Length-weight relationship and condition factor of Nile tilapia with guava and star gooseberry leaf extract

F1000Research 2024, 13:540 Last updated: 24 DEC 2025
RESEARCH ARTICLE
Length-weight relationship and condition factor of Nile
tilapia (Oreochromis niloticus) fed diets supplemented with
guava and star gooseberry leaf extract
[version 2; peer review: 3 approved]
Manoj Tukaram Kamble 1,2, Krishna Rugmini Salin1,
Balasaheb Ramdas Chavan3, Seema Vijay Medhe2, Kim D. Thompson
Nopadon Pirarat 2
4,
1Aquaculture and Aquatic Resources Management (AARM), Department of Food, Agriculture and Bioresources, School of
Environment, Resources, and Development, Asian Institute of Technology, Khlong Nueng, Pathum Thani, 12120, Thailand
2Centre of Excellence in Wildlife, Exotic and Aquatic Animal Pathology, Faculty of Veterinary Science, Chulalongkorn University,
Bangkok, 10330, Thailand
3Department of Aquaculture, College of Fisheries, Ratnagiri, Maharashtra, 415629, India
4Moredun Research Institute, Pentlands Science Park, Penicuik, Scotland, EH26 0PZ, UK
v2 First published: 28 May 2024, 13:540
https://doi.org/10.12688/f1000research.145369.1
Latest published: 23 Oct 2024, 13:540
https://doi.org/10.12688/f1000research.145369.2
Open Peer Review
Approval Status
1
Abstract
Background
Nile tilapia (Oreochromis niloticus) is predominant cultured species in
aquaculture. However, there is a scarcity of literature regarding
relationship between guava and star gooseberry leaf extract and the
condition factor. Thus, the present study aims to investigate the effect
of guava and star gooseberry leaf extract-supplemented diets on the
specific growth rate, length-weight relationship, and condition factor
of Nile tilapia.
Methods
Six hundred and thirty Nile tilapia (8.7±0.4 g) were randomly
distributed among twenty-one tanks (30 fish per tank) within a
recirculation system. Over a 60-day period, the fish were fed diets
supplemented with 5g/Kg and 10g/Kg of guava leaf extract (GLE-5,
GLE-10), star gooseberry leaf extract (SGLE-5, SGLE-10), and a mixture
of both (MxLE-5, MXLE-10). Subsequently, specific growth rate, lengthweight relationship, and condition factor were determined.
2
3
version 2
(revision)
view
23 Oct 2024
version 1
28 May 2024
view
view
1. Dr.Sheikh Muhammad Azam, University of
Education Lahore, Punjab, Pakistan
2. Agumassie Tesfahun
, Debre Tabor
University, Debre Tabor, Ethiopia
3. Dr. Md. Abdullah-Al-Mamun, Sylhet
Agricultural University, Sylhet, Bangladesh
Any reports and responses or comments on the
article can be found at the end of the article.
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
Results
After 60 days, the specific growth rate was significantly higher in all
the GLE, SGLE, and MxLE groups compared to the control group. The
final lengths and weights differed significantly both in the control
group and all the GLE, SGLE, and MxLE groups. The analysis of the
regression equation indicated a positive correlation (r = 0.970, 0.977,
0.976, 0.974, 0.974, 0.974, and 0.969) between the length and weight
of Nile tilapia in the control group and in all the GLE, SGLE, and MxLE
groups. The regression exponent “b” values in all the GLE, SGLE, and
MxLE groups were >3, indicating a positive allometric growth pattern
in Nile tilapia compared to the control (b=2.866), which exhibited a
negative allometry. The final condition factor values did not differ
significantly in either the control group or any of the plant extract
groups.
Conclusions
Nile tilapia exhibited positive allometric growth patterns and
maintained good health when fed with GLE, SGLE, and MxLE groups.
Therefore, these plant extracts demonstrate suitability for commercial
Nile tilapia production.
Keywords
Nile tilapia, length-weight relationship, condition factor, specific
growth rate, guava, star gooseberry
This article is included in the Agriculture, Food
and Nutrition gateway.
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
Corresponding author: Nopadon Pirarat ([email protected])
Author roles: Kamble MT: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project
Administration, Resources, Software, Supervision, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review &
Editing; Salin KR: Conceptualization, Data Curation, Funding Acquisition, Methodology, Project Administration, Resources, Supervision,
Validation, Writing – Review & Editing; Chavan BR: Conceptualization, Funding Acquisition, Project Administration, Resources, Software,
Writing – Review & Editing; Medhe SV: Conceptualization, Formal Analysis, Investigation, Methodology, Software, Writing – Original Draft
Preparation, Writing – Review & Editing; Thompson KD: Conceptualization, Formal Analysis, Supervision, Visualization, Writing – Review
& Editing; Pirarat N: Conceptualization, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration,
Resources, Supervision, Writing – Review & Editing
Competing interests: No competing interests were disclosed.
