journal of Microbiology 55: 946–954.
Duan YF, Liu Q, Wang Y, Zhang J, Xiong D (2018) Impairment
of intestine barrier function in Litopenaeus vannamei exposed
to ammonia and nitrite stress. Fish and Shellfish Immunology
78: 279–288.
Duangsawasdi M, Sripoomun C. (1981) Duangsawasdi M, Sri-
poomun C (1981) Acute toxicities of ammonia and nitrite to
Clarius batrachus and their interaction to chlorides. Report of
the programme for the development of pond management
techniques and disease control (DoF - UNDP/FAO THA/75/
012) National Inland Fisheries Institute Bangkok, Thailand.
THA/75/012/WP 27.
Eddy FB, Kunzlik PA, Bath RN (1983) Uptake and loss of nitrite
from the blood of rainbow trout, Salmo gairdneri Richardson,
and Atlantic salmon, Salmo salar L. in freshwater and dilute
sea water. Journal of Fish Biology 23: 105–116.
Edwards TM, Guillette LJ Jr (2007) Reproductive characteris-
tics of male mosquito fish (Gambusia holbrooki) from
nitrate-contaminated springs in Florida. Aquatic Toxicology
85: 40–47.
Edwards TM, Hamlin HJ (2018) Reproductive endocrinology of
environmental nitrate. General and Comparative Endocrinol-
ogy 265: 31–40.
Edwards TM, Miller HD, Guillette LJ (2006a) Water quality
influences reproduction in female mosquitofish (Gambusia
holbrooki) from eight Florida springs. Environmental Health
Perspectives 114(1): 69–75.
Edwards TM, Moore BC, Guillette LJ (2006b) Reproductive dys-
genesis in wildlife: a comparative view. International Journal
of Andrology 29: 109–120.
El-Gohary M, Awara WM, Nassar S, Hawas S (1999) Deltame-
thrin-induced testicular apoptosis in rats: the protective effect
of nitric oxide synthase inhibitor. Toxicology 132: 1–8.
Ellis RP, Parry H, Spicer JI, Hutchinson TH, Pipe RK, Widdi-
combe S (2011) Immunological function in marine inverte-
brates: Responses to environmental perturbation. Fish and
Shellfish Immunology 30: 1209–1222.
Emamjomeh MM, Sivakumar M (2009) Review of pollutants
removed by electrocoagulation and electrocoagulation/flota-
tion processes. Journal of Environmental Management 90:
1663–1679.
22
Endut A, Jusoh A, Ali N, Wan Nik WB (2011) Nutrient removal
from aquaculture wastewater by vegetable production in
aquaponics recirculation system. Desalination and Water
Treatment 32: 422–430.
Eskiocak S, Dundar C, Basoglu T, Altaner S (2005) The effects of
taking chronic nitrate by drinking water on thyroid functions
and morphology. Clinical and Experimental Medicine 5: 66–71.
Evans DH (1993) The Physiology of Fishes. CRC Press, Boca
Raton, FL.
FAO (2016) State of World Fisheries and Aquaculture: Contribut-
ing to Food Security and Nutrition for All, p. 266. Food and
Agricultural Organization of the United Nations (FAO),
Rome. Italy.
Fleming WR (1974) Electrolyte metabolism of teleosts including
calcified tissues. In: Florkin M, Scheer BT (eds.) Chemical
Zoology, Vol. VIII, pp. 486–508. Academic Press, New York,
NY.
Foley J, Haas D, Yuan Z, Lant P (2010) Nitrous oxide generation
in full scale biological nutrient removal wastewater treatment
plants. Water Research 44: 831–834.
Fontenot QC, Isely JJ, Tomasso JR (1999) Characterization and
inhibition of nitrite uptake in shortnose sturgeon fingerlings.
Journal of Aquatic Animal Health 11: 76–80.
Frances J, Allan GL, Nowak BF (1998) The effects of nitrite on
the short-term growth of silver perch (Bidyanus bidyanus).
Aquaculture 163: 63–72.
Freitag AR, Thayer LR, Leonetti C, Stapleton HM, Hamlin HJ
(2015) Effects of elevated nitrate on endocrine function in
Atlantic salmon, Salmo salar. Aquaculture 436: 8–12.
Furtado PS, Valenzuela MAJ, Rodriguez-Fuentes G, Campos BR,
Wasielesky W Jr, Gaxiola G (2016) Chronic effect of nitrite on
the rearing of the white shrimp Litopenaeus vannamei in two
salinities. Marine and Freshwater Behaviour and Physiology 49:
201–211.
Gaino E, Arillo A, Mensi P (1984) Involvement of the gill chlo-
ride cells of trout under acute nitrite intoxication. Compara-
tive Biochemistry and Physiology Part A Molecular and
Integrative Physiology 77: 611–617.
Gill TS, Pande J, Tewari H (1991) Haemopathological changes
associated with experimental adicarb poisoning in fish (Pun-
tius conchonius Ham.). Bulletin of Environmental Contamina-
tion and Toxicology 7: 628–633.
Grosell M, Jensen FB (2000) Uptake and effects of nitrite in the
marine teleost fish Platichyhysflesus. Aquatic Toxicology 50:
97–107.
Gross A, Nemirovsky A, Zilberg D, Khaimov A, Brenner A, Snir
E et al. (2003) Soil nitrifying enrichments as biofilter starters
in intensive recirculating saline water aquaculture. Aquacul-
ture 223: 51–62.
Guillette LJ, Edwards TM (2005) Is nitrate an ecologically rele-
vant endocrine disruptor in vertebrates? Integrative and Com-
parative Biology 45: 19–27.
Guo H, Xian JA, Li B, Ye CX, Wang AL, Miao YT et al. (2013)
Gene expression of apoptosis-related genes, stress protein and
antioxidant enzymes in hemocytes of white shrimp
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd

Litopenaeus vannamei under nitrite stress. Comparative Bio-
chemistry and Physiology Part C: Toxicology and Pharmacology
157(4): 366–371.
Guttman L, van Rijn J (2008) Identification of conditions under-
lying production of geosmin and 2-methylisoborneol in a
recirculating system. Aquaculture 279: 85–91.
Gutzmer MP, Tomasso JR (1985) Nitrite toxicity to the crayfish
Procambarus clarkii. Bulletin of Environmental Contamination
and Toxicology 34: 369–376.
Han S, Wang B, Wang M, Liu Q, Zhao W, Wang L (2017)
Effects of ammonia and nitrite accumulation on the survival
and growth performance of white shrimp Litopenaeus van-
namei. International Surgery Journal 14: 221–232.
Hao DM, Tuyet DTA, Nga HTT, Dat NT, Sinh LX, Lan TD
(2017) Inorganic nitrogen removal in tropically coastal aqua-
culture ponds using integrated bioremediation. Journal of
Marine Science and Technology 17: 320–330.
Harlin MM, Thorne-Miller B, Thursby BG (1979) Ammonium
uptake by Gracilaria sp. (Florideophycea) and Ulva lactuca
(Chlorophycea) in closed system fish culture. In: Proceeding of
International Seaweed Symposium IX, Science Press, Princeton,
pp. 285–293.
Harris RR, Coley S (1991) The effect of nitrite on chloride regu-
lation in the crayfish Pacifastacus leniusculus Dana (Crustecea:
Decapoda). Journal of Comparative Physiology Part B 161:
199–206.
Hasan MR, Macintosh DJ (1986) Effect of chloride concentra-
tion on the acute toxicity of nitrite to common carp, Cyprinus
carpio L., fry. Aquaculture and Fisheries Management 17: 19–
30.
Higgs DA, Fagerlund UHM, McBride JR, Dyes HM, Donaldson
EM (1977) Influence of combinations of bovine growth hor-
mone, 17a methyltestosterone and L-thyroxine on growth of
yearling coho salmon (Onchorhynchus kisutch). Canadian
Journal of Zoology 55(6): 1048–1056.
