Introduction
Genetic diversity provides the basis for aquatic species ability to withstand in ever-changing
ecological conditions by nurturing their ability to protect against extinction (Ashley et al.,2003;
Banerjee et al., 2008). To create evolutionary relationships between different species, genetic
variation is used. The genetic explanation of individual similarity and reproduction strategies is
also beneficial (Das et al., 2005). Genomic diversity is linked with a slight extent of adaptability
and can be essential for biological habitats. It makes feasible for fish to give birth to healthy
long-living offspring (Vellend and Geber., 2005). Allelic diversity must be retained in order to
manage evolutionary ability of natural populations to adapt instability of environmental
conditions and to conserve genetic wholeness (Perrier et al.,2011). Regular surveillance of the
genetic composition of the riverine fisheries stock is necessary to prevent genetic deterioration
caused by environmental changes, natural mortality, and overexploitation (Islam et al., 2005).
Scrutinizing the genetic makeup of wild fish populations furnishes important information for
creating conservation and management strategies for fish species that are list as endangered.
Insight the genetic composition of fish species that are cultivable is also fundamental for
increasing stocks, identifying possible brood stock, selecting breeding program, and managing
for a sustainable harvest and biodiversity conservation (Haniffa et al., 2007). Genetic profiling of
native populations developing due to human-induced and various ecological factors like
pesticides, chemical pollutants, agriculture run-off, and fertilizers, which are imposing additional
stress on the fish population. The primary elements that have interrupted fish species survival in
natural ecosystem over the past few decades have been human interventions, including
overfishing, the introduction of non-native species, pollution, water flow changes, and
environmental hazards like floods and climate shifts (Vandewoestijne et al., 2008). Fish species
that are commercially manipulated have substantially less genetic diversity due to inbreeding,
lack of genetic profiling, and poor management of the brood stock.
Out of the five rivers that cross the historical crossroads region of Punjab in northern India and
Pakistan, the Sutlej River is the longest. Another name for the Sutlej River is Satadru. It
functions as the Indus River's easternmost affluent. The Sutlej river in Punjab is getting
contaminated as a result of industrial effluents, household sewage, and agricultural runoff that is
usually dumped into the river or onto nearby land (Jindal and Sharma., 2011; Kaur et al., 2000;
Khurana et al., 2014; Verma et al., 2013).
The Channidae family of freshwater fish includes snakeheads. Pakistan has a representative and
diverse freshwater ecosystem with a wide variety of warm-water fish species. As a major
riverine freshwater food fish, Channa marulius, also known as snakehead locally, is widely
distributed throughout Asia (Berra et al., 2007). This fish is becoming more widespread and can
now be found in Pakistan's Azad Jammu Kashmir, Punjab, Sindh, Khyber Pakhtunkhwa, and
Balochistan provinces. Channa marulius has become increasingly important because of its high
market value, and abundant natural distribution.
C. marulius is a freshwater fish that lives in lakes, marshes, reservoirs, and expansive areas of
marshy water. It has been seen as high as 475 meters above sea level. With a length of 120–122
cm, C. marulius is the species of murrel that grows the fastest (Bardach et al., 1972; Talwar and
Jhingran, 1992). One of the largest species of snakeheads, Channa marulius (Bardach et al.,
1972), prefers deep, clear lakes and rivers with a rocky or solid sand bottom. C. marulius is often
found in association with submerged vegetation and inundated woodlands, where it constructs
vegetation nests for the purpose of spawning and raising offspring (Talwar and Jhingran., 1992).
[Link] is carnivorous and feed on tadpoles, frogs, fish, snakes, earthworms, and insects,
water birds and rodents. Males in this species are territorial (Chaudhary., 2010; Froese and
Pauly., 2017a). It has been noted that the species reproduces in almost every type of aquatic
environment, including ponds, rivers, reservoirs, rice fields, and tiny cement tanks. From such
water bodies, their progeny can be obtained in significant numbers as yolk-sac larvae,
fingerlings, fry, and eggs (floating eggs). This species builds floating nest of leaves and weeds;
having orange yellow eggs. Egg hatch time is 36-48 hours and fry stay in the nest for ten days
after hatching. Parents (C. marulius) guards the fry for a month (Chaudhary., 2010; Froese and
Pauly.,2017a).