Grant information: This research was funded by the Second Century Fund (C2F) Postdoctoral Fellowship, Chulalongkorn University,
Bangkok, Thailand and the Social Justice and Special Assistance Department, Government of Maharashtra, India.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Copyright: © 2024 Kamble MT et al. This is an open access article distributed under the terms of the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
How to cite this article: Kamble MT, Salin KR, Chavan BR et al. Length-weight relationship and condition factor of Nile tilapia (
Oreochromis niloticus) fed diets supplemented with guava and star gooseberry leaf extract [version 2; peer review: 3 approved]
F1000Research 2024, 13:540 https://doi.org/10.12688/f1000research.145369.2
First published: 28 May 2024, 13:540 https://doi.org/10.12688/f1000research.145369.1
Page 3 of 19
F1000Research 2024, 13:540 Last updated: 24 DEC 2025
REVISED Amendments from Version 1
We sincerely appreciate the reviewers’ insightful comments, which have greatly contributed to the improvement of the
second version of our manuscript. Based on their suggestions, we have made the following revisions:
First, we have included a Pearson correlation matrix to thoroughly analyze the relationship between water quality
parameters (e.g., temperature, dissolved oxygen, ammonia) and fish biometric parameters (length and weight). The results
of this analysis are now presented in a detailed tabular format.
Additionally, we have expanded the discussion section to provide a more comprehensive interpretation of the correlation
matrix, emphasizing the potential effects of water quality on fish growth and overall health. This enhanced analysis offers
deeper insights into the interrelationships between environmental conditions and fish biometrics, enriching the manuscript
and aligning it with the reviewers’ recommendations. We believe these revisions significantly strengthen the scientific rigor
and clarity of our study.
Any further responses from the reviewers can be found at the end of the article
Introduction
Nile tilapia (Oreochromis niloticus) is recognized as an important species in aquaculture within tropical and subtropical
regions, owing to its faster growth, heightened productivity, and notable resistance to diseases.1–4 In 2020, the global
production of Nile tilapia nearly reached 4.9 million metric tons (Mmt), and the continuous growth is attributed to the
robust worldwide demand.5 Furthermore, technological advancements with high stocking densities play a significant role
in the occurrence of disease outbreaks, resulting in substantial economic losses and impeding the sustainable progression
of tilapia culture globally.6
The incorporation of plant extracts into aquafeed is significant in finfish aquaculture7 because the indiscriminate use of
antibiotics has led to the development of drug-resistant bacteria.8,9 Plant extracts can serve as cost-effective and
environmentally sustainable prophylactic or antimicrobial agents. Guava (Psidium guajava) leaf (GLE) are rich in
various phenolic and flavonoid compounds10 and possess antimicrobial,11 antiviral,12 and antioxidant13 activities. GLE
have been employed as supplements in fish feed to boost the growth, immune response, and protection against diseases
for various fish species.14–20 Star gooseberry (Phyllanthus acidus) leaf (SGLE) contain flavonoids, kaempferol,
hypogallic acid, caffeic acid, and phenolic compounds.21,22 SGLE exhibited antioxidant, anti-inflammatory and
antimicrobial activities.23,24 It is also employed as a dietary supplement to augment the immune response of fish9,25
and chickens.26
The length-weight relationship (LWR) plays an essential role in estimating the biomass of diverse fish populations.27
Additionally, LWR is valuable for assessing fish condition and growth patterns.28 The condition factor (CF) contributes
to comprehending the development and adaptability of fish.29 A heightened condition factor indicates the robust health
and positive growth of the fish.30 Hence, the condition factor can be employed to assess the feeding activity of a species,
indicating its effective utilization of available feeding sources.27 Previous studies have investigated the length-weight
relationship and feeding habits of tilapia.27,30–33 Unfortunately, there is a scarcity of literature regarding relationship
between guava and star gooseberry leaf extract and the condition factor. Thus, the present study aims to investigate the
effect of guava and star gooseberry leaf extract-supplemented diets on the specific growth rate, length-weight relationship, and condition factor of Nile tilapia.
Methods
Chemicals
The analytical grade chemicals utilized for the determination of Ammonia, Nitrite, and Nitrate from the experimental
tanks included Sodium Hypochlorite 10% (Commercial, U&V Holding (Thailand) CO., LTD), Hydrochloric acid, 37%
(Fisher Scientific UK), Sodium hydroxide (Pine Chemical, Finland), Phenol, 99+% (Fisher Scientific UK), Sulfanilamide (Acros Organics Bvba, Belgium), N-(1-Naphthyl) ethylenediamine dihydrochloride (Alfa Aesar, USA), Cadmium
granular (ALDRICH Chemical, USA), Manganese (II) Sulphate H2O, Ammonium Chloride, Sodium Nitrite, Copper
(II) sulfate, and Ethylenediamine Tetra Acetic acid (Daejung Chemicals and Metals, South Korea), as well as Sodium
Tetraborate and Potassium Nitrate (KEMAUS, Australia) (Underlying Data).34
Preparation of guava and star gooseberry leaf aqueous extracts
The guava (P. guajava) and star gooseberry (P. acidus) leaf powder were received from All-Season Herbs Pvt. Ltd.,
Bangalore, India and the aqueous extract was prepared.35 Briefly, distilled water (100 mL) was mixed with leaf powder
(10 g) and homogenized using an orbital shaker (Hsiangtai D500) at 100 rpm for 20 h at room temperature (RT). The
resultant mixture underwent centrifugation at 8,000 rpm for 15 min at RT. A rotavapor (BÜCHI R-200/205) at 35 °C used
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to evaporate the supernatant and continue drying for 48 h in a hot air oven at 50 °C. The dried samples obtained were
placed in bottles and kept refrigerated at 4°C until further use.