Hilmy AM, El-Domiaty NA, Wershana K (1987) Acute and
chronic toxicity of nitrite to Clarias lazera. Comparative Bio-
chemistry and Physiology Part C: Toxicology and Pharmacology
8: 247–253.
Hinton DE, Baumann PC, Gardner GC, Hawkins WE, Hen-
dricks JD, Murchelano RA et al. (1992) Histopathologic
biomarkers. In: Huggett RJ, Kimerly RA, Mehrle PM, Jr Berg-
man HL (eds) Biomarkers: Biochemical, Physiological and His-
tological Markers of Anthropogenic Stress, pp. 155–210. Lewis
Publishers, Chelsea, MI.
Hirneth H, Classen HG (1984) Inhibition of nitrate-induced
increase of plasma nitrite and ethemoglobinemia in rats by
simultaneous feeding of ascorbic acid or tocopherol.
Arzneimittel-Forschung 34: 988–991.
Hochachck PW, Somero GN (2002) Biochemical Adaptation, p.
466. Oxford University Press, New York, NY.
Hochheimer JN, Wheaton FW (1991) Understanding biofilters,
practical microbiology for ammonia removal in aquaculture.
Proceedings of the Aquaculture Symposium, 4 – 6 April 1991,
Cornell University, Ithaca, NY, USA, pp. 57–83.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Hofer R, Gatumu E (1994) Necrosis of trout retina (Oncor-
hynchus mykiss) after sublethal exposure to nitrite. Archives of
Environmental Contamination and Toxicology 26: 119–123.
Hoigne J, Bader H, Haag WR, Staehelin J (1985) Rate constants
of reactions of ozone with organic and inorganic compounds
in water. III. Inorganic compounds and radicals. Water
Research 19: 993–1004.
Holl CM, Tallamy CJ, Moss SM (2006) Varied microbes impor-
tant to recirculating aquaculture systems. Global Aquaculture
Advocate 9: 38–39.
Hong CB, Chow CK (1987) Prevention of eosinophilic enteritis
and eosinophilia in rats with selenium and vitamin E. Federa-
tion Proceedings 46: 1154A.
Hong CB, Chow CK (1988) Induction of eosinophilic enteritis
and eosinophilia in rats by vitamin E and selenium deficiency.
Experimental and Molecular Pathology 48: 182–192.
Hooper AB, Vannelli T, Bergmann DJ, Arciero DM (1997) Enzy-
mology of the oxidation of ammonia to nitrite by bacteria.
Antonie Van Leeuwenhoek International Journal of General and
Molecular Microbiology 71: 59–67.
Howgate P (2004) Tainting of farmed fish by geosmin and 2-
methylisoborneol: a review of sensory aspects and of uptake/
depuration. Aquaculture 234: 155–181.
Hu Z, Lee JW, Chandran K, Kim S, Khanal SK (2012) Nitrous
oxide (N2O) emission from aquaculture: a review. Environ-
mental Science and Technology 46: 6470–6480.
Huey DW, Beitinger TL (1982) A methemoglobin reductase sys-
tem in channel catfish Ictalurus punctatus. Canadian Journal
of Zoology 60: 1511–1513.
Huey DW, Beitinger TL, Wooten MC (1984) Nitrite induced
methaemoglobin formation and recovery in channel catfish
(Ictalurus punctatus) at three acclimation temperatures. Bul-
letin of Environmental Contamination and Toxicology 32: 674–
681.
Huey DW, Simco BA, Criswell DW (1980) Nitrite-induced
methemoglobin formation in channel catfish. Transactions of
the American Fisheries Society 109: 558–562.
Inadomi C, Murata H, Ihara Y, Goto S, Urata Y, Yodoi J et al.
(2012) Overexpression of glutaredoxin protects cardiomy-
ocytes against nitric oxide-induced apoptosis with suppress-
ing the S-nitrosylation of proteins and nuclear translocation
of GAPDH. Biochemical and Biophysical Research Communi-
cations 425: 656-661.
Iwama GK, Thomas PT, Forsyth RB, Vijayan MM (1998) Heat
shock protein expression in fish. Reviews in Fish Biology and
Fisheries 8: 35–56.
Iwama GK, Vijayan MM, Forsyth RB, Ackerman PA (1999) Heat
shock proteins and physiological stress in fish. American Zool-
ogist 39: 901–909.
Jensen FB (1990a) Nitrite and red cell function in carp: control
factors for nitrite entry, membrane potassium ion perme-
ation, oxygen affinity and met haemoglobin formation. Jour-
nal of Experimental Biology 152: 149–166.
Jensen FB (1990b) Sublethal physiological changes in freshwater
crayfish, Astacus astacus exposed to nitrite: haemolymph and
23
A. Ciji and M. S. Akhtar
muscle tissue electrolyte status, and haemolymph acid–base
balance and gas transport. Aquatic Toxicology 18: 51–60.
Jensen FB (1992) Influence of haemoglobin conformation,
nitrite and eicosanoids on Kq transport across the carp red
blood cell membrane. Journal of Experimental Biology 171:
349–371.
Jensen FB (1993) Influence of nucleoside triphosphates, inor-
ganic salts, NADH, catecholamines, and oxygen saturation on
nitrite-induced oxidation of rainbow trout haemoglobin. Fish
Physiology and Biochemistry 12: 111–117.
Jensen FB (1995) Uptake and effects of nitrite and nitrate in ani-
mals. In: Walsh PJ, Wright P (eds) Nitrogen Metabolism and
Excretion, pp. 289–303. CRC Press, Boca Raton, FL.
Jensen FB (1996a) Physiological effects of nitrite in teleosts and
crustaceans. In: Taylor EW (ed) Toxicology of Aquatic Pollu-
tion: Physiological, Cellular and Molecular Approaches, pp.
169–186. Cambridge University Press, Cambridge.
Jensen FB (1996b) Uptake, elimination and effects of nitrite and
nitrate in freshwater crayfish (Astacus astacus). Aquatic Toxi-
cology 34: 95–104.
Jensen FB (2003) Nitrite disrupts multiple physiological func-
tions in aquatic animals. Comparative Biochemistry and Physi-
ology Part A Molecular and Integrative Physiology 135: 9–24.
Jensen FB (2007) Nitric oxide formation from nitrite in zebra-
fish. Journal of Experimental Biology 210: 3387–3394.
Jensen FB, Andersen NA, Heisler N (1987) Effects of nitrite
exposure on blood respiratory properties, acid base and elec-
trolyte regulation in the carp (Cyprinus carpio). Journal of
Comparative Physiology Part B 157: 533–541.
Jensen FB, Gerber L, Hansen MN, Madsen SS (2015) Metabolic
fates and effects of nitrite in brown trout under normoxic and
hypoxic conditions: blood and tissue nitrite metabolism and
interactions with branchial NOS, Na+/K+-ATPase and hsp70
expression. Journal of Experimental Biology 218: 2015–2022.
Jensen FB, Hansen MN (2011) Differential uptake and metabo-
lism of nitrite in normoxic and hypoxic goldfish. Aquatic Tox-
icology 101: 318–325.
Jensen FB, Nielsen K (2018) Methemoglobin reductase activity
in intact fish red blood cells. Comparative Biochemistry and.
Physiology Part A: Molecular & Integrative Physiology 216: 14–
19.
Jia H, Yuan Q (2016) Removal of nitrogen from wastewater
using microalgae and microalgae–bacteria consortia. Cogent
Environmental Science 2: 1275089.
Jia R, Han C, Lei JL, Liu BL, Huang B, Huo HH et al. (2015)
Effects of nitrite exposure on haematological parameters,
oxidative stress and apoptosis in juvenile turbot (Scophthal-
mus maximus). Aquatic Toxicology 169: 1–9.