Considered a local migrant, Channa marulius travels short distances in search of food or to
locate appropriate breeding sites in fresh bodies of water to escape the stressful conditions of its
current habitat. The fish has been assessed as being at lower risk—near threatened, or (LRnt)—
due to its declining natural abundance (CAMP et al.,1998). In recent times, there has been a
discernible decline in the natural populations of Channa marulius. This decline has been ascribed
to various factors such as habitat degradation, overexploitation, non-native species invasion,
illnesses, environmental contamination, and toxic substances.
Molecular markers have emerged as indispensable instruments in population genetics,
evolutionary biology, and conservation biology (Liu and Cordes, 2004). These markers are used
to evaluate gender-specific gene flow, historical bottlenecks, and effective population size
(Cheng et al., 2010; Kucuktas et al., 2009). However, careful marker selection helps identify the
genetic variation that matters most for any population genetic study (Sunnucks, 2000).
Microsatellites, which are also known as "simple sequence repeats," are highly polymorphic,
widely distributed DNA segments with two to nine base pairs. Most microsatellite loci are short
and simple to amplify with PCR. SSR markers are used to show relationships between people
and to generate a genetic fingerprint. Studies on fish population genetics and conservation make
extensive use of SSR markers (Dudu et al., 2015). Microsatellite DNA is one of the more
reliable molecular markers available because of its unique characteristics, which include a high
rate of mutation, a broad distribution in both coding and noncoding regions of DNA, a quick
detection protocol. (Chistiakov et al., 2006), common and flexible nature high degree of
polymorphism and their small size. (Estoup and Angers, 1998).
The most popular application of microsatellites has been in the investigation of genetic
variability in carp. Because these markers are selectively neutral, microsatellite-based data can
be reliably replicated in a variety of populations, which has led to microsatellites notable success
in population analysis. Microsatellites are especially useful in assessing genetic diversity.
Microsatellite markers are considered the best instruments for population genetics research,
genetic mapping, parentage verification, forensic profiling, and conservation biology (Jarne and
Lagoda, 1996; Peakall, Gilmore, Keys, Morgante and Rafalski, 1998). It has been shown that
sibling species and subspecies as well as geographically isolated populations can be
distinguished using microsatellite DNA markers. Microsatellites are powerful genetic markers
for genetic mapping (Sanetra et al., 2009), stock identification (Liu and Cordes, 2004), and
population differentiation because of the characteristics they possess.
LITERATURE REVIEW
Jabeen et al. (2022) examined the genetic diversity in Pakistani freshwater Channa marulius by
using microsatellite markers. In this study 150 individuals from five populations were used. With
noticed values ranging from 0.700 to 0.833, the results indicated a mediate level of
heterozygosity. The disturbance of Hardy-Weinberg equilibrium indicated the potential for
recent stock mixing or inbreeding. AMOVA indicated that within-individual genetic variation
considered for the majority (77.21%). Within the Channa marulius population, the UPGMA
dendrogram recognized two main clusters. The results of this study indicate a decline in genetic
diversity, highlighting the requirement of executing efficient management and conservation
strategies to assure the sustainability of Channa species fisheries in Pakistan.
Hussain et al. (2019) evaluated genetic polymorphism of declining wild population of Channa
marulius in Jhelum river by using microsatellite marker. A mediate degree of heterozygosity was
found using microsatellite markers, with anticipated and noticed heterozygosities of 0.54 and
0.43, respectively. Variation across populations was indicated by pairwise genetic differentiation.
AMOVA revealed significant genetic differences between populations (62.05%) instead of
within populations (33.03%). In all populations, disturbance from the Hardy-Weinberg
equilibrium proposed inbreeding. Within C. marulius populations, two major clusters were found
using the UPGMA tree and principle component analysis. For the aim of managing and
conserving fisheries along the River Jhelum, this genetic structure analysis offers crucial
knowledge.
Robert et al. (2019) studied the intra-specific diversity of Asian snakehead fish, Channa striata
found in Sabah North Borneo. In this study, six microsatellite loci were used. Evaluating the
sampled populations genetic diversity to other spots, it was mediate. The population of Sabah
was highly structured overall, reflecting isolations across ecological and geological time scales.
