1.
INTRODUCTION
The economic value of algae has gained attention due to
their diverse applications in various industries, including aquaculture. In
fish feed, algae offer numerous benefits: they provide high-quality protein and
essential amino acids, improve feed conversion rates, and contain bioactive
compounds with immunomodulatory and anti-inflammatory properties (Veluchamy and Palaniswamy, 2020). Algae are an excellent
source of omega-3 fatty acids, which improve nutrition and flesh quality in fish.
By adding algae to fish diets, sustainable aquaculture practices can be
supported, as this reduces the need for fishmeal and helps lessen the pressure
on wild fish populations. In summary, including algae in aquaculture is
beneficial as it enhances fish health and promotes sustainability (Liu et al.,
2023; Rathod et al., 2025). Algae, as primary producers, are
essential to global aquatic ecosystems; they sustain marine food webs and
significantly contribute to carbon dioxide fixation and oxygen production (Pereira, L. 2021; Vijayaram et al., 2024).
They are utilized for sustenance, pharmaceuticals, cosmetics, and biofuels,
among other applications. The seashore of Pakistan is home to around 70 genera and
27 distinct kinds of algae, predominantly seaweeds (Siddiqui et al., 2019).
Based on color, algae fall into three main groups:
brown algae (Phaeophyta), green algae (Chlorophyta), and red algae (Rhodophyta)
(Shahzad et al., 2023). Among the most important economic algae in Phaeophyta
is sargassum. It provides a wide range of development and application prospects
in industry, agriculture, food, medicine, and ecological restoration (Zou et
al., 2022), so helping greatly to preserve the marine ecology. There are
more than 400 different species of brown seaweed in the genus Sargassum, and
each has special ecological use. Common in tropical locations, it produces
steroids, polysaccharides, and polyphenols. Bioactive polysaccharides derived
from brown seaweeds have gained significant interest among pharmacology and
biochemistry researchers. Brown seaweed-derived sulfated polysaccharides have
shown anti-inflammatory, antiviral, antioxidant, anti-cancer, and anticoagulant
properties (Jyotsna et al., 2021).
Aquaculture,
which has been utilized for centuries to produce food, is currently expanding rapidly
(Eroldogan et al., 2022). Fisheries provide food for humans and
industrial raw materials for national construction, contributing significantly
to the national economy and contributing to the country's economy (Hu et
al., 2021). Indian major Carp (Labeo rohita), commonly
known as rohu, is profitable from an economic standpoint, other Labeo species
could also be cultivated because of their excellent nutritional content,
mouthwatering flavor, low economic worth, and high market value. L. rohita
is one of the fish species that is most often raised in Asia (Khizar et al.,
2024).
Sargassum is a desirable addition to the fish diet since it is a bioactive
component that is rich in vitamins, minerals, and amino acids (Morais et
al., 2020). It stimulates immune responses, fortifies antioxidant defences,
and improves growth performance. Sargassum improves fish health and resilience
to environmental stressors by increasing nutritional absorption, disease
resistance, stress tolerance, and digestive enzyme activity. It also improves
meat quality and lipid metabolism, rendering fish suitable for human
consumption (Saade et al., 2020; Sabzi et
al., 2023; Gazali et al., 2024).
This study aims to examine the impact of S. ilicifolium on the
hematological parameters, growth performance, and immune response of L.
rohita. Standard fish feed ingredients, which often rely on expensive and
scarce resources such as fishmeal and soy protein, may be partially replaced with
the sustainable and cost-effective feed additive Sargassum. Given the abundance
of sargassum in tropical and subtropical regions, its application in
aquaculture reduces dependence on imported feed sources, thus decreasing
production costs for fish farmers.
2.
MATERIALS AND METHODS
This experimental study evaluated the effect of S.
ilicifolium supplementation on the growth performance and hematology of L.
rohita. It was carried out at the Fisheries Laboratory, Department of
Zoology, Government College University Faisalabad, for 90 days.
Collection of S.