Ethical statement
This study was conducted under the project entitled ‘Application of Phytobiotic Supplementation to Improve Disease
Resistance of Nile Tilapia (Oreochromis niloticus)’ and funded by the Social Justice and Special Assistance Department,
Government of Maharashtra, India, and Second Century Fund (C2F) Postdoctoral Fellowship, Chulalongkorn University, Bangkok, Thailand. This study was carried out in strict accordance with the recommendations of the Asian Institute
of Technology (AIT), Thailand, and the protocol was approved by the Local Ethical Committee for Experiments on
Animals of the AIT, Thailand (Project Number: AIT-AQ-18-01, May 2018–April 2019; April 29, 2018). Additionally,
according to Thailand Legislation, the Nile tilapia were not categorized as a protected species. The research was carried
out at the Laboratory of the Aquaculture and Aquatic Resources Management (AARM, Asian Institute of Technology
(AIT), Thailand, over 60 days from February to March, 2019. This study is reported in line with the ARRIVE guidelines.
A total of 630 fish were used in the experiment, with 90 fish allocated to each treatment. Each treatment comprised 30 fish
per replicate, totaling three replicates. Prior to sampling, the fish underwent a 12-hour fasting period to empty their
digestive tracts. In this study, to reduce stress on the experimental animals, the animals housed in the containers were
gently anesthetized using 1 mL of clove oil per 10 L of water for a period of 2–3 min, after which their weights were
measured. Following this, the fish were returned to their designated glass tanks according to their treatment and
replication groups.
Fish
Nile tilapia monosex fingerlings (8.7 0.4 g and 8.0 1.0 cm) were obtained from a GMP Certified fish farm, Ayutthaya
Province, Thailand. The research was conducted at the laboratory of Aquaculture and Aquatic Resources Management
(AARM), Asian Institute of Technology (AIT), Thailand. For a period of 15 days, the fish were acclimatized in three
fiberglass tanks (500 L) before the commencement of the experiment. Fish were fed pelleted feed twice daily at 4% of
their body weight. Following the acclimatization period, the fish were randomly distributed among twenty-one 150-L
glass tanks (30 fish per tank), all integrated into a recirculation system.
Preparation of experimental diets supplemented with guava and star gooseberry leaf aqueous extracts
Plant extracts (10 mg), dissolved in 1 mL distilled water, were mixed with commercial feed ((Charoen Pokphand (CP)7710), and spaghetti-like strands were prepared using a mincer (MITSUYAMA YC80B-4). Pellets approximately 5 mm
in length were formed and dried for 24 h at 50 °C in a hot air oven. The dried pellets were then stored at 4 °C until the end of
the experiment. The proximate composition of the commercial diet, including moisture, ash, crude lipid, and crude
protein, was 7.6%, 8.2%, 6.5%, and 30.5%, respectively.
Experimental design
In this study, a completely randomized design was employed, comprising seven experimental treatments: 5g/kg and
10g/kg of guava leaf extract (GLE-5, GLE-10), star gooseberry leaf extract (SGLE-5, SGLE-10), and a mixture of both
(MxLE-5, MXLE-10). The mixed diets (MxLE) were prepared with an equal proportion (1:1) of GLE and SGLE extracts.
The control group, on the other hand, was fed a diet without plant extract supplementation. Fish were fed twice daily at 4%
of their body weight. Prior to sampling, the fish underwent a 12-hour fasting period to empty their digestive tracts. In this
study, to reduce stress on the experimental animals, the animals housed in the containers were gently anesthetized using
1 mL of clove oil per 10 L of water for a period of 2–3 min, after which their weights were measured. Following this, the
fish were returned to their designated glass tanks according to their treatment and replication groups. The specific growth
rate, length-weight relationship, and condition factor were determined after 60 days of feeding.
Water quality parameters
During the experiment, a Eutech Cyberscan PC300 multi-parameter apparatus was employed to measure daily water
quality parameters, including temperature, pH, and dissolved oxygen (DO). The weekly concentrations of ammonianitrogen (NH3-N) were assessed using Phenate or indophenol method,36 nitrite-nitrogen (NO2-N) concentrations were
quantified through direct spectrophotometric assay,36 and nitrate-nitrogen (NO3-N) levels were determined employing
cadmium reduction method.36
Specific growth rate of Nile tilapia fed diets supplemented with GLE, SGLE and MxLE
The specific growth rate37 was evaluated after 60 days of feeding trials using the following equation
specific growth rate ðSGR,%=dayÞ ¼ ð100 ð ln Final weight ðgÞ – ln Initial weight ðgÞÞ=ðexperimental days ðTÞÞ
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Length-weight relationship (LWR) of Nile tilapia fed diets supplemented with GLE, SGLE and MxLE
Length-weight relationship (LWR)38 between the total length and body weight was estimated using the following
formula
W ¼ aLb
Where W is body weight of fish (g), L is the total length (cm), a is the exponent denotes the rate of weight change with
length (constant), and b is the slope representing the weight of one unit length. These values are estimated through the
linear regression equation, which is transformed by applying the natural logarithm (Log) to both sides.
Log W ¼ Log a þ b:Log L
Where b = 3, growth pattern is isometric. However, when b is greater or less than 3, an allometric pattern emerges. This
allometric pattern can be positive, indicating increase in length relative to body thickness, or negative, signifying an
increase in length relative to body thinness.28
Relative condition factor (Kn) of Nile tilapia fed diets supplemented with GLE, SGLE and MxLE
The health condition of the tilapia was assessed by evaluating relative condition factor (Kn) in all the treatments.
Kn ¼ Wo =Wc
where Wo = observed weight, and Wc = calculated weight.39 When Kn is greater than 1, it indicates favorable growth
conditions for the fish. Conversely, when Kn is less than 1, it suggests that the organism is in poorer growth condition
compared to an average individual of the same length.
Data analysis
The results were presented as means SE, and statistical analysis was conducted using IBM SPSS Statistics software
(SPSS, Inc., Version 29). The homogeneity of data was determined using Levine’s test. The impact of plant extract
treatments on specific growth rate was assessed through one-way analysis of variance (ANOVA), followed by Tukey
post-hoc tests for multiple comparisons. An independent t-test was used to determine the significant difference between
the initial and final length and weight of Nile tilapia with plant extract treatments. Linear regression analysis was
employed to evaluate the length-weight relationship. The Pearson correlation matrix was used to analyze relationship
between water quality and fish biometric parameters. A significance level of p < 0.05 was considered for statistical
significance.
Results
Water quality parameters
Water quality parameters such as Temperature, pH, DO, NH3N, NO2N, and NO3N, did not show significant difference
among the plant extract treatments and control during the experiment (Table 1). In all treatments, the average values for
temperature, pH, DO, NH3N, NO2N, and NO3N were 29.6 °C, 7.47, 5.49 mg/L, 0.15 mg/L, 0.13 mg/L, and 0.23 mg/L,
respectively. Raw data are available as underlying data (Tables 1-6).34
Table 1. Water quality parameters of Nile tilapia fed diets supplemented with plant extracts.
Control
GLE-5
GLE-10
SGLE-5
SGLE-10
MxLE-5
MxLE-10
Temp (°C)
30.60.15
30.70.16
30.80.16
30.70.17
30.80.16
30.80.16
30.50.16
pH
7.390.02
7.160.02
7.180.01
7.310.03
7.210.02
7.260.02
7.270.02
DO (mg/L)
4.690.05
4.150.05
4.010.09
4.570.15
4.450.06
4.060.06
4.540.05
NH3N (mg/L)
0.130.01
0.180.01
0.180.01
0.140.01
0.170.01
0.210.01
0.160.01
NO2N (mg/L)
0.070.01
0.090.01
0.110.01
0.130.01
0.120.01
0.080.01
0.090.01
NO3N (mg/L)
0.270.01
0.240.01
0.230.01
0.250.01
0.260.01
0.240.01
0.250.01
Temp: Temperature; DO: dissolved oxygen; NH3N: Ammonia-Nitrogen; NO2N: Nitrite-Nitrogen; NO3N: Nitrate-Nitrogen; GLE-5: 5g/kg of
guava leaf extract; GLE-10: 10g/kg of guava leaf extract; SGLE-5: 5g/kg of star gooseberry leaf extract; SGLE-10: 10g/kg of star gooseberry
leaf extract; MxLE-5: 5g/kg of mixture of GLE and SGLE; MxLE-10: 10g/kg of mixture of GLE and SGLE.
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Figure 1. Specific growth rate of Nile tilapia fed diets supplemented with plant extracts. Results with the means
of three replicates SE; Different superscripts in the bar indicate significant difference (p < 0.05).
Specific growth rate
The survival rate was higher than 95% in all treatments. The specific growth rate was significantly higher in all plant
extract treatments compared to the control group. GLE-10 exhibited a higher SGR, while MxLE-10 showed a lower SGR
in the plant extract treatments (Figure 1). Raw data are available as underlying data (Table 7).34
Length-weight relationship and growth pattern
The lengths were significantly different at the end of the experimental period in the control (t(148)=-30.327, p=0.01),
GLE-5 (t(148)=-37.193, p=0.01), GLE-10 (t(148)=-32.297, p=0.01), SGLE-5 (t(148)=-37.552, p=0.01), SGLE-10
(t(148)=-28.491, p=0.01), MxLE-5 (t(148)=-31.824, p=0.01), and MxLE-10 (t(148)=-38.139, p=0.01) groups (Table 2).
Table 2. Length-weight relationship, growth pattern, and condition factor of Nile tilapia fed diets
supplemented with plant extracts.
N
Control
GLE-5
GLE-10
SGLE-5
SGLE-10
MxLE-5
MxLE-10
75
75
75
75
75
75
75
LMin-Max (cm)
11.6-18.5
11.2-19.8
12.0-20.1
12.0-19.6
11.9-20.5
12.5-19.5
12.4-19.1
Wmin-max (g)
25.0-106.0
24.0145.0
28.0157.0
29.6132.0
28.8156.0
30.0134.0
31.0128.0
a
-1.612
-1.797
-1.880
-1.853
-1.758
-1.917
-1.815
b
2.866
3.023
3.101
3.067
3.003
3.135
3.036
SE (b)
0.085
0.077
0.081
0.083
0.082
0.085
0.091
CI (b)
2.6983.035
2.8693.177
2.9413.262
2.9013.232
2.8393.167
2.9653.304
2.8553.218
r
0.970
0.977
0.976
0.974
0.974
0.974
0.969
R2
0.940
0.955
0.953
0.949
0.948
0.949
0.939
p
0.001
0.001
0.001
0.001
0.001
0.001
0.001
t-test sig
0.001
0.001
0.001
0.001
0.001
0.001
0.001
Growth
behavior
Negative
allometry
Positive
allometry
Positive
allometry
Positive
allometry
Positive
allometry
Positive
allometry
Positive
allometry
Kn
1.004
1.002
1.003
1.003
1.004
1.003
1.003
Min-Max
0.8651.177
0.8861.178
0.8241.097
0.8401.282
0.8171.162
0.8571.136
0.8221.162
SE
0.010
0.008
0.008
0.010
0.011
0.009
0.009
N: sample size; L: length (cm); W: weight (g); Min: minimum; Max: maximum; a: intercept; b: slope of the equation; SE: standard error; CI (b):
confidence intervals of b; r2: coefficient of determination; p: significance of the regression, with p significant at <0.05; Kn: relative condition
factors. Significance was determined by the t-test to verify if b was significantly different from the consensus (b = 3). The growth behavior
was based on b. GLE-5: 5g/kg of guava leaf extract; GLE-10: 10g/kg of guava leaf extract; SGLE-5: 5g/kg of star gooseberry leaf extract;
SGLE-10: 10g/kg of star gooseberry leaf extract; MxLE-5: 5g/kg of mixture of GLE and SGLE; MxLE-10: 10g/kg of mixture of GLE and SGLE.
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Figure 2. Final logarithmic length-weight relationship with regression equation for Nile tilapia fed plant
extracts.
Additionally, the weights were significantly different at the end of the experimental period in the control (t(148)=-31.206,
p=0.01), GLE-5 (t(148)=-38.611, p=0.01), GLE-10 (t(148)=-32.694, p=0.01), SGLE-5 (t(148)=-37.627, p=0.01),
SGLE-10 (t(148)=-30.032, p=0.01), MxLE-5 (t(148)=-32.800, p=0.01), and MxLE-10 (t(148)=-38.197, p=0.01) groups.
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Table 3. Correlation matrix between water quality and fish biometric parameters.
Parameter
Length
Weight
**
Temperature
DO
NH3-N
pH
NO2-N
NO3-N
*
-0.323
Length
1
0.969
0.364
-0.199
-0.312
0.403
0.489
Weight
-
1
0.339
-0.215
-0.354
0.470*
0.489*
-0.258
**
**
Temperature
-
-
1
-0.684
-0.060
0.426
-0.322
DO
-
-
-
1
-0.563
0.887**
-0.185
-0.269
0.331
pH
-
-
-
-
1
-0.328
-0.385
0.326
NH3-N
-
-
-
-
-
1
-0.189
0.071
NO2-N
-
-
-
-
-
-
1
-0.207
NO3-N
-
-
-
-
-
-
-
1
**Correlation is significant at the 0.01 level (2-tailed).
*Correlation is significant at the 0.05 level (2-tailed).
The analysis of the regression equation indicated a positive correlation (r = 0.970, 0.977, 0.976, 0.974, 0.974, 0.974, and
0.969) between the length and weight of Nile tilapia fed control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, MxLE10 diets, respectively.
The regression exponent of “b” values in GLE-5 (b=3.023, t(73)=39.216, p=0.01), GLE-10 (b=3.101, t(73)=38.515,
p=0.01), SGLE-5 (b=3.067, t(73)=36.981, p=0.01), SGLE-10 (b=3.003, t(73)=36.451, p=0.01), MxLE-5 (b=3.135, t(73)
=36.816, p=0.01), and MxLE-10 (b=3.036, t(73)=33.390, p=0.01) were >3, indicating a positive allometric growth
pattern in Nile tilapia compared to the control (b=2.866, t(73)=33.881, p=0.01), which exhibited negative allometry. The
length-weight correlation regression graphs for the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, MxLE-10
diets are depicted in Figure 2. Raw data are available as underlying data (Tables 8-14).34
Condition factor
The condition factors were not significantly different at the end of the experimental period in the control (t(148)=-0.599,
p=0.275), GLE-5 (t(148)=-0.199, p=0.421), GLE-10 (t(148)=0.299, p=0.383), SGLE-5 (t(148)=0.818, p=0.207),
SGLE-10 (t(148)=0.173, p=0.431), MxLE-5 (t(148)=0.371, p=0.356), and MxLE-10 (t(148)=-0.100, p=0.460) groups
(Table 2). Additionally, the minimum and the maximum values of condition factor for control, GLE-5, GLE-10, SGLE-5,
SGLE-10, MxLE-5, and MxLE-10 groups were 0.865-1.177, 0.886-1.178, 0.824-1.097, 0.840-1.282, 0.817-1.162,
0.857-1.136, and 0.822-1.162, respectively. Raw data are available as underlying data (Tables 8-14).34
Correlation matrix for water quality and fish biometric parameters
The results of Pearson’s correlation analysis are presented in Table 3. A moderate positive correlation was observed
between fish length and both ammonia (r = 0.403) and nitrite (r = 0.489), with the nitrite correlation being statistically
significant at the 0.05 level (Table 3). This finding suggests that as fish increase in length, there may be a corresponding
rise in ammonia and nitrite levels, potentially due to metabolic byproducts. Similarly, fish weight exhibited a positive
correlation with ammonia (r = 0.470) and nitrite (r = 0.489), with the latter correlation also significant at the 0.05 level.
These results indicate that fish biomass may contribute to the elevation of these water quality parameters.
Furthermore, a significant negative correlation (r = -0.684) was identified between temperature and dissolved oxygen at
the 0.01 level, indicating that higher temperatures are associated with reduced dissolved oxygen levels. This phenomenon
is well-documented, as warmer water typically holds less oxygen. Additionally, temperature demonstrated a moderate
negative correlation with pH (r = -0.563), which was also significant at the 0.01 level. Elevated temperatures may
influence the ionization of water, leading to decreased pH levels.
A strong positive correlation (r = 0.887) was observed between dissolved oxygen and pH, significant at the 0.01 level.
Higher dissolved oxygen levels are often associated with elevated pH in aquatic systems, likely due to increased
photosynthetic activity.
Discussion
Aquafeed supplementation with herbal or plant extracts has proven to be an effective means of enhancing the growth
performance, feed utilization, and immune response in farmed fish species.40,41 The supplementation of natural bioactive
rich-plant extracts may boost natural immunity and improve disease resistance,40 resulting in higher production and
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
profitability in the aqua industry. The length-weight relationship is used to predict the weight of fish in proportion to their
length over a certain period.38 Therefore, the present study investigated the effects of plant extract-supplemented diets on
the length-weight relationship and condition factor of Nile tilapia.
The water quality parameters in all treatments were within the acceptable range, consistent with the findings of previous
studies on spotted snakehead (Channa punctata) under various feeding regimes involving agro-industrial by-products,42
and on silver barb (Barbonymus gonionotus) fingerlings fed with dietary blanched moringa leaf meal.43 The specific
growth rate showed a significant increase in all plant extract treatments. Similarly, the specific growth rate of spotted
snakehead (C. punctata) and silver barb (B. gonionotus) was considerably improved under various feeding regimes
involving agro-industrial by-products42 and dietary blanched moringa leaf meal.43
Aqua farmers and consumers prefer fish with isometric growth or body structure.27,44 Furthermore, the length-weight
relationship provides insights into the growth and health of a fish species.45,46 In the present study, plant extractsupplemented diets observed significant correlation between length and weight of Nile tilapia. Our results are corroborated with previous study involving different maltose levels fed to Nile tilapia (O. niloticus).27 Importantly, Nile tilapia
fed diets supplemented with GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and MxLE-10 groups exhibited positive
allometric growth pattern compared to the control. Similarly, silver barb (B. gonionotus) and zebrafish (Danio rerio) fed
with dietary blanched moringa leaf meal43 and plant-protein based diets47 found positive allometric growth. On the
contrary, gold fish (Carassius auratus) fed with different concentrations of carrot48 and gurami sago (Osphronemus
goramy) stocked in concrete ponds, floating net cages, and earthen freshwater ponds,49 found negative allometric growth.
In the present study, plant extracts diets revealed a significant improvement in specific growth rate and positive allometric
growth patterns of Nile tilapia. This can be ascribed to the presence of bioactive compounds in GLE and SGLE, including
gallic acid, p-coumaric acid, quercetin, and kaempferol. These compounds have been demonstrated to enhance the
growth of various fish species, including common carp Cyprinus carpio,50 snakehead fish Channa argus,51 grass carp
Ctenopharyngodon idellus,52 as well as certain animals.53,54 Further research should be conducted in pond or cage culture
systems to substantiate the positive allometric growth patterns exhibited by Nile tilapia when fed diets supplemented with
GLE and SGLE.
The mean Kn values for both the plant extract diets and control were higher than one, indicating that the Nile tilapias were
in good health condition during the experimental period. Similarly, condition factor values were more than one when
Sargassum meal fed to Nile tilapia O. niloticus,31 Mansoa alliacea hydroalcoholic extracts fed to pirarucu Arapaima
gigas,55 different maltose levels fed to Nile tilapia O. niloticus.27
The strong correlations between fish length, weight, and specific water quality parameters, particularly ammonia and
nitrite, suggest a potential interplay between fish growth and water quality dynamics. As fish grow, their metabolic waste
production, including ammonia, increases, which can accumulate in the environment if not adequately managed. This
accumulation of ammonia and nitrite can adversely affect water quality and fish health, ultimately impacting growth rates
and overall well-being. Several studies support these observations. Elevated ammonia concentrations are known to
impair fish growth by disrupting metabolic processes and diminishing feeding efficiency. For instance, in the rockfish
Sebastes schlegelii, exposure to high ammonia concentrations resulted in reduced growth performance.56 Similarly,
increased nitrite levels have been associated with oxidative stress and compromised respiratory function in fish, which
can further inhibit growth.57 In aquaculture systems, higher fish densities or insufficient water treatment often exacerbate
the accumulation of ammonia and nitrite. Such conditions have been shown to decrease feed consumption and utilization
efficiency, as demonstrated in studies examining the effects of stocking density on water quality and fish performance.57,58 These correlations underscore the necessity for meticulous management of water quality parameters in
aquaculture to ensure optimal growth and health outcomes for fish populations.
Conclusion
The specific growth rate was significantly higher in GLE, SGLE, and MxLE groups compared to the control group. The
analysis of the regression equation indicated a positive correlation between the length and weight of Nile tilapia in the
control group and the GLE, SGLE, and MxLE groups. Furthermore, the GLE, SGLE, and MxLE groups exhibited a
positive allometric growth pattern in Nile tilapia compared to the control. The final condition factor values did not differ
significantly between the control group and any of the GLE, SGLE, and MxLE groups. Consequently, these plant extracts
demonstrate suitability for commercial Nile tilapia production.
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
Data availability
Underlying data
zenodo: Length-weight relationship and condition factor of Nile tilapia (Oreochromis niloticus) fed diets supplemented
with guava and star gooseberry leaf extract. https://doi.org/10.5281/zenodo.11174309.34
This project contains the following underlying data:
- Table 1: Weekly average temperature (°C) of the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and
MxLE-10 groups.
- Table 2: Weekly average pH of the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and MxLE-10
groups.
- Table 3: Weekly average DO (mg/L) of the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and MxLE10 groups.
- Table 4: Weekly average NH3N (mg/L) of the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and
MxLE-10 groups.
- Table 5: Weekly average NO2N (mg/L) of the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and
MxLE-10 groups.
- Table 6: Weekly average NO3N (mg/L) of the control, GLE-5, GLE-10, SGLE-5, SGLE-10, MxLE-5, and
MxLE-10 groups.
- Table 7: Specific growth rate of Nile tilapia (N=75) fed diets supplemented with control, GLE-5, GLE-10,
SGLE-5, SGLE-10, MxLE-5, and MxLE-10 for 60 days.
- Table 8: Initial and final length (cm), weight (g), and condition factor of Nile tilapia (N=75) fed diet
supplemented with control for 60 days.
- Table 9: Initial and final length (cm), weight (g), condition factor of Nile tilapia (N=75) fed diet supplemented
with GLE-5 for 60 days.
- Table 10: Initial and final length (cm), weight (g), and condition factor of Nile tilapia (N=75) fed diet
supplemented with GLE-10 for 60 days.
- Table 11: Initial and final length (cm), weight (g), and condition factor of Nile tilapia (N=75) fed diet
supplemented with SGLE-5 for 60 days.
- Table 12: Initial and final length (cm), weight (g), and condition factor of Nile tilapia (N=75) fed diet
supplemented with SGLE-10 for 60 days.
- Table 13: Initial and final length (cm), weight (g), and condition factor of Nile tilapia (N=75) fed diet
supplemented with MxLE-5 for 60 days.
- Table 14: Initial and final length (cm), weight (g), and condition factor of Nile tilapia (N=75) fed diet
supplemented with MxLE-10 for 60 days.
- Table 15: Summary of the chemicals and reagents used in the experiment.
- Authors checklist Manuscript No. 145369 (completed ARRIVE checklist).
Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).
Page 11 of 19
F1000Research 2024, 13:540 Last updated: 24 DEC 2025
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Open Peer Review
Current Peer Review Status:
Version 2
Reviewer Report 30 January 2025
https://doi.org/10.5256/f1000research.173657.r362059
© 2025 Abdullah-Al-Mamun D. This is an open access peer review report distributed under the terms of the
Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any
medium, provided the original work is properly cited.
Dr. Md. Abdullah-Al-Mamun
Professor, Fish Health Management, Sylhet Agricultural University, Sylhet, Sylhet Division,
Bangladesh
The manuscript is well-written, with a clear research objective and structured methodology. The
study provides valuable insights into the effects of guava and star gooseberry leaf extract on the
growth performance and condition factor of Nile tilapia. The findings, particularly the positive
allometric growth patterns and improved specific growth rate, contribute to the field of
aquaculture nutrition. However, the authors could extend their research by evaluating disease
resistance to determine whether these plant extracts offer any significant benefits in enhancing
the immune response of Nile tilapia. My recommendation is for approval, with this suggestion for
future studies.
Is the work clearly and accurately presented and does it cite the current literature?
Yes
Is the study design appropriate and is the work technically sound?
Yes
Are sufficient details of methods and analysis provided to allow replication by others?
Yes
If applicable, is the statistical analysis and its interpretation appropriate?
I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?
Yes
Are the conclusions drawn adequately supported by the results?
Yes
Page 14 of 19
F1000Research 2024, 13:540 Last updated: 24 DEC 2025
Competing Interests: No competing interests were disclosed.
Reviewer Expertise: I specialize in aquaculture, finfish vaccination, aquaculture nutrition, fish
immunology, and pathology. My research focuses on improving fish health and disease resistance
through nutritional and immunological interventions.
I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard.
Version 1
Reviewer Report 06 September 2024
https://doi.org/10.5256/f1000research.159308.r320437
© 2024 Tesfahun A. This is an open access peer review report distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited.
Agumassie Tesfahun
Debre Tabor University, Debre Tabor, Ethiopia
I advice the authors to include correlation matrix or principal component analysis (PCA) regarding
the water quality parameter and fish biometric relationships (length and weight). To this end,
please refer other published works in the same title. The paper is sound, however, it should be
more sound if the authors include more statistical analysis such as correlation matrix or principal
component analysis (PCA) to know the relationship between environmental variables and fish
biometric data. Except for this issue, the paper is well presented and innovative for the scientific
community.
Capture fishery is now declining due to human induced impacts on the water bodies. Aquaculture
is the alternative option to feed this fast-growing population. Therefore, this work is very
important for the fish farmers and the community as well for food security.
Is the work clearly and accurately presented and does it cite the current literature?
Yes
Is the study design appropriate and is the work technically sound?
Yes
Are sufficient details of methods and analysis provided to allow replication by others?
Yes
If applicable, is the statistical analysis and its interpretation appropriate?
Partly
Page 15 of 19
F1000Research 2024, 13:540 Last updated: 24 DEC 2025
Are all the source data underlying the results available to ensure full reproducibility?
Yes
Are the conclusions drawn adequately supported by the results?
Yes
Competing Interests: No competing interests were disclosed.
Reviewer Expertise: Assistant Professor in Fisheries and Aquatic Sciences
I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard.
Author Response 17 Oct 2024
nopadon pirarat
Comment 1: I advice the authors to include correlation matrix or principal component
analysis (PCA) regarding the water quality parameter and fish biometric relationships
(length and weight). To this end, please refer other published works in the same title. The
paper is sound, however, it should be more sound if the authors include more statistical
analysis such as correlation matrix or principal component analysis (PCA) to know the
relationship between environmental variables and fish biometric data. Except for this issue,
the paper is well presented and innovative for the scientific community.
Capture fishery is now declining due to human induced impacts on the water bodies.
Aquaculture is the alternative option to feed this fast-growing population. Therefore, this
work is very important for the fish farmers and the community as well for food security.