Along the east and west coasts, two different genodemes were found, potentially divided by
geological features. Even though there was kinship between genetic populations, there was
separate populations indicated by low gene flow and strong genotypic differentiation, which is
likely because freshwater habitats naturally fragment. The research underscores how molecular
markers can be used to define Sabah's geospatial conservation units. Two different ecoregions
that may contain allopatrically evolved biota are depicted by lowland freshwater ecosystems. In
these ecoregions, where freshwater habitat surveys have been inert, genetic research can help to
identify biodiversity hotspots, which is crucial for conservation efforts.
Pathak et al. (2018) analyzed an enriched genomic library of the great snakehead fish, Channa
marulius. The study identified 27 polymorphic loci and extracted species-specific microsatellite
markers. To ensure genetic neutrality, the repeat-containing sequences were explained. 67 people
from three Indian rivers (the Teesta, Mahanadi, and Godavari) had their genetic indices analyzed
using 19 reliable loci that were verified. Heterozygosities extent from 0.077 to 0.818, and
polymorphism information content (PIC) values were between 0.366 and 0.831. The samples
were grouped into three different populations using a Bayesian model-based clustering approach,
evaluating limited genetic admixture among them. It was founded that the validated
microsatellite markers might be useful for identifying C. marulius genetic diversity. Moreover,
C. punctata, C. gachua, and C. striata _ three related species _ were found to exhibit successful
cross-transferability. These findings provide information that can be used to manage and
conserve C. marulia.
Yan et al. (2018) examined the genetic diversity, gene flow, and population structure by utilizing
microsatellite marker among nine populations of northern snakehead fish in Central China. Small
effective population sizes carried on by overfishing and habitat loss, as well as the loss of native
genetic diversity as a result of fish movements, are the main hazards to the diversity of
freshwater fish. There is a conspicuous genetic partition based on regional and river system
separation even though there is a high level of genetic diversity. As per to the study, conservation
projects are necessary, especially for populations with small effective sizes. The improvement of
local stocks is a result of human activities such as aquaculture and the transfer of wild fish,
which features the need for creative management techniques to preserve the genetic diversity of
native freshwater fish, especially the northern snakehead.
Wegleitner et al. (2016) determined the origin of northern snakehead (Channa argus)
introductions in the eastern United States of America. The study analyzed genetic traits in
established C. argus populations, a sample from a Manhattan Chinatown market, and an Upper
Hudson River population with uncertain status. The study used four microsatellite markers in
light of the rapidly dispersing invasive species worldwide. The specimens sampled exhibited two
different genetic groups, indicating multiple introductions into U.S. waters, according to the
findings. The population of C. argus in the Upper Hudson is most analogous to the population in
the Lower Hudson near Queens, New York, in accordance to the genetic structure analysis. The
study came to the resolution that the genetic populations of the Potomac River and the
Chesapeake Bay basins are one, indicating that introductions to the Chesapeake Bay most likely
came from the Potomac population. The study also advocated that these founding populations
experienced secondary spread from humans, emphasizing the consequence of comprehending
genetic traits for efficient control of invasive species and terminate them from disseminate.
Tan et al. (2016) examined genetic variability by using nine microsatellite loci in wild striped
snakeheads (Channa striata) from Malaysia. Large effective population sizes are suggested by
the majority of populations' moderate-to-high genetic diversity. Each region's mixed, highly
diversified populations were found to be viable candidates for conservation and selective
breeding. Based on drainage patterns, three homogeneous groups of wild populations were
distinguished and kept apart by functional barriers. Long-term geographic isolation, combined
with ancient population connectivity within the same group, facilitated allele adaptation to local
environmental changes. Geographic isolation and genetic distance were found to be significantly
correlated. Anthropogenic disturbances revealed genetic similarity between geographically
separated groups, highlighting the necessity of taking into account both natural and man-made
elements in population control and conservation plans.