Ilicifolium
Brown
seaweed (S. ilicifolium) was collected from Manora Beach, Karachi. The
seaweed was thoroughly washed under tap water to remove any debris or
impurities, followed by another wash with distilled water to ensure complete
cleanliness. The cleaned seaweed was dried and ground into a fine powder. This
powdered seaweed material was stored at room temperature until it was ready for
further analysis.
Experiment
Design
One control and three experimental diets were
prepared with varying amounts of sargassum powder in them. Three hundred fingerlings (2.36g) of L. rohita were
purchased from Punjab Fish Hatchery, Satyana Road, Faisalabad. Fish were
acclimatized for two weeks in the Fish Lab. During acclimatization, fish were
fed the control diet of 4% of their body weight once a day. After 15 days, fish
were randomly stocked in four aquariums: Control (0%BSW), T1(3%BSW), T2(5%BSW),
and T3 (7%BSW), each with triplicates (25 fingerlings per aquarium), arranged
using interspersion. Fish were fed with 4 % of their live body weights once a
day. Feeding behaviour was also observed in the trial.
Study of Growth
Parameters
Total
body length and weight were measured fortnightly over 90 days to track growth
performance metrics, according to the approach of Al Sulivany et al. (2024). After
data collection, fish were stocked back into respective aquariums. The ethics committee
of the Government College University Faisalabad, Pakistan, approved the animal
study protocol for scientific purposes. Approval no. is GCUF/ERC/505.
The following
formulas were used to calculate the growth parameters;
Analysis of
Hematology
Blood
samples were collected from the caudal vein of fish to assess the blood
indices. The total number of red blood cells (RBCs) and white blood cells
(WBCs) were assessed using a hemocytometer with a validated Neubauer counting
chamber (Blaxhall & Daisley, 1973). Hemoglobin concentration estimates were
obtained using the cyanomethemoglobin method, as outlined by Wedemeyer &
Yastuke (1977). The microhematocrit technique (Brown et al.,
1980) was employed to ascertain hematocrit using capillary tubes. The
percentage composition of leukocytes was determined based on the identification
characters listed (Ivanova, 1983). The subsequent factors were employed for
calculation: MCHC (mean corpuscular hemoglobin concentration), MCH (mean
corpuscular hemoglobin), and MCV (mean cell volume) utilizing the specified
formulas.
a.
Statistical Analysis
After
the possible outcomes were known, data on growth and hematology were subjected
to statistical analysis. ANOVA and ± SE values were computed by using GraphPad ver.
6.0, and the difference between means was calculated by Tukey’s HSD test.
3.
RESULTS
Growth Parameters
The
effects of S. ilicifolium on the growth performance of L. rohita
are shown in Table 1. After the 90-day trial, Weight gain (WG) and Specific
growth rated (SGR) were significantly higher (P<0.05) in groups fed with 5%(T2)
and 7% (T3) of S. ilicifolium compared to the control, with the highest
WG of 2.96 g and SGR of 0.90 observed in the T3. In contrast, the control group
exhibited a minimum WG of 1.57 g. The highest Protein Efficiency Ratio (PER)
value of 0.092 was recorded in the 7% fed group, significantly higher compared
to the control group’s PER of 0.49. However, the highest condition factor was
observed in the control group, and the lowest was observed in the T3 treatment.
Overall, these results suggest the beneficial effects of S. ilicifolium
on the growth performance of L. rohita at higher dietary levels.
4.
DISCUSSION
In
the present growth study, the inclusion of Sargassum spp. in the feed presented
a significant improvement in growth rates associated with the control set.
Among the different inclusion levels, the group with the highest inclusion (7%)
showed the best growth performance, with the maximum final body weight (FBW),
weight gain (WG), and specific growth rate (SGR). The present findings are similar
to the studies in which tilapia showed better growth, WG and SGR when fed with
20 g/kg and 30 g/kg sargassum than control group (Abdelrhman et al.,
2022), similarly Shi et al. (2021) stated that yellow catfish
when fed with Sargassum hornei extract in feed enhanced growth and
significant improvement in specific growth rate (SGR) and weight gain (WG) was
observed. Yeganeh and Adel (2019) observed that average body weight, total body
length and SGR enhance when supplemented with S. ilicifolium extract
particularly at 7.5% inclusion. These results show a favorable association
between increased seaweed inclusion and enhanced fish development, with the
former showing much superior growth than the latter.