Response: Thank you for your valuable feedback and appreciation of our work. We have
incorporated a correlation matrix in the revised manuscript to analyze the relationships
between water quality parameters and fish biometric data, specifically length and weight.
This addition strengthens our analysis and aligns with your suggestion for more robust
statistical evaluation.
Your point regarding the declining capture fishery due to human-induced impacts is well
taken. As aquaculture emerges as a vital alternative for sustaining the growing global
population, our research aims to provide essential insights for fish farmers and contribute
to food security efforts within communities.
We appreciate your recognition of the significance of this work and look forward to any
further suggestions you may have.
In the revised manuscript
Correlation matrix for water quality and fish biometric parameters
The results of Pearson’s correlation analysis are presented in Table 3. A moderate positive
correlation was observed between fish length and both ammonia (r = 0.403) and nitrite (r =
0.489), with the nitrite correlation being statistically significant at the 0.05 level (Table 3).
This finding suggests that as fish increase in length, there may be a corresponding rise in
ammonia and nitrite levels, potentially due to metabolic byproducts. Similarly, fish weight
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
exhibited a positive correlation with ammonia (r = 0.470) and nitrite (r = 0.489), with the
latter correlation also significant at the 0.05 level. These results indicate that fish biomass
may contribute to the elevation of these water quality parameters.
Furthermore, a significant negative correlation (r = -0.684) was identified between
temperature and dissolved oxygen at the 0.01 level, indicating that higher temperatures are
associated with reduced dissolved oxygen levels. This phenomenon is well-documented, as
warmer water typically holds less oxygen. Additionally, temperature demonstrated a
moderate negative correlation with pH (r = -0.563), which was also significant at the 0.01
level. Elevated temperatures may influence the ionization of water, leading to decreased pH
levels.
A strong positive correlation (r = 0.887) was observed between dissolved oxygen and pH,
significant at the 0.01 level. Higher dissolved oxygen levels are often associated with
elevated pH in aquatic systems, likely due to increased photosynthetic activity.
Discussion
The strong correlations between fish length, weight, and specific water quality parameters,
particularly ammonia and nitrite, suggest a potential interplay between fish growth and
water quality dynamics. As fish grow, their metabolic waste production, including ammonia,
increases, which can accumulate in the environment if not adequately managed. This
accumulation of ammonia and nitrite can adversely affect water quality and fish health,
ultimately impacting growth rates and overall well-being. Several studies support these
observations. Elevated ammonia concentrations are known to impair fish growth by
disrupting metabolic processes and diminishing feeding efficiency. For instance, in the
rockfish Sebastes schlegelii, exposure to high ammonia concentrations resulted in reduced
growth performance. 56 Similarly, increased nitrite levels have been associated with
oxidative stress and compromised respiratory function in fish, which can further inhibit
growth. 57 In aquaculture systems, higher fish densities or insufficient water treatment
often exacerbate the accumulation of ammonia and nitrite. Such conditions have been
shown to decrease feed consumption and utilization efficiency, as demonstrated in studies
examining the effects of stocking density on water quality and fish performance. 57, 58
These correlations underscore the necessity for meticulous management of water quality
parameters in aquaculture to ensure optimal growth and health outcomes for fish
populations.
References
56) Shin KW, Kim SH, Kim JH, Hwang SD, Kang JC: Toxic effects of ammonia exposure on
growth performance, hematological parameters, and plasma components in rockfish,
Sebastes schlegelii, during thermal stress. Fish Aquat Sci. 2016;19:1–8. 10.1186/s41240-0160044-6.
57) Al-Harbi AH, Siddiqui AQ: Effects of tilapia stocking densities on fish growth and water
quality in tanks. Asian Fish Sci. 2000;13(4):391–396. 10.33997/j.afs.2000.13.4.011.
58) Sanders E, Farmer SC: Aquatic models: Water quality and stability and other
environmental factors. ILAR J. 2019;60(2):141–149. 10.1093/ilar/ilaa008.
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
Competing Interests: No competing interests were disclosed.
Reviewer Report 07 August 2024
https://doi.org/10.5256/f1000research.159308.r303303
© 2024 Azam D. This is an open access peer review report distributed under the terms of the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the
original work is properly cited.
Dr.Sheikh Muhammad Azam
Department of Zoology, Division of Science and Technology, Lahore, University of Education
Lahore, Punjab, Pakistan
Work will attract the scientist in the field. The results will be helpful for farmers and aquaculturists.
Is the work clearly and accurately presented and does it cite the current literature?
Yes
Is the study design appropriate and is the work technically sound?
Yes
Are sufficient details of methods and analysis provided to allow replication by others?
Yes
If applicable, is the statistical analysis and its interpretation appropriate?
Yes
Are all the source data underlying the results available to ensure full reproducibility?
No
Are the conclusions drawn adequately supported by the results?
Partly
Competing Interests: No competing interests were disclosed.
Reviewer Expertise: fisheries, wildlife
I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard.
Author Response 17 Oct 2024
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F1000Research 2024, 13:540 Last updated: 24 DEC 2025
nopadon pirarat
Comment 1: Work will attract the scientist in the field. The results will be helpful for farmers
and aquaculturists.
Response: Thank you for taking the time to review our manuscript. We appreciate your
positive feedback and are glad to hear that you believe our work will engage scientists in
the field. We aim to provide valuable insights that will benefit both farmers and
aquaculturists, contributing to more sustainable practices in aquaculture. Your
encouragement reinforces the importance of our research, and we look forward to sharing
our findings with the broader community.
Competing Interests: No competing interests were disclosed.
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