Zhou et al. (2015) studied nine snakehead mating groups with healthy offspring. Using five
polymorphic microsatellite loci that were chosen from isolated populations, 43 alleles with
different counts per locus were found. There was variability in the average observed and
effective allele counts per population. The amount of polymorphism information varied, as did
the heterozygosity measures (HE and HO). Notably, the Hardy-Weinberg equilibrium was
broken by 38 of the 45 loci. These discoveries are essential for evaluating and protecting new
species, providing basic information for forecasting heterosis and directing breeding initiatives.
Zhu et al. (2015) concentrated on the genetic diversity of four cultivated populations of the
Chinese food fish known as snakehead fish, or Channa argus. The study discovered high genetic
diversity with 154 alleles using ten microsatellite loci. The extent of the average noted and
awaited heterozygosities was 0.69-0.83 and 0.70-0.84, respectively. The polymorphism
information content was found to be substantially varied, ranging from 0.66 to 0.82. There were
cues of a recent bottleneck in population JX. The largest distance between the HN and SD
populations was demonstrated by pairwise genetic differentiation, which also recommended a
moderate level of variation. Genetic clustering proposed that the HN and ZJ populations most
likely shared a common ancestor. The study provides vital information about the sustainable and
conservation of Channa argus genetic resources.
Yan et al. (2014) isolated nineteen novel microsatellite loci and characterized in 42 individuals
from one natural population of Channa argus collected from Poyang Lake in Jiangxi province
The number of alleles per locus extent from 5 to 18 with an average of 11.16. The noticed and
anticipated heterozygosities extent from 0.214 to 1.000 and from 0.630 to 0.923, respectively.
The average variability detail value was 0.814. Among these polymorphic microsatellites, eleven
loci complied to Hardy–Weinberg equilibrium. These microsatellite loci are presently being used
to evaluate the genetic diversity and population structure of C. argus and will participate to the
effective conservation and rational utilization of genetic resources of this species.
Zhou et al. (2012) investigated genetic characterization of northern snakeheads population in
China. In this study five variable microsatellite were used. The data based on 260 fish from eight
populations and 95 alleles identified, including 24 private alleles. The observed alleles extent
from 5.60 to 11.40, and the effective allele numbers manifold from 3.66 to 6.67. There was
alteration in the heterozygosity values (HE and HO) between the populations (0.703 to 0.846 and
0.626 to 0.800). Most of microsatellite loci deviated from the Hardy-Weinberg balance. Five
unique clusters among populations were found by UPGMA analysis, suggesting genetic
differentiation. The study emphasizes how important it is to manage and safeguard northern
snakehead resources in order to maintain genetic diversity and stop the loss of alleles due to
overfishing in natural waters.
T.L King et al. (2011) examined northern snakehead (Channa argus) fish as a model from
Meadow Lake, New York City. The study is conducted to isolate and characterize 19 tetra-
nucleotide microsatellite DNA markers. Both allelic diversity (average 6.8 alleles/locus) and
heterozygosity (average 74.2%) were found in moderate amounts in these markers. There were
some linkage disequilibrium and divergence from Hardy-Weinberg equilibrium, which indicated
that the Meadow Lake population had not reached mutation-drift equilibrium, according to
demographic analyses. The findings suggested a configuration of multiple, recent inclusion by
showing the existence of multiple different groups of linked individuals. The C. argus
microsatellite markers that were developed showed enough genetic diversity for a range of uses,
such as defining management units for eradication efforts, defining kinship, elucidating fine-
scale population structure, estimating dispersal rates, estimating population sizes, and offering
distinct demographic perspectives on the efficacy of control or eradication measures.
Gul et al. (2010) explored Channa argus enriched genomic libraries, eleven microsatellite
markers were found. The loci of 40 specimens from a single natural population were examined.
Six to seventeen alleles were present in each locus, and the average polymorphic information
content was 0.830. The heterozygosities that were observed and predicted were 0.8000 to 1.0000
and 0.7269 to 0.9551, respectively. Three loci indicated significant linkage disequilibrium (P <
0.005), and only one locus substantially differed from the Hardy-Weinberg equilibrium. It is
expected that these recently identified markers will be essential in upcoming studies for
population genetic evaluations and the creation of a genetic linkage map for the important
snakehead fish species Channa argus.