The positive effects observed in the current study align with the findings of
Zeynali et al., (2020) and Seyed alhosseini (2023), who found
that diets supplemented with S. ilicifolium and other seaweed species
resulted in improved development performance and feed application. Based on the
findings, Sargassum spp. Enhances growth in fish as it contains polysaccharides,
carbohydrates and bioactive compounds that improve nutritional absorption in
fish. Akbary (2019) explored that the highest body weight, condition factor and
PER were observed in Rastrelliger kanagurta at a 15% inclusion level of S.
ilicifolium.
In contrast, Shapawi and Zamry (2016) stated that Sargassum polycystum
in Asian seabass did not improve growth. Similarly, Palmaria palmata has
no significant effect on the development of Atlantic salmon Wan et al.
(2016). The inclusion of A. platensis in diets of Oncorhynchus mykiss
did not affect growth performance Teimouri et al. (2013). These
studies showed that the growth-increasing effects of seaweeds may be wary due
to different species nutritional requirements, environment, seaweed types and
dosage. Plant-based protein source for fish feed has functional benefits due to
its nutritional quality, sustainability, and cost-effectiveness. Asad et al.,
(2025).
Hematological indices, notably red blood cell count, hemoglobin, and
hematocrit values, significantly improved in the current study when Sargassum
spp. was added to fish diets at different levels (0, 3%, 5%, and 7%). These
results are consistent with earlier research by Sabzi et al. (2023),
which found that feeding fish species like L. calcarifer, Cirrhinus
mrigala, and Oreochromis niloticus diets supplemented with seaweed
significantly increased their RBC, Hb, and Hct levels.
With increasing dietary inclusion of Sargassum spp., a clear positive trend in
RBC values was shown. This was especially true for the 7% inclusion group,
which showed the highest significant rise in RBCs, Hb, and Hct when compared to
the control. The developments imply that Sargassum spp. might have bioactive
elements like polysaccharides and antioxidants, which, by enhancing oxygen
circulation and the immune system, could help fish's blood parameters and
general health.
Furthermore, the notable increase in neutrophils and WBC in diets supplemented
with Sargassum spp. in fish-eating diets is a crucial indicator of
immunological activation, which is consistent with the results of Yeganeh and
Adel (2019), who also observed elevated WBC levels. Given that the body uses
these cells to fight infections, the noted rise in neutrophils in our study
points to a better immunological response. The 7% Sargassum spp. diet shown a
notable increase in neutrophils, thus substantiating the hypothesis that
Sargassum spp. can enhance fish immune systems.
A number of investigations have found the opposite, in contrast to these
encouraging findings. A study conducted by Yang et al. (2014)
found that fucoidan, a chemical derived from brown seaweed, had no significant
effect on the red and white blood cell counts or hematocrit levels in yellow
catfish.
In conclusion, the present study confirms that dietary inclusion of Sargassum
spp. enhances hematological indices in fish, particularly RBC, Hb, Hct, and WBC
levels. These improvements in blood demonstrate that Sargassum spp. contributes
to better overall physiological status and immune function in fish. However,
some studies also showed neutral and negative effects of seaweeds.
CONCLUSION
The
findings of this study demonstrated that the growth performance and immunological
response of Labeo rohita significantly improved in the treatment groups
with the dietary inclusion of Sargassum ilicifolium than in the control
group. By comparing the treatments, the best results were observed in the
treatment with 7% sargassum inclusion (T3) followed by 5% inclusion (T2) and
the least but better than control results were observed in the treatment with
3% sargassum inclusion (T1).
Statements
and Declarations
Conflict of interest: Not applicable
Consent to Participate: All authors are agreed to
submit the article in this journal
Consent to Publish: All authors are agreed to
publish in this journal
Funding: No funding
Data availability statement: On request
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