Status and Scope of Conventional Morphometry and its Integration with Bar Coding in Jammu and Kashmir (J&K) Fisheries.

Correct identification is at the core of taxonomy. Although morphometric characterization is the most straightforward and cost-effective method, it has a number of limitations. By overcoming this limitation, bar coding of mitochondrial cytochrome c oxidase gene (COI) helps in accurate and cost-effective identification of fish species. This communication discusses limitations of conventional morphometry and how its integration with bar coding can help to solve the taxonomic ambiguity of morphologically similar species. Along with that information, different water bodies from J&K region have been analysed with special reference to fish diversity which revealed that the diversity is more in Jammu region as compared to Kashmir, due to favourable climatic conditions and larger number of lotic water bodies and also the fact that the integrated approach has been successfully adopted in Kashmir valley, contributing to accurate identification of fish fauna. There is a lot of room for research in this area as bar coding of fishes has just become popular in the Jammu region, despite its immaturity. This will aid in a better knowledge of the region's fish ecology, the preservation of the gene pool, and the rise of economically significant species.


Introduction
The ichthyofauna found in riverine ecosystems provides essential supplies.Therefore, an important strategy for future sustainable use and conservation management of both the species and aquatic ecosystems is knowing the Ichthyofaunal population structure. 1cause it serves as the basis for all other life sciences, taxonomic clarity is a vital prerequisite.As sampling and identification are the initial stages, therefore it is the duty of a researcher to precisely identify a species for the purpose of conservation and sustainable use.In recent times, biodiversity research is under priority and new fish identification techniques have been developed, however, their practical application during fish identification is still in its infancy.The most popular, straightforward, economical, and historical approach of identifying fish habitats is morphometric characterization. 2,3,4rphometric Characterization All life forms express their phenotypic characters as a result of their genetic constituents and various environmental influences.Morphometrics is basically a more or less interwoven set of statistical procedures used to analyze variability in size and shape measurements of organs and organisms.Morphometric characterization is an old traditional practice being used in fisheries science since 1980's, therefore there is huge array of literature available to enhance pertinent knowledge regarding this. 5 In order to identify a species of fish, morphological characteristics such as body shape, scale size and count, colour pattern, number and relative location of fins, type and number of fin rays, and numerous relative measures of body parts are analysed. 6he statistical analysis of various morphometric characters has helped in recognition of various fish samples. 7e morphometric characterization involves the analysis of both measureable and countable characters i.e. meristic characters. 8Even though the meristic characters provide some evidence for stock separation, morphometric characters provide the best statistical separation. 9Analysis of morphometric characters help not only in correct identification of fishes but also in analysis of migration pattern, stock discrimination, reproductive isolation and commercially important species. 10though external morphological characteristics are usually used to identify fish species, there are many distinct fish species and their many developmental phases that make it challenging to do so. 11

A. Resource Specialization and Ecological Conditions of Habitat
Fish exhibit the phenomenon of morphological adaptation depending on the resources and ecological conditions of their habitat, the availability of food, the temperature etc.As a result, it is possible for the same fish raised under different ecological conditions to develop different phenotypic characteristics. 12,13,14,15,16,17hence there is a risk of misidentification if visual assessment is employed to identify fishes. 18Convergent and divergent adaptations also affect the correct identification. 19

B. Morphologically Identical Species
In certain cases using morphometry for species identification yields errornous results because of close resemblance between the morphometric characters, 20 sometimes lack of quality in original description can also lead to errornous results. 21

C. Early Life Stage
Morphological identification of eggs and larval fishes is more difficult, as their morphometric characters are not fully developed. 22,23Morphometric characters are also subject to ontogenic transformation leading to error as they change during the process of development. 24

D. Sex of the Fish
Although it has been seen that the sex does not significantly influence the morphometric or meristic characters 25 but error in morphological identification also depends on the sex of the fish eg.female sharks are more prone to misidentification as compared to male sharks, also the error rate is inversely propotional to body length. 26

E. Mislabelled Fishes
Fishes identified by conventional methods being sold in market couldbe mislabeled, either intentionally to fetch higher prices or unintentionaly due to close resemblance between species, incorrect identification of edible fishes can lead to fluctuations in market prices, also some times the mislabeled fishes being sold in the market could be poisonous. 27,28,29Therefore correct identification of fishes is essential to prevent their mislabeling. 30lso traditional methodology helps inidentifying the live or dead fish in good form, but not applicabe for identification of processed or mixed samples. 31

Variations
Even with whole fish specimens, morphometric characterization is occasionally not a good enough option because they can display either more intraspecific variations or minor interspecific variations.For example, it can be challenging to distinguish between the various Barbus species that live in the Iberian Peninsula based solely on external morphology. 32

G. lack of Classical Taxonomists and Pertinent Literature
Taxonomists provide crucial knowledge about ecosystem thereby providing the key information in life sciences.It has been estimated that about 6000-10000 taxonomists are working worldwide with only a few of them are from developing countries that inhabit most of the Earth's biodiversity. 33,34his limited taxonomic community's distribution of competence is similarly uneven; more than 80% of taxonomists are either close to or older than 50 years of age, many among which are not having much computer knowledge. 35Therefore not able to send or retrieve literature electronically, hence there is a gap in expertise, among ecologically and phylogenetically important taxon, 36 which has lead to taxonomic impediment. 37,38 India, not only do we lack an updated checklist of fishes, but also the identification keys which have not been updated after the work of Talwar and Jhingran (1991), 39 KC Jayaram(2010) 40 and Sarma and Mankodi (2017). 41Also the original descriptions are referred to forever, irrespective of the quality of the paper.Making descriptive taxonomic literature available online is still a major task to promote quality in taxonomy, the unavailability of which impacts the taxonomic process, and often leads to erroneous results and phylogenetic assumptions. 36ademics are currently researching and utilising cutting-edge identifying techniques as a result of these difficulties.The application of DNA technologies for fish identification as a potent substitute tool has overcome the limitations of morphology-based identification approaches and the lack of local fish taxonomists. 19

Dna Barcoding
Paul Hebert (2003) created the idea of DNA barcoding as a molecular identification tool, and it is now a frequently used approach for species identification even by non-specialists.Cytochrome oxidase subunit 1(COI), a mitochondrial DNA gene utilised as a universal bio-identifying system for an animal, is typically used as a short, standardised nucleotide sequence of DNA for the identification of fish in the process of DNA barcoding 42 Near the 5'end of the mitochondrial gene cytochrome c oxidase subunit 1(COI) is a 648 base pair segment known as the animal barcode region.
The idea behind this method is that even within the same species, some components of an organism's DNA would vary individually.Finding these components at the species level was the very first task for the scientists who created this method.Geographic isolation causes some populations to stop sharing genetic material, and over time, separate gene pools evolve.These sub-populations maintain morphological similarity but diverge genetically, making them unable to mate and create offspring.These species are known as cryptic species.Because of this speciation, the morphological study of these populations can become questionable as we can't be exactly sure.Such situations can be easily dealt with the molecular characterization method of DNA barcoding. 43,44hus,it has now become a widely accepted and essential method for proper identification of species on a molecular level.

WHY COI?
This method is employed because mitochondrial DNA has unique properties, such as maternal inheritance, a high copy number per cell, a lack of recombination, a lack of introns, and a greater nucleotide substitution rate, which cause variations between species to rapidly accumulate.Due to COI's low mutation rate compared to other mitochondrial genes in animals, which facilitates its recovery using polymerase chain reaction, it was also chosen as the barcode marker. 45A comparative analysis of three mitochondrial genes i.e16S rRNA, cytochrome b, and cytochrome oxidase subunit I (COI) revealed that cyt b and COI are appropriate for clear identification of fishes whereas the 16S rRNA fails to discriminate closely related fish species. 46r Coding of Fishes Globally and in India DNA barcoding finds immense application and success in fisheries and furthers the results of conventional morphometry.It is now well established and practiced all across the world.The costs involved in performing the experiment CHIB & JASROTIA, Curr.World Environ., Vol.17(3) 765-794 (2022)   were very expensive in the past but are declining with advancements in technology. 47 is commonly believed that taxonomy and barcoding compete with one another for financing, but in reality, entities other than those supporting taxonomic work fund barcoding programmes.Therefore, bar coding would not in any way compete with traditional taxonomy, and the money spent on bar coding is also used to collect and preserve specimens, which are crucial for taxonomy.Therefore, the DNA barcoding programme has the potential to significantly increase fresh financing for museums, herbaria, and individual taxonomy labs rather than reducing support for taxonomy. 48ny workers have successfully tested the methodology of barcoding in not only identifying the species but also in the discovery of new species, monitoring of fisheries quotas, correct identification of fisheries products in market, keeping a check on trade of endangered species and identification of cryptic species. 49,50,51,52,53Molecular characterization also assists in confirming the absence or presence of a species in given area. 54 India, many researchers examined Ichthyofaunal diversity using DNA barcoding as a molecular appliance both for marine as well as freshwater fishes.One of the earliest work done on bar coding of fishes in India was by Lakra et al.(2011)  55 for validating the application of bar coding.
Meanwhile many other researchers working on barcoding have confirmed its role in correct identification of fishes 56,57,58,59,60 and also that barcoding enhances the global data base for quick identification of fishes, validates the checklist of fish fauna of the area, identifies invasive species and helps in formulation of conservation strategies. 61,62ammu Region Although the region is sufficiently far west compared to the region's regular 40 to 50 mm (1.6 to 2 inches) of rainfall per month between January and March, the southern regions around Jammu typically have a monsoonal climate.Jammu town can see monthly extremes of rainfall of up to 650 millimetres in August and July, while temperatures in the warmer seasons can exceed 104°F.By early October, conditions are cool and incredibly dry, with little rain and temperatures of about 29 °C (84 °F).By September end, rainfall decreases.

Kashmir Region
The region of Kashmir is renowned for its meadows, lakes, and springs.The earliest records of the area reveal that there was once a sizable lake in the valley, which was encircled by snow-covered mountains.It is thought that Kashmir Valley was once affected by earthquakes that it split apart the mountain wall near Baramulla, letting the water from Satisar Lake pour out and leaving behind karewas, or lacustrine mud, on the mountain edges.For hundreds of millions of years, Kashmir Valley was submerged beneath the Tethya Sea, and the valley's present-day tall sedimentary rