Asian Journal of Fisheries and Aquatic Research

Volume 26, Issue 6, Page 68-75, 2024; Article no.AJFAR.117446

ISSN: 2582-3760

Investigating the Acute Toxic Effects of

Urea Fertilizer on Juvenile Nile Tilapia
(Oreochromis niloticus)
George, U. U. a* and Essien-Ibok, M. A. b
a Department of Fisheries and Aquaculture, Faculty of Agriculture, Akwa Ibom State University, Obio
Akpa Campus, Akwa Ibom State, Nigeria.
b Department of Fisheries and Aquatic Environmental Management, University of Uyo, Nigeria.

Authors’ contributions

This work was carried out in collaboration between both authors. Both authors read and approved the
final manuscript.

Article Information
DOI: https://doi.org/10.9734/ajfar/2024/v26i6776

Open Peer Review History:

This journal follows the Advanced Open Peer Review policy. Identity of the Reviewers, Editor(s) and additional Reviewers,
peer review comments, different versions of the manuscript, comments of the editors, etc are available here:
https://www.sdiarticle5.com/review-history/117446

Received: 28/03/2024
Original Research Article Accepted: 31/05/2024
Published: 11/06/2024

ABSTRACT

The research aimed to assess the acute toxicity of urea fertilizer on juveniles of Oreochromis
niloticus. The findings indicated a correlation between increasing concentrations of urea fertilizer
and higher mortality rates among the test organisms, with an LC50 value determined at 500 mg/l.
Additionally, a rise in total dissolved solids was observed with higher concentrations of urea.
Parameters such as temperature, electrical conductivity, dissolved oxygen and pH showed
statistically significant (P < 0.05) changes as the concentrations of urea increases. Behavioural
responses during the exposure phase included heightened excitability, vigorous swimming
behaviours, loss of equilibrium, culminating in mortality. These results unequivocally demonstrate
that urea fertilizer at acute concentrations poses significant toxic risks to juvenile Oreochromis
niloticus.

_____________________________________________________________________________________________________

*Corresponding author: Email: [email protected];

Cite as: U. U., George, and Essien-Ibok, M. A. 2024. “Investigating the Acute Toxic Effects of Urea Fertilizer on Juvenile Nile
Tilapia (Oreochromis Niloticus)”. Asian Journal of Fisheries and Aquatic Research 26 (6):68-75.
https://doi.org/10.9734/ajfar/2024/v26i6776.
 George and Essien-Ibok; Asian J. Fish. Aqu. Res., vol. 26, no. 6, pp. 68-75, 2024; Article no.AJFAR.117446

Keywords: Acute toxicity; urea; juveniles; organic fertilizer; Oreochromis niloticus; agricultural
productivity.

1. INTRODUCTION organisms [9], while changes in water chemistry

and habitat structure can affect reproductive
The acute toxicity of fertilizers on aquatic success, growth rates and species diversity
organisms such as the Nile tilapia (Oreochromis [10,11].
niloticus) is a significant concern in aquatic
ecology and environmental management. The The relationship between fertilizers, water quality
use of fertilizers in agriculture has been and aquatic species is multifaceted and dynamic.
instrumental in boosting crop yields and meeting While fertilizers play a crucial role in agriculture
the demands of a growing population. However, production, their improper use can result in
the widespread application of fertilizers has detrimental effects on aquatic ecosystems.
raised concerns about its impacts on water Addressing this challenge requires a
quality and aquatic species. The intricate comprehensive approach, including improved
relationship between fertilizers, water quality and nutrients management practices, watershed
aquatic species revolves around the complex management strategies and conservation effort
dynamics of nutrients enrichment, ecosystem aimed at restoring and preserving the health of
responses and ecological consequences. aquatic environments.
Fertilizers primarily containing nitrogen and
phosphorous are essential for promoting plant O. niloticus is a widely cultured freshwater fish
growth and agricultural productivity. species known for its economic importance and
ecological role. By examining the impacts of
However, when these nutrients are applied fertilizers on Nile tilapia at acute exposure level,
excessively or mismanaged, they can infiltrate we aim to elucidate the potential risk posed by
water bodies via surface run-off, leaching or urea fertilizer contamination to aquatic
direct application in aquaculture ponds. As a ecosystems and highlight the importance of
results aquatic organisms are exposed to understanding and mitigating these effects for
elevated concentrations fertilizers derived sustainable aquatic resource management.
chemicals, which can have adverse effects on
their health behaviour and survival 2. MATERIALS AND METHODS
[1,2,3,4,5,6,7,8].
2.1 Source of Experimental Fish
Once in aquatic ecosystem, these nutrients fuel
the rapid growth of algae and other aquatic Juveniles of O. niloticus with mean weight (7.65
plants leading to a phenomenon known as ± 0.42g) were obtained from Myra fish farm
eutrophication. Algal blooms, fueled by nutrient- Gora, Nasarawa State. Nigeria. Fish were
rich run-off can blanket water surfaces, blocking transported in aerated plastic containers to the
sunlight and depleting oxygen levels as they hatchery of the Department of Fisheries and
decompose [6,7,8]. This oxygen depletion known Aquaculture, Joseph Sarwuan Tarka University,
as hypoxia or anoxia creates dead zones where Makurdi, Benue state. The fish were stocked in
aquatic life struggle to survive. 11 plastic containers of 45-liter capacity with well
aerated dechlorinated municipal tap water.
The impacts of eutrophication extend beyond
oxygen depletion as it is accompanied with 2.2 Acclimatization of Fish
harmful algal blooms, often dominated by toxin-
producing species pose risks to human health, Fish were acclimated to laboratory conditions for
aquatic organisms and entire ecosystems 7 days during which period they were fed twice
[5,6,7,8]. Additionally, the proliferation of algae daily (08.00am and 04.00pm) at five percent
can alter water chemistry, pH levels and light body weight with a commercial feed (vital feed).
penetration, further disrupting the balance of During this period, dead and abnormal fish were
aquatic ecosystem [3]. removed. The purpose of the acclimation was to
enable them recover from collection and
Aquatic species ranging from fish to transportation stresses and stabilized to the
invertebrates and aquatic plants face numerous experimental environment. Water was changed
challenges in eutrophic environments which may daily to discard faecal material and left-over food.
lead to massive mortality among fish and other Fish were not fed 24 hours prior to and during

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the exposure period. Mortality during the N = Total Individual Stocked

acclimation period was less than two percent. It
was from the acclimated population that healthy The difference between dead fish and
individuals used as test fish in this study were survivors will give the percentage survival of the
carefully selected. juveniles at the end of the experiment (96 hours)
[12].
2.3 Preparation of Test Media and
Exploratory Test 2.5.2 Determination of mortality lethal median
concentration (96 hours LC50)
To obtain the ranges of concentrations as used
in the experiment, five fishes were selected and The effects of the various concentrations of urea
each were exposed to 10 liters of dechlorinated fertilizer on the juveniles of O. niloticus was
tap water containing different weights of the determined by graphical method (Probit Level
fertilizer and used for the preliminary runs for Determination as recommended by Omoregie
twenty-four hours, until suitable concentration [15], Omoregie and Ufodike [16], Ekanem and
that resulted in 100% mortality was derived. The Ekpo [13] and Udo et.al. [12]. At Lethal Median
fish were not fed twenty-four hours before and Concentration LC50, after 96 hours of test, the
during these trials. The ranges of concentration number of fingerlings that are expected to die
values used in this study were determined from was determined from the graph. Similarly, the
the 100% mortality obtained from the trials. concentration that will kill 50% of the stocked
fingerlings at the end of the test (96 hours) was
2.4 Experimental Procedures determined at the probit level (Omoregie [15]
Omoregie and Ufodike [16], Udo et. al., [12],
The exposure concentrations of urea fertilizer Ekanem and Ekpo [13].
gotten from the trial include the following 400,
450, 500, 550 and 600 mg/L with dechlorinated 2.6 Water Quality Parameters
municipal water without fertilizer (0.00 mg/L)
which serves as the control. Ten fish were Water quality parameters that were measure
exposed to each of the five concentrations and include; Temperature, dissolved oxygen (DO),
each concentration had a replicate to ensure pH, electrical conductivity (EC) and total
accuracy. The exposure period lasted 96 hours. dissolved solid (TDS). Dissolve oxygen was
The effect of the acute concentrations of urea measured using DO meter model L933246 while
fertilizer on opercula ventilation, tail beat rate and other parameters were monitored every 24 hours
other behavioural characteristics were also using a multi parameter water quality tester
observed. Dead fish were removed immediately (Hanna meter, model Hi 98127), where the
to avoid fouling the test media and recorded probes were immersed directly into the water and
against the concentration. allowed to stabilize then readings were taken as
displayed on the screen.
2.5 Monitoring of Specimen for Mortality
2.7 Data Analysis
The effects of the various concentration of urea
fertilizer on the juveniles was monitored on a 24 Data collected were subjected to one way
hours’ basis for 96 hours as recommended by analysis of variance with Duncan’s new multiple
Udo et. al., [12] and Ekanem and Ekpo [13]. range post hoc-test for significance difference (P
< 0.05).
2.5.1 Determination of mortality and survival
rates of O. niloticus juveniles 3. RESULTS
The percentage mortality and survival rates of
the juveniles in the different concentrations of 3.1 Initial Water Quality Parameters
urea fertilizer during the period of study was
determine using the formula; The initial water quality parameters prior to
commencement of the toxicity studies are shown
% mortality =n/N × 100 [14]. in Table 1. The values of dissolved oxygen (4.9
mg/l), temperature (29.5 0C) and pH (6.80)
Where;
observed were within the acceptable range for
n = number of dead fish per aquarium per
aquaculture operations.
concentration

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Table 1. Initial physico-chemical parameters of the test water prior to stocking of test organism

Fish Species Initial physico-chemical parameters prior to stocking

DO (mg/l) Temp (oC) pH
Oreochromis niloticus 4.9 29.5 6.80

Table 2. Mean water quality parameters of the bioassay water for acute toxicity experiment
with urea fertilizer on Oreochromis niloticus juveniles

Concentration pH DO Temperature TDS EC

(mg/L)
0.00 8.10±0.10b 4.90±0.05a 28.15±0.05a 263.15±0.15d 527.20±0.20d
400.00 6.60±0.10d 3.90±0.10b 28.10±0.10a 664.05±0.05a 1434.20±0.25a
450.00 7.15±0.15c 3.40±0.10c 28.20±0.10a 629.15±0.15b 1266.20±0.15b
500.00 7.45±0.05c 3.20±0.10c 28.25±0.25a 446.15±0.15c 904.25±0.25c
550.00 8.80±0.00a 3.95±0.05b 27.40±0.10b
600.00 9.20±0.10a 4.80±0.10a 28.15±0.15a
P-Value 0.00 0.00 0.04 0.00 0.00
*Means in the same column with different superscripts differ significantly (P<0.05)

Table 3. Percentage mortality of Oreochromis niloticus juveniles exposed to acute

concentrations of urea fertilizer for 96hours

Concentration 6 Hrs. 12 Hrs. 24 Hrs. 72 Hrs. 96 Hrs. Total Mortality

(mg/L) Mortality (%) Mortality Mortality Mortality Mortality (%)
(%) (%) (%) (%)
0.00 0.00 0.00 0.00 0.00 0.00 0.00
400.00 0.00 10.00 0.00 10.00 0.00 20.00
450.00 10.00 0.00 10.00 10.00 0.00 30.00
500.00 20.00 20.00 10.00 0.00 0.00 50.00
550.00 30.00 20.00 10.00 20.00 0.00 80.00
600.00 30.00 30.00 20.00 10.00 0.00 90.00

Table 4. Mortality record of O. niloticus juveniles exposed to acute concentrations of urea

fertilizer for 96hours

Concentration Log10 Conc Total Number of Test Mortality Rate after 96 Percentage Mortality
(mg/l) Fish Hours (%)
0.00 0 10 0 0.00
400.00 2.60 10 2 20.00
450.00 2.65 10 3 30.00
500.00 2.70 10 5 50.00
550.00 2.74 10 8 80.00
600.00 2.78 10 9 90.00

3.2 Physico-chemical Parameters each of the concentrations are shown in Table 3

for the two batches of the experiment.
The results of the water physico-chemical
parameters of the experimental media (Table 2) At concentrations of 600.00 mg/l, 90% mortality
shows there was an increase in the total was recorded at the end of 96 hours of test
dissolved solid content as the urea (Table 3). At concentrations of 550.00, 500, 450
concentrations increased. There was no and 400 mg/l, the mortality recorded were 80, 50,
significant difference in temperature, there were 30 and 20% respectively at the end of 96 hours
however significant differences in the values of of test. No mortality was recorded in the control
electrical conductivity, dissolve oxygen and pH (0.00 mg/l) (Table 3) in both batches. There
(P < 0.05). was a positive correlation between fertilizer
concentration versus mortality which indicates
3.3 Mean Mortality of Juveniles Exposed that as concentration of fertilizer increased,
mortality rate also increased. Statistical Analysis
to Urea Fertilizer for 96 Hours
using one-way Anova (SPSS 20.0) showed that
there was no significant difference (p>0.05) in
The percentage mortality and survivors of O.
mortality between the two batches.
niloticus juveniles at the end of the test period in

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3.4 Determination of 96 Hours LC50 Results obtained from this research revealed that
the 96 h LC50 value of urea fertilizer for
The 96 hours LC50 for O. niloticus juveniles Oreochromis niloticus was 500.00 mg/L
exposed to the different concentrations of urea representing a log transform concentration of
fertilizer was determine using probit analysis. 2.74 mg/l. The result obtained was at variance
The concentrations were first transformed into with the 96 h LC50 of 1.78mg/l reported by
log for the probit analysis (Table 4). The 96 hours Essien-Ibok et.al., [18] when Heterobranchus
LC50 is given at 500.00 mg/l representing a log birdosalis was exposed to acute concentrations
transformed concentration of 2.74 mg/l a point of urea fertilizer. Also, the present findings do not
where 50 % of the test organisms would be killed align with LC50 reported by Asuquo, et. al., [19]
at the end of the experiment. when reporting on the effects of agricultural
fertilizers on fingerlings of Heterobranchus
3.5 General Behavioural Changes birdosalis. The difference may be due to
difference in fish species, levels of concentration
Behavioural changes occurred in the fish treated used and environmental conditions. Inorganic
with Urea fertilizer at different concentrations. fertilizers produce intermediate products that
The abnormal behaviours observed in fish may result in stress, fatigue, nervous disorder
exposed to concentrations of Urea fertilizers and death. This could be understandable as the
were characterized by restlessness, gulping of toxicity of chemicals depend on the type,
air, attempts to jump out of the tank and erratic composition, technical grade of preparations and
swimming before death. These behavioural the susceptibility of the exposed organisms [20].
changes showed by the exposed fish in response The results of the toxicity of urea fertilizer are
to the effect of the fertilizer was more indeed concentration dependent, meaning its
pronounced in tanks containing higher harmful effects increase as the concentration of
concentrations, but decreased with increase in urea increases. The results of the present study
time of exposure. There were no obvious agree favourably with the findings of previous
changes in fish behaviour in the lower authors [21,22,23,24,25,26,27,28] who reported
concentrations (400 mg/l) for the first 24 h of similar scenario of mortality in a concentration-
exposure. However, fish in the control group of dependent pattern.
the treatment did not exhibit any abnormal
behavior.
Water quality showed some variations during the
4. DISCUSSION, CONCLUSION AND test period. There were significant differences in
the mean values of some parameters measured
RECOMMENDATIONS
(P > 0.05). The air gulping and the attempt to
4.1 Discussion jump out of the tank observed in the exposed fish
is an indication of insufficient amount of
Urea is a widely used nitrogen fertilizer in dissolved oxygen in the experimental media. The
agriculture due to its high nitrogen content and variation observed in the water quality is as a
cost-effectiveness. Despite its agricultural result of the addition of urea fertilizer as no
benefits, urea can pose environmental and variation was observed in the control tank.
health risks, particularly when used Similar results have been documented by
inappropriately or in excessive amounts. In fish George [29,30,31,32,33,34,34,35].
farms, chemical fertilizers are often applied
before stocking the pond to stimulate the Behavioural responses of fish to most toxicants
production of organisms that may serve as first and differences in reaction times have been
food for many species of fish and also increase observed to be due to the effect of the chemical,
survival and growth [17]. Such applications may their concentrations, species, size and specific
not be harmful if enough time is allowed for the environmental conditions [21,22]. The
degradation of these fertilizers by the micro flora. behavioural responses reported for the test fish
In the context of fish nursery management, it in this study are similar to those reported by
would seem prudent to avoid situations where other authors for clarrids under various stress
chemical fertilizers are added intermittently to the conditions [36,37,38]. George et. al., [25,26]
ponds, because such subsequent additions may identified four main phases in the exposure time
result in total juveniles’ mortality, if the on behavioural responses of fish to toxicants.
concentrations exceed the established LC50 These are the contact phase (brief period of high
reported in this study. excitability), exertion (visible avoidance
characterized by fast swimming, leaping and

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attempts to jump out of the toxicant), loss of DISCLAIMER (ARTIFICIAL INTELLIGENCE)

equilibrium, followed by (death) phase, when
opercular movement and responses to tactile Author(s) hereby declare that NO generative AI
stimuli cease completely. In-spite of the technologies such as Large Language Models
numerous advantages of chemical fertilizers to (ChatGPT, COPILOT, etc) and text-to-image
improve fish production, they have a startling generators have been used during writing or
number of adverse effects on aquatic life in water editing of manuscripts.
bodies that receive run-off from farmlands or
from excess direct application in the aquatic COMPETING INTERESTS
environment [39].
Authors have declared that no competing
The stressful behaviours exhibited by the fish as
interests exist.
established in the study, suggest that they
suffered respiratory impairment, due to the effect
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