Jia R, Liu BL, Han C, Huang B, Lei JL (2016) The physiological
performance and immune response of juvenile turbot (Scoph-
thalmus maximus) to nitrite exposure. Comparative Biochem-
istry and Physiology Part C: Toxicology and Pharmacology 181–
182: 40–46.
Jiang Q, Dilixiati A, Zhang W, Li W, Wang Q, Zhao Y et al.
(2013) Effect of nitrite exposure on metabolic response in the
24
freshwater prawn Macrobrachium nipponense. Central Euro-
pean Journal of Biology 9: 86–91.
Johansson MW, Keyser P, Sritunnyalucksana K, Soderhall K
(2000) Crustacean haemocytes and haematopoiesis. Aquacul-
ture 191: 45–52.
Jorgensen JB, Mustafa T (1980a) The effect of hypoxia on carbohy-
drate metabolism in flounder (Platichthys flesus L.). I. Utilization
of glycogen and accumulation of glycolytic end products in vari-
ous tissues. Comparative Biochemistry and Physiology Part B Bio-
chemistry and Molecular Biology 67: 243–248.
Jorgensen JB, Mustafa T (1980b) The effect of hypoxia on carbo-
hydrate metabolism in flounder (Platichthys flesus L.). II. High
energy phosphate compounds and the role of glycolytic and
gluconeogenetic enzymes. Comparative Biochemistry and
Physiology Part B Biochemistry and Molecular Biology 67: 249–
256.
Kamstra A, Span JA, Weerd JHV (1997) The acute toxicity and
sublethal effects of nitrite on growth and feed utilization of
European eel, Anguilla anguilla (L.). Aquaculture Research 27:
903–911.
Karthik R, Pushpam AC, Chelvan Y, Vanitha MC (2015) Effi-
cacy of probiotic and nitrifier bacterial consortium for the
enhancement of Litopenaeus vannamei aquaculture. Interna-
tional Journal of Veterinary Science and Research 1: 029–034.
Karya N, van der Steen NP, Lens PNL (2013) Photooxygenation
to support nitrification in an algal–bacterial consortium treat-
ing artificial wastewater. Bioresource Technology 134: 244–250.
Keithahn C, Lerchl A (2005) 5-Hydroxytryptophan is a more
potent in vitro hydroxyl radical scavenger than melatonin or
vitamin C. Journal of Pineal Research 38: 62–66.
Keng T, Privalle CT, Gilkeson GS, Weinberg JB (2000) Peroxyni-
trite formation and decreased catalase activity in autoimmune
MRL-lpr/lpr mice. Molecular Medicine 6: 779–792.
Khatoon H, Yusoff FM, Banerjee S, Shariff M, Mohamed S
(2007) Use of periphytic cyanobacterium and mixed diatoms
coated substrate for improving water quality, survival and
growth of Penaeus monodon Fabricius postlarvae. Aquaculture
271: 196–205.
Kiese M (1974) Methaemoglobinaemia: A Comprehensive Trea-
tise, pp. 55–125. CRC Press, Cleveland, OH.
Kim JH, Dahms HU, Han KN (2013) Biomonitoring of the river
pufferfish, Takifugu obscurus in aquaculture at different rear-
ing densities using stress-related genes. Aquaculture Research
44: 1835–1846.
Kim JW, Kim JY, Lim LJ, Kim SK, Choi HS, Hur YB (2018)
Effects of waterborne nitrite on hematological parameters and
stress indicators in olive flounders, Paralichthys olivaceus,
raised in bio-floc and seawater. Chemosphere 209: 28–34.
Klann EM, Roberson ED, Knapp LT, Sweatt JD (1998) A role for
superoxide in protein kinase C activation and induction of
long-term potentiation. Journal of Biological Chemistry 273:
4516–4522.
Knudsen PK, Jensen FB (1997) Recovery from nitrite induced
methaemoglobinaemia and potassium balance disturbances in
carp. Fish Physiology and Biochemistry 16: 1–10.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd

Koo JG, Kim SG, Jee JH, Kim JM, Bai SC, Kang JC (2005) Effects
of ammonia and nitrite on survival, growth and moulting in
juvenile tiger crab, Orithyia sinica (Linnaeus). Aquaculture
Research 36: 79–85.
Kozak P, Machova J, Policar T (2005) The effect of chloride con-
tent in water on the toxicity of sodium nitrite for spiny-cheek
crayfish (Orconectes limosus Raf.). Bulletin Francais de la P^eche
et de la Pisciculture 376–377: 705–714.
Kroupova H, Machova J, Svobodova Z (2005) Nitrite influence
on fish: a review. Veterinary Medicine Czech 11: 461–471.
Kroupova H, Valentova O, Svobodova Z, Sauer P, Machova J
(2018) Toxic effects of nitrite on freshwater organisms: a
review. Reviews in Aquaculture 10: 525–542.
Kr€
uner G, Rosenthal H (1983) Efficiency of nitrification in trick-
ling filters using different substrates. Aquaculture Engineering
2: 49–67.
Kshirsagar M, Gupta AB, Gupta SK (1995) Aerobic denitrifica-
tion studies on activated sludge mixed with Thiosphaera pan-
totropha. Environmental Technology 16: 35–43.
Kuhn DD, Drahos DD, Marsh L, Flick GJ Jr (2010) Evaluation
of nitrifying bacteria product to improve nitrification efficacy
in recirculating aquaculture systems. Aquacultural Engineering
43: 78–82.
Kumaraguru AK, Beamish FWH (1986) Effect of permethrin
(NRDC-173) on the bioenergetics of rainbow trout, Salmo
gairdneri. Aquatic Toxicology 9: 47–58.
Lahti E, Harri M, Lindqvist OV (1985) Uptake and distribution
of radioiodine, and the effect of ambient nitrate, in some fish
species. Comparative Biochemistry and Physiology Part A
Molecular and Integrative Physiology 80: 337–342.
Lakshmanan R, Soundarapandian P (2008) Effect of commercial
probiotics on large scale culture of black tiger shrimp Penaeus
monodon (Fabricius). Research Journal of Microbiology 3: 198–
203.
Lee K, Kim SJ, Kim D, Jo SH, Lee SJ, Choi SY (2014) Prostaglan-
din modulates TLR3-induced cytokine expression in human
astroglioma cells. Brain Research 1589: 54–60.
Lee KS, Chung EH, Han JH, Sohn HD, Jin BR (2003) cDNA
cloning of a defender against apoptotic death 1(DAD 1)
homologue, responsive to external temperature stimulus from
the spider, Araneus ventricosus. Comparative Biochemistry and
Physiology Part B: Biochemistry and Molecular Biology 135:
117–123.
Lefevre S, Jensen FB, Huong D, Wang T, Phuong NT, Bayley M
(2011) Effects of nitrite exposure on functional haemoglobin
levels bimodal respiration, and swimming performance in the
facultative air-breathing fish Pangasianodon hypophthalmus.
Aquatic Toxicology 104: 86–93.
Leatherland JF (1994) Reflections on the thyroidology of fishes:
from molecules to humankind. Guelph Ichthyology Reviews 2:
1–67.
Lefevre S, Jensen FB, Huong Do TT, Wang T, Phuong NT, Bay-
ley M (2012) Haematological and ion regulatory effects of
nitrite in the air-breathing snakehead fish Channa striata.
Aquatic Toxicology 118–119: 48–53.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Lewis WM, Morris DP (1986) Toxicity of nitrite to fish: a
review. Transactions of the American Fisheries Society 115:
183–195.
Li M, Lai H, Li Q (2016) Effects of dietary taurine on growth,
immunity and hyperammonemia in juvenile yellow catfish
Pelteobagrus fulvidraco fed all-plant protein diets. Aquaculture
450: 349–355.
Liang Q, Zhang X, Lee KH, Wang Y, Yu K, Shen W et al. (2015)
Nitrogen removal and water microbiota in grass carp culture
following supplementation with Bacillus licheniformis BSK-4.
World Journal of Microbiology and Biotechnology 31: 1711–
1718.
Liao PB, Mayo RD (1974) Intensified fish culture combining
water recycling with pollution abatement. Aquaculture 3: 61–
85.
Lin SH, Wu CL (1996a) Electrochemical removal of nitrate and
ammonia for aquaculture. Water Research 30: 715–721.
Lin SH, Wu CL (1996b) Removal of nitrogenous compounds
from aqueous solution by ozonation and ion exchange. Water
Research 30: 1851–1857.
Lin W, Guo H, Li Y, Wang L, Zhang D, Hou J et al. (2018b) Sin-
gle and combined exposure of microcystin-LR and nitrite
results in reproductive endocrine disruption via hypothala-
mic-pituitary-gonadal-liver axis. Chemosphere, 211, 1137–
1146.
Lin Y, Miao LH, Pan WJ, Huang X, Dengu JM, Zhang WX et al.
(2018a) Effect of nitrite exposure on the antioxidant enzymes
and glutathione system in the liver of bighead carp, Aris-
tichthys nobilis. Fish and Shellfish Immunology 76: 126–132.
Liu Y, Zang Y, Long X, Shao H, Liu Z, Zhang Z et al. (2013)
High nitrite removal capacity of an aerobic denitrifier Kleb-
siella oxytoca DF-1 isolated from aquaculture ponds in coastal
mudflats. African Journal of Microbiology Research 7: 1527–
1534.
Livingstone DR (2001) Contaminant-stimulated reactive oxygen
species production and oxidative damage in aquatic organ-
isms. Marine Pollution Bulletin 42: 656–666.
Losordo TM, Timmons MB (1994) An introduction to water re-
use systems. In: Timmons MB, Losordo TM (eds) Aquaculture
Water Reuse Systems: Engineering Design and Management,
pp. 1–7. Elsevier, Amsterdam.
Lu N, Gao NY, Deng Y, Li QS (2009) Nitrite formation during
low pressure ultraviolet lamp irradiation of nitrate. Water
Science and Technology 60: 1393–1400.
Lue YH, Hikim APS, Wang C, Leung A, Swerdloff RS (2003)
Functional role of inducible nitric oxide synthase in the
induction of male germ cell apoptosis, regulation of sperm
number, and determination of testes size: evidence from null
mutant mice. Endocrinology 144: 3092–3100.
Luzio A, Monteiro SM, Fontaınhas-Fernandes AA, Pinto-Car-
nide O, Matos M, Coimbra AM (2013) Copper induced
upregulation of apoptosis related genes in zebrafish (Danio
rerio) gill. Aquatic Toxicology 128: 183–189.
Maag M, Vinther FP (1996) Nitrous oxide emission by nitrifica-
tion and denitrification in different soil types and at different
25
A. Ciji and M. S. Akhtar
soil moisture contents and temperatures. Applied Soil Ecology
4: 5–14.
Macfadyen AJ, Reiter C, Zhuang Y, Beckman JS (1999) A novel
superoxide dismutase-based trap for peroxynitrite used to
detect entry of peroxynitrite into erythrocyte ghosts. Chemical
Research in Toxicology 12: 223–229.
Maetz J (1971) Fish gills: mechanisms of salt transfer in fresh
water and sea water. Philosophical Transactions of the Royal
Society B Biological Sciences 262: 209–249.
Mallasen M, Valenti WC (2006) Effect of nitrite on larval devel-
opment of giant river prawn Macrobrachium rosenbergii.
Aquaculture 261: 1292–1298.
Margiocco C, Arillo A, Mensi P, Shenone G (1983) Nitrite
bioaccumulation in Salmo gairdneri and haematological con-
sequences. Aquatic Toxicology 3: 261–270.
Mari KH, Jo~ao PB, Robbert K, Dimitry YS, Mark CM (2012)
Unravelling the reasons for disproportion in the ratio of AOB
and NOB in aerobic granular sludge. Applied Microbiology
and Biotechnology 94: 1657–1666.
Martins CIM, Eding EH, Verdegem MCJ, Heinsbroek LTN, Sch-
neider O, Blancheton JP et al. (2010) New developments in
recirculating aquaculture systems in Europe: a perspective on
environmental sustainability. Aquaculture Engineering 43: 83–
93.
Matsche MA, Markin E, Donaldson E, Hengst A, Lazur A (2012)
Effect of chloride on nitrite induced methaemoglobinemia in
Atlantic sturgeon, Acipenser oxyrichus oxyrinchus (Mitchill).
Journal of Fish Diseases 35: 873–885.
Matsuo AYO, Val AL (2007) Acclimation to humic substances
prevents whole body sodium loss and stimulates branchial
uptake capacity in cardinal tetras Paracheirodon axelrodi
(Schultz) subjected to extremely low pH. Journal of Fish Biol-
ogy 20: 989–1000.
Mazik PM, Hinman HL, Winkleman DA, Klaine SJ, Simco BA,
Parker NC (1991) Influence of nitrite and chloride concentra-
tions on survival and hematological profiles of striped bass.
Transactions of the American Fisheries Society 120: 247–254.
McConnell R (1985) Toxicity of Nitrite to the Fathead Minnow.
Colorado Department of Health File Report, Denver.
Meade TL, Perrone SJ (1980) Effect of chloride ion concentra-
tion and pH on the transport of nitrite across the gill epithelia
of coho salmon. The Progressive Fish Culturist 42: 71–72.
Meinelt T, Kroupova H, St€ uber A, Rennert B, Wienke A, Stein-
berg CEW (2010) Can dissolved aquatic humic substances
reduce the toxicity of ammonia and nitrite in recirculating
aquaculture systems? Aquaculture 306: 378–383.
Meinelt T, Playle RC, Pietrock M, Burnison BK, Wienke A,
Steinberg C (2001) Interaction of cadmium toxicity in
embryos and larvae of zebrafish (Danio rerio) with calcium
and humic substances. Aquatic Toxicology 54: 205–215.
Miao LH, Lin Y, Pan WJ, Huang X, Ge XP, Zhou QL et al.
(2018) Comparative transcriptome analysis reveals the gene
expression profiling in bighead carp (Aristichthys nobilis) in
response to acute nitrite toxicity. Fish and Shellfish Immunol-
ogy 79: 244–255.
26
Mirvish SS (1995) Role of N-nitroso compounds (NOC) and N-
nitrosation in etiology of gastric, esophageal, nasopharyngeal
and bladder cancer and contribution to cancer of known
exposures to NOC. Cancer Letters 93: 17–48.
Monserrat JM, Martinez PE, Geracitano L, Amado LL, Martins
C, Pinho GLL et al. (2007) Pollution biomarkers in estuarine
animals: Critical review and new perspectives. Comparative
Biochemistry and Physiology Part C Toxicology and Pharmacol-
ogy 146: 221–34.
Mook WT, Chakrabarti MH, Aroua MK, Khan GMA, Ali BS,
Islam MS et al. (2012) Removal of total ammonia nitrogen
(TAN), nitrate and total organic carbon (TOC) from aquacul-
ture wastewater using electrochemical technology: a review.
Desalination 285: 1–13.
Moraes G, Avilez IM, Hori TSF (2006) Comparison between
Biochemical responses of the teleost Pacu and its hybrid Tam-
bacu (Piaractus mesopotamicus x Colossoma macropomum) to
short term nitrite exposure. Brazilian Journal of Biology 66(4):
1103–1108.
Moraes G, Cattony EB, Souza RHS (1998) Metabolic
responses of the teleost Hoplias malabaricus to high levels
of environmental nitrite. Revista Brasileira De Biologia 58
(1): 105–113.
Morris AL, Hamlin HJ, Francis-Floyd R, Sheppard BJ, Guillette
LJ (2011) Nitrate induced goiter in captive whitespotted bam-
boo sharks Chiloscyllium plagiosum. Journal of Aquatic Animal
Health 23: 92–99.
Msuya FE, Kyewalyanga S, Salum D (2006) The performance of
the seaweed Ulva reticulate as a biofilter in a low-tech, low-
cost, gravity generated water flow regime in Zanzibar, Tanza-
nia. Aquaculture 254: 284–292.
Mukhi S, Carr JA, Anderson TA, Patino R (2005) Novel
biomarkers of perchlorate exposure in zebrafish. Environmen-
tal Toxicology and Chemistry 24: 1107–1115.
Nelson SG, Glenn EP, Conn J (2001) Cultivation of Gracilaria
parvispora (Rhodophyta) in shrimp-farm effluent ditches and
floating cages in Hawaii: a two-phase polyculture system.
Aquaculture 193: 239–248.
Ole F, Maurizio B, Francesca C, Serena R, Scot W, Israel P
(2009) Intramolecular electron transfer in Pseudomonas
aeruginosa cd1 nitrite reductase: thermodynamics and kinet-
ics. Biophysics Journal 96: 2849–2856.
Padmaja S, Squadrito GL, Pryor WA (1998) Inactivation of glu-
tathione peroxidase by peroxynitrite. Archives of Biochemistry
and Biophysics 349: 1–6.
Palachek RM, Tomasso JR (1984a) Nitrite toxicity to fathead
minnows: effect of fish weight. Bulletin of Environmental Con-
tamination and Toxicology 32: 238–242.
Palachek RM, Tomasso JR (1984b) Toxicity of nitrite to channel
catfish (Ictalurus punctatus), tilapia (Tilapia aurea), and large-
mouth bass (Micropterus salmoides): evidence for a nitrite
exclusion mechanism. Canadian Journal of Fisheries and
Aquatic Sciences 41: 1739–1744.
Pan CH, Chien YH, Wang YJ (2011) Antioxidant defense to
ammonia stress of characins (Hyphessobrycon eques
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd

Steindachner) fed diets supplemented with carotenoids.
Aquaculture Nutrition 17: 258–266.
Panesar NS, Chan KW (2000) Decreased steroid hormone syn-
thesis from inorganic nitrite and nitrate: studies in vitro and
in vivo. Toxicology and Applied Pharmacology 169: 222–230.
Park IS, Lee J, Hur JW, Song YC, Na HC, Noh CH (2007) Acute
toxicity and sublethal effects of nitrite on selected hematologi-
cal parameters and tissues in dark-banded rockfish, Sebastes
inermis. Journal of World Aquaculture Society 38: 188–199.
Park JW, Piknova B, Huang P, Noguchi CT, Schechter AN
(2013) Effect of blood nitrite and nitrate levels on murine pla-
telet function. PLoS ONE 8(2): e55699.
Pathan TS, Shinde SE, Thete PB, Sonawane DL (2010)
Histopathology of liver and kidney of Rasbora daniconius
exposed to paper mill effluent. Journal of Biological Sciences 5:
389–394.
Paz J (1984) The Effects of Borderline Alkalinity on Nitrification in
Natural Water Systems, p. 125. Polytechnic Institute of New
York, New York, NY.
Peakman M, Senaldi G, Vergani D (1989) Review: assessment of
complement activation in clinical immunology laboratories: time
for reappraisal? Journal of Clinical Pathology 42: 1018–1025.
Pequeux A (1995) Osmotic regulation in crustaceans. Journal of
Crustacean Biology 15: 1–60.
Perrone SJ, Meade TL (1977) Protective effect of chloride on
nitrite toxicity to coho salmon (Oncorhynchus kisutch). Jour-
nal of Fisheries Research of Board Canada 34: 486–492.
Peter MCS (2007) Thyroid hormones and hydromineral regula-
tion during stress in fish. D.Sc. thesis, Radboud University of
Nijmegen, The Netherlands.
Petrilli P, Tonukari NJ (1997) PFDB: a protein families database
for Macintosh computers. The effectiveness of its organization
in searching for protein similarity. Journal of Protein Chem-
istry 16: 713–720.
Philips S, Laanbroek HJ, Verstraete W (2002) Origin, causes and
effects of increased nitrite concentrations in aquatic environ-
ments. Reviews in Environmental Science and Biotechnology 1:
115–141.
Pistekova V, Voslarova E, Svobodova Z (2005) Nitrite toxicity to
Danio rerio: effects of chloride concentrations during acclima-
tization and in toxicity tests. Acta Veterinaria Brno 74: 435–
440.
Potts WTW, Fleming WR (1970) The effects of prolactin and
divalent ions on the permeability to water of Fundulus kansae.
Journal of Experimental Biology 53: 317–327.
Power DM, Llewellyn L, Faustino M, Nowell MA, Bjornsson
BT, Einarsdottir IE et al. (2001) Thyroid hormones in
growth and development of fish. Comparative Biochemistry
and Physiology Part C: Toxicology and Pharmacology 130
(4): 447–459.
Poxton MG, Allhouse SB (1982) Water quality criteria for mar-
ine fisheries. Aquaculture Engineering 1: 153.
Pryor WA, Squadrito GL (1995) The chemistry of peroxynitrite:
a product from the reaction of nitric oxide with superoxide.
American Journal of Physiology 268: L699–L722.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Rakocy JE, Masser MP, Losordo TM (2006) Recirculating aqua-
culture tank production systems: aquaponics- integrating fish
and plant culture. SRAC Publication No. 454. Southern
Regional Aquaculture Center. Texas A & M University, Texas,
USA.
Rama S, Manjabhat SN (2014) Protective effect of shrimp caro-
tenoids against ammonia stress in common carp Cyprinus car-
pio. Ecotoxicology and Environmental Safety 107: 207–213.
Ramano N, Zeng C (2013) Toxic effects of ammonia, nitrite,
and nitrate to decapod crustaceans: a review on factors influ-
encing their toxicity, physiological consequences, and coping
mechanisms. Reviews in Fisheries Science 21(1): 1–21.
Ramdhani N, Kumari S, Bux F (2013) Distribution of nitro-
somonas-related ammonia-oxidizing bacteria and nitrobac-
ter-related nitrite-oxidizing bacteria in two full-scale
biological nutrient removal plants. Water Environment
Research 85: 374–381.
Ramirez-Rochin J, Frias-Espericueta MG, Fierro-Sanudo JF,
Alarcon-Silvas SG, Fregoso-Lopez MG, Paez-Osuna F (2017)
Acute toxicity of nitrite on white shrimp Litopenaeus van-
namei (Boone) juveniles in low-salinity water. Aquaculture
Research 48: 2337–2343.
Ren Q, Li M, Yuan L, Song M, Xing X, Shi G et al. (2016) Acute
ammonia toxicity in crucian carp Carassius auratus and
effects of taurine on hyperammonemia. Comparative Bio-
chemistry and Physiology, Part C Toxicology and Pharmacology
190: 9–14.
Reyes-Gonzales MC, Fuentes-Broto L, Martı0 nez-Balları0 n E,
Miana-Mena FJ, Berzosa C, Garcı0 a-Gil FA et al. (2009) Effects
of tryptophan and 5-hydroxytryptophan on the hepatic cell
membrane rigidity due to oxidative stress. Journal of Mem-
brane Biology 231: 93–99.
Romano N, Zeng C (2009) Subchronic exposure to nitrite,
potassium and their combination on survival, growth, total
haemocyte count and gill structure of juvenile blue swimmer
crabs. Portunus pelagicus. Ecotoxicology and Environmental
Safety 72: 1287–1295.
Robertson RF, Hammond A, Jauncey K, Beveridge MCM, Law-
ton LA (2006) An investigation into the occurrence ofgeosmin
responsible for earthy–musty taints in UK farmed rainbow
trout, Onchorhynchus mykiss. Aquaculture 259: 153–163.
Roeselers G, Mittge EK, Stephens WZ, Parichy DM, Cavanaugh
CM, Guillemin K et al. (2011) Evidence for a core gut micro-
biota in the zebrafish. ISME Journal 5: 1595–1608.
Roques JAC, Schram E, Spanings T, Schaik T, Abbink W, Boer-
rigter J et al. (2013) The impact of elevated water nitrite con-
centration on physiology, growth and feed intake of African
catfish Clarias gariepinus (Burchell 1822). Aquaculture
Research 46: 1–12.
Rosenthal H, Otte G (1979) Ozonation in an intensive fish culture
recycling system. Ozone-Science and Engineering 1: 319–327.
Rungrassamee W, Klanchui A, Maibunkaew S, Chaiyapechara S,
Jiravanichpaisal P, Karoonuthaisiri N (2014) Characterization
of intestinal bacteria in wild and domesticated adult black
tiger shrimp (Penaeus monodon). PLoS ONE 9: e91853.
27
A. Ciji and M. S. Akhtar
Russo RC, Smith CE, Thurston RV (1974) Acute toxicity of
nitrite to rainbow trout (Salmo gairdneri). Journal of the Fish-
eries Research Board of Canada 31: 1653–1655.
Russo RC, Thurston RV (1977) The acute toxicity of nitrite to
fishes. In: Tubb RA (ed) Recent Advances in Fish Toxicology: A
Symposium, pp. 118–131. United States Environmental Pro-
tection Agency, Corvallis.
Russo RC, Thurston RV, Emerson K (1981) Acute toxicity of
nitrite to rainbow trout (Salmo gairdneri): Effects of pH,
nitrite species and anion species. Canadian Journal of Fisheries
and Aquatic Sciences 38: 387–393.
Saleem M, Essa MH (2010) Suitability for sustainable reuse of
secondary effluent: a case study in Saudi Arabia. NED Univer-
sity Journal of Research 7: 23.
Sampaio LA, Wasielesky W, Miranda Filho KC (2002) Effect of
salinity on acute toxicity of ammonia and nitrite to juvenile
Mugil platanus. Bulletin of Environmental Contamination and
Toxicology 68: 668–674.
Saoud IP, Naamani S, Ghanawi J, Nasser N (2014) Effects of
acute and chronic nitrite exposure on rabbitfish Siganus rivu-
latus growth, hematological parameters, and gill histology.
Journal of Aquaculture Research and Development 5: 263–271.
Scarano G, Saroglia MG (1984) Recovery of fish from functional
and hemolytic anemia after brief exposure to a lethal concen-
tration of nitrite. Aquaculture 43: 421–426.
Schoore JE, Simco BA, Davis KB (1995) Responses of blue cat-
fish and channel catfish to environmental nitrite. Journal of
Aquatic Animal Health 7: 304–311.
Schrader KK, Davidson JW, Summerfelt ST (2013) Evaluation of
the impact of nitrate-nitrogen levels in recirculating aquacul-
ture systems on concentration of offflavor compounds geos-
min and 2-methylisoborneol in water and rainbow trout
(Oncorhynchus mykiss). Aquaculture Engineering 57: 126–130.
Schroeder JP, Croot PL, Von Dewitz B, Waller U, Hanel R
(2011) Potential and limitations of ozone for the removal of
ammonia, nitrite and yellow substances in marine recirculat-
ing aquaculture systems. Aquacultural Engineering 45: 35–41.
Schwedler TE, Tucker CS (1983) Empirical relationship between
percent methemoglobin in channel catfish and dissolved
nitrite and chloride in ponds. Transactions of the American
Fisheries Society 112(1): 117–119.
Seidel ER, Ragan V, Liu L (2001) Peroxynitrite inhibits the activ-
ity of ornithine decarboxylase. Life Sciences 68: 1477–1483.
Serfling SA (2006) Microbial flocs. Natural treatment method
supports freshwater, marine species in recirculating systems.
Global Aquaculture Advocate 9: 34–36.
Shan HW, Bao WY, Ma S, Wei DP, Gao L (2016) Ammonia and
nitrite nitrogen removal in shrimp culture by Vibrio algi-
nolyticus VZ5 immobilized in SA beads. Aquaculture Interna-
tional 24: 357–372.
Shariff M, Yusoff FM, Banerjee S (2009) Agent and system for
improving water quality. Patent filed, application No.
PI20097001.
Sharrer MJ, Summerfelt ST, Bullock GL, Gleason LE, Taeuber J
(2005) Inactivation of bacteria using ultraviolet irradiation in
28
a recirculating salmonid culture system. Aquacultural Engi-
neering 33: 135–149.
Shklar G, Schwarz JL (1996) Vitamin E inhibits experimental
carcinogenesis and tumor angiogenesis. European Journal of
Cancer Part B Oral Oncology 32: 114–119.
Sies H, Sharov VS, Klotz LO, Briviba K (1997) Glutathione per-
oxidase protects against peroxynitrite-mediated oxidations. A
new function for selenoproteins as peroxynitrite reductase.
Journal of Biological Chemistry 272: 27812–27817.
Siikavuopio SI, Saether B (2006) Effects of chronic nitrite expo-
sure on growth in juvenile Atlantic cod, Gadus morhua. Aqua-
culture 255: 351–356.
Siikavuopio SI, Dale T, Christiansen JS, Nevermo I (2004)
Effects of chronic nitrite exposure on gonad growth in green
sea urchin Strongylocentrotus droebachiensis. Aquaculture 242:
357–363.
Siwicki AK, Antychowicz J, Waluga J (1994) Choroby ryb hodow-
lanych, p. 376. IRS, Olsztyn.
Smith C, Williams WG (1974) Experimental nitrite toxicity in
rainbow trout and Chinook salmon. Transactions of the Amer-
ican Fisheries Society 103: 389–390.
Sowers AD, Young SP, Isely JJ, Browdy CL, Tomasso JR (2004)
Nitrite toxicity to Litopenaeus vannamei in water containing
low concentrations of sea salt or mixed salts. Journal of the
World Aquaculture Society 35: 445–451.
Squadrito GL, Pryor WA (1998) Oxidative chemistry on nitric
oxide: the roles of superoxide, peroxynitrite, and carbon diox-
ide. Free Radical Biology and Medicine 25: 392–403.
Steinberg CEW, Ouerghemmi N, Herrmann S, Bouchnak R,
Timofeyev MA, Menzel R (2010) Stress by poor food quality
and exposure to humic substances: Daphnia magna responds
with oxidative stress, lifespan extension, but reduced offspring
numbers. Hydrobiologia 652: 223–226.
Stormer J, Jensen FB, Rankin JC (1996) Uptake of nitrite,
nitrate, and bromide in rainbow trout, Oncorhynchus mykiss:
effects on ionic balance. Canadian Journal of Fisheries and
Aquatic Sciences 53: 1943–1950.
Stone NM, Thomforde HK (2004) Understanding your fish
pond water analysis report. Cooperative Extension Program,
University of Arkansas at Pine Bluff Aquaculture/Fisheries.
Su Y, Mennerich A, Urban B (2012) Comparison of nutrient
removal capacity and biomass settleability of four high-poten-
tial microalgal species. Bioresource Technology 124: 157–162.
Summerfelt ST (2003) Ozonation and UV irradiation – an intro-
duction and examples of current applications. Aquaculture
Engineering 28: 21–36.
Summerfelt ST, Hochheimer JN (1997) Review of ozone pro-
cesses and applications as an oxidizing agent in aquaculture.
Progressive Fish Culturist 59: 94–105.
Summerfelt ST, Sharrer MJ, Tsukuda SM, Gearheart M (2009)
Process requirements for achieving full- flow disinfection or
recirculating water using ozonation and UV irradiation.
Aquaculture Engineering 40: 17–27.
Sun S, Ge X, Zhu J, Xuan F, Jiang X (2014a) Identification and
mRNA expression of antioxidant enzyme genes associated
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd

with the oxidative stress response in the Wuchang bream
(Megalobrama amblycephala Yih) in response to acute nitrite
exposure. Comparative Biochemistry and Physiology Part C:
Toxicology and Pharmacology 159: 69–77.
Sun S, Zhu J, Ge X, Zhang C, Miao L, Jiang X (2014b) Cloning
and expression analysis of a heat shock protein 90 b isoform
gene from the gills of Wuchang bream (Megalobrama ambly-
cephala Yih) subjected to nitrite stress. Genetics and molecular
research 14: 3036–3051.
Svobodova Z, Machova J, Drastichova J, Groch L, Luskova V,
Poleszczuk G et al. (2005a) Haematological and biochemical
profile of carp blood following nitrite exposure at different
concentration of chloride. Aquaculture Research 36: 1177–
1184.
Svobodova Z, Machova J, Poleszczuk G, Huda J, Hamackova J,
Kroupova H (2005b) Nitrite poisoning of fish in aquaculture
facilities with water-recirculating systems: three case studies.
Acta Veterinaria Brno 74: 129–137.
Tacon AGJ, Cody JJ, Conquest LD, Divakaran S, Forster IP
(2002) Effect of culture system on the nutrition and growth
performance of Pacific white shrimp Litopenaeus vannamei
(Boone) fed different diets. Aquaculture nutrition 8(2): 121–
137.
Tango MS, Gagnon GA (2003) Impact of ozonation on water
quality in marine recirculation systems. Aquaculture Engineer-
ing 29: 125–137.
Thangam Y (2014) Histopathological Studies on Nitrite Tox-
icity to Freshwater Fish, Cirrhinus mrigala. IOSR Journal
of Environmental Science Toxicology and Food Technology 8:
10–14.
Thillart GV, Kesbeke F, Waarde AV (1980) Anaerobic energy-
metabolism of goldfish, Carassius auratus (L.). Journal of
Comparative Physiology 136: 45–52.
Tipsmark CK, Madsen SS (2003) Regulation of Na+/K+-ATPase
activity by nitric oxide in the kidney and gill of the brown
trout (Salmo trutta). Journal of Experimental Biology 206:
1503–1510.
Tomasso JR, Davis KB, Simco BA (1981) Plasma corticosteroid
dynamics in channel catfish (Ictalurus punctatus) exposed to
ammonia and nitrite. Canadian Journal of Fisheries and Aqua-
tic Sciences 38: 1106–1112.
Tomasso JR, Wright MI, Simco BA, Davis KB (1980) Inhibition
of nitrite induced toxicity in channel catfish by calcium chlo-
ride and sodium chloride. The Progressive Fish Culturist 42:
144–146.
Tomasso JR (2012) Environmental nitrite and aquaculture: a
perspective. Aquaculture International 20: 1107–1116.
Tonacchera M, Pinchera A, Dimida A, Ferrarini E, Agretti P,
Vitti P et al. (2004) Relative potencies and additivity of per-
chlorate, thiocyanate, nitrate, and iodide on the inhibition of
radioactive iodide uptake by the human sodium iodide sym-
porter. Thyroid 14: 1012–1019.
Tort L, Balasch J, Mackenzie S (2003) Fish immune system. A
crossroads between innate and adaptive responses.
Inmunologıa 22: 277–286.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Tseng IT, Chen JC (2004) The immune response of white
shrimp Litopenaeus vannamei and its susceptibility to Vibrio
alginolyticus under nitrite stress. Fish and Shellfish Immunol-
ogy 17(4): 325–333.
Tucker CS (2000) Off-flavor problems in aquaculture. Reviews
in Fisheries Science 8: 45–88.
Tucker CS, Hargreaves JA (2004) Pond water quality. In: Tucker
CS, Hargreaves JA (eds) Biology and Culture of Channel Cat-
fish, pp. 216–278. Elsevier, Amsterdam.
€
Uner € Canli M, Sevgler Y (2001) Effects of cyperme-
N, Orucß EO,
thrin on antioxidant enzyme activities and lipid peroxidation
in liver and kidney of the freshwater fish, Oreochromis niloti-
cus and Cyprinus carpio (L.). Bulletin of Environmental Con-
tamination and Toxicology 67: 657–664.
Van Breemen N, Van Dijk HFG (1988) Ecosystems effects of
atmospheric deposition of nitrogen in the Netherlands. Envi-
ronmental Pollution 54: 249–274.
Van Dyke K (1997) The possible role of peroxynitrite in Alzhei-
mer’s disease: a simple hypothesis that could be tested more
thoroughly. Medical Hypotheses 48: 375–389.
Van Rijn J, Tal Y, Schreier HJ (2006) Denitrification in recircu-
lating systems: theory and applications. Aquacultural Engi-
neering 34: 364–376.
Vedel NE, Korsgaard B, Jensen FB (1998) Isolated and combined
exposure to ammonia and nitrite in rainbow trout (Oncor-
hynchus mykiss): effect on electrolyte status, blood respiratory
properties and brain glutamine/glutamate concentrations.
Aquatic Toxicology 41: 325–342.
Viant M, Werner I, Rosenblum E, Gantner A, Tjeerdema R, John-
son M (2003) Correlation between heat-shock protein induction
and reduced metabolic condition in juvenile steelhead trout
(Oncorhynchus mykiss) chronically exposed to elevated tempera-
ture. Fish Physiology and Biochemistry 29: 159–171.
Voslarova E, Pistekova V, Svobodova Z (2006) Nitrite toxicity to
Danio rerio: effects of fish age and chloride concentrations.
Acta Veterinaria Brno 75: 107–113.
Wagner DA, Shuker DE, Bilmazes C, Obiedzinski M, Baker I,
Young VR et al. (1985) Effect of vitamins C and E on endoge-
nous synthesis of N-nitrosamino acids in humans: precursor-
product studies with [15N] nitrate. Cancer Research 45:
6519-/6522.
Wang H, Liu CF, Qin CX, Cao SQ, Ding J (2007) Using a
macroalgae Ulva pertusa biofilter in a recirculating system for
production of juvenile sea cucumber Apostichopus japonicus.
Aquacultural Engineering 36: 217–224.
Wang J, Tang H, Zhang X, Xue X, Zhu X, Chen Y et al. (2018)
Mitigation of nitrite toxicity by increased salinity is associated
with multiple physiological responses: a case study using an
economically important model species, the juvenile obscure
puffer (Takifugu obscurus). Environmental Pollution 232: 137–
145.
Wang WN, Wang AL, Zhang YJ (2006a) Effect of dietary higher
level of selenium and nitrite concentration on the cellular
defense response of Penaeus vannamei. Aquaculture 256: 558–
563.
29
A. Ciji and M. S. Akhtar
Wang WN, Wang AL, Zhang YJ, Li ZH, Wang JX, Sun RY
(2004) Effects of nitrite on lethal and immune response of
Macrobrachium nipponense. Aquaculture 232: 679–686.
Wang Y, Zhang W, Li W, Xu Z (2006b) Acute toxicity of nitrite
to tilapia (Oreochromis niloticus) at different external chloride
concentrations. Fish Physiology and Biochemistry 32: 49–54.
Wasielesky W (2000) Cultivo de juvenis do camarao-rosa
Farfantepenaeus paulensis (Decapoda, Penaeidae) no estu-
ario da Lagoa dos Patos: efeitos dos parametros ambientais
[Culture of pink shrimp juveniles Farfantepenaeus paulensis
(Decapoda, Penaeidae) in the estuary of the Patos Lagoon:
effects of environmental parameters]. Tese de doutorado:
Fundacao Universidade Federal de Rio Grande, Rio
Grande, RS, 199.
Wedemeyer GA, Yasutake WT (1978) Prevention and treat-
ment of nitrite toxicity in juvenile steelhead trout (Salmo
gairdneri). Journal of the Fisheries Research Board of Canada
35: 822–827.
Weirich CR, Tomasso JR, Smith TIJ (1993) Toxicity of ammonia
and nitrite to sunshine bass in selected environments. Journal
of Aquatic Animal Health 5: 64–72.
Weitzberg E, Lundberg JON (1998) Nonenzymatic nitric oxide
production in humans. Nitric Oxide Biology and Chemistry 2:
1–7.
Wickins JF (1976) The tolerance of warm-water prawn to recir-
culated water. Aquaculture 9: 19–37.
Williams J, Crutzen PJ (2010) Nitrous oxide from aquaculture.
Nature Geoscience 3: 143–143.
Williams EM, Eddy FB (1986) Chloride uptake in freshwater tel-
eosts and its relationship to nitrite uptake and toxicity. Jour-
nal of Comparative Physiology B 156: 867–872.
Williams EM, Eddy FB (1988) Anion transport, chloride cell
number and nitrite-induced methaemoglobinaemia in rain-
bow trout (Salmo gairdneri) and carp (Cyprinus carpio).
Aquatic Toxicology 13: 29–42.
Wise DJ, Tomasso JR, Brandt TM (1988) Ascorbic acid inhibi-
tion of nitrite induced methemoglobinemia in channel catfish.
Progressive Fish Culturist 50: 77–80.
Wolfaardt GM, Lawrence JR, Robarts RD, Caldwell DE (1994)
The role of interactions, sessile growth, and nutrient amend-
ments on the degradative efficiency of a microbial consor-
tium. Canadian Journal of Microbiology 40: 331–340.
Woo NYS, Chiu SF (1994) Effects of nitrite exposure on growth
and survival of sea bass, Lates calcarifer, fingerlings in different
salinities. Journal of Applied Aquaculture 4: 45–54.
Woo NYS, Chiu SF (1997) Metabolic and Osmoregulatory
responses of the sea bass Lates calcarifer to nitrite expo-
sure. Environmental Toxicology and Water Quality 12: 257–
264.
Woo NYS, Wu R (1984) Changes in biochemical composition in
the red grouper, Epinephelus akaara (Temminck and Sch-
legel), and the black sea bream, Mylio macrocephalus (Basi-
lewsky), during hypoxic exposure. Comparative Biochemistry
and Physiology Part A Molecular and Integrative Physiology 77:
475–482.
30
Wu RSS, Zhou BS, Randall DJ, Woo NYS, Lam PKS (2003)
Aquatic hypoxia is an endocrine disruptor and impairs fish
reproduction. Environmental Science and Technology 37:
1137–1141.
Wuertz S, Schulze S, Eberhardt U, Schulz C, Schroeder J (2013)
Acute and chronic nitrite toxicity in juvenile pike-perch (San-
der lucioperca) and its compensation by chloride. Comparative
Biochemistry and Physiology Part C: Toxicology and Pharmacol-
ogy 157: 352–360.
Xian JA, Wang AL, Chen XD, Gou NN, Miao YT, Liao SA et al.
(2011) Cytotoxicity of nitrite on haemocytes of the tiger
shrimp, Penaeus monodon, using flow cytometric analysis.
Aquaculture 317: 240–244.
Xian JA, Wang AL, Hao XM, Miao YT, Li B, Ye CX et al. (2002)
In vitro toxicity of nitrite on haemocytes of the tiger shrimp,
Penaeus monodon, using flow cytometric analysis. Compara-
tive Biochemistry and Physiology Part C: Toxicology and Phar-
macology 156: 75–79.
Xiong JB, Wang K, Wu JF, Qiuqian LL, Yang KJ, Qian YX et al.
(2015) Changes in intestinal bacterial communities are closely
associated with shrimp disease severity. Applied Microbiology
and Biotechnology 99: 6911–6919.
Yamada T, Takatsu Y, Kasumi M, Marubashi W, Ichimura K
(2004) A homolog of the defender against apoptotic death
gene (DAD 1) in senescing gladiolus petals is down regulated
prior to the onset of programmed cell death. Journal of Plant
Physiology 161: 1281–1283.
Yanbo W, Wenju Z, Li Weifen, Zirong Xu (2006) Acute toxicity
of nitrite on tilapia (Oreochromius niloticus) at different exter-
nal chloride concentrations. Fish Physiology and Biochemistry
32: 49–54.
Yin G, Li W, Lin Q, Lin X, Lin J, Zhu Q et al. (2014) Dietary
administration of laminarin improves the growth perfor-
mance and immune responses in Epinephelus coioides. Fish
and Shellfish Immunology 41: 402–406.
Yu KL, Woo NYS (1987) Metabolic adjustments of an air breathing
teleost, Channa maculata, to acute and prolonged exposure to
hypoxic water. Journal of Fish Biology 31: 165–175.
Yusoff FM, Shariff M, Suhaila M, Kuppan P (2006) Periphyton
based apparatus for improving aquaculture production and
methods of preparing thereof. Patent filed application No.
P120063949.
Zhang Y, Ye C, Wang A, Zhu X, Chen C, Xian J et al. (2015) Iso-
lated and combined exposure to ammonia and nitrite in giant
freshwater pawn (Macrobrachium rosenbergii): effects on the
oxidative stress, antioxidant enzymatic activities and apopto-
sis in haemocytes. Ecotoxicology 24: 1601–1610.
Zheng J, Mao Y, Su Y, Wang J (2016) Effects of nitrite stress on
mRNA expresson of antioxidant enzymes, immune-related
genes and apoptosis-related proteins in Marsupenaeus japoni-
cas. Fish and Shellfish Immunology 58: 239–252.
Zhou W (2014) Potential applications of microalgae in wastewa-
ter treatments. In: Liu J, Sun Z, Gerken H (eds) Recent
Advances in Microalgal Biotechnology, pp. 1–9. OMICS Group
ebook, Foster City, CA.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd

Zhou Y, Yang HS, Hu HY, Liu Y, Mao YZ, Zhou H et al. (2006)
Bioremediation potential of the macroalga Gracilaria lemanei-
formis (Rhodophyta) integrated into fed fish culture in coastal
waters of north China. Aquaculture 252: 264–276.
Zhu L, Song L, Zhang H, Zhao J, Li C, Xu W (2008) Molecular
cloning and responsive expression to injury stimulus of a
defender against apoptotic death 1(DAD 1) gene from bay
scallops Argopecten irradians. Molecular Biology Reports 35:
125–132.
Reviews in Aquaculture, 1–31
© 2019 Wiley Publishing Asia Pty Ltd
Problems and management of nitrite in aquaculture
Zini A, Obryan MK, Magid MS, Schlegel PN (1996) Immuno-
histochemical localization of endothelial nitric oxide synthase
in human testis, epididymis, and vas deferens suggests a possi-
ble role for nitric oxide in spermatogenesis, sperm matura-
tion, and programmed cell death. Biology of Reproduction 55:
935–941.
Zweier JL, Samouilov A, Kuppusamy P (1999) Nonenzymatic
nitric oxide synthesis in biological systems. Biochimica et Bio-
physica Acta 1411: 250–262.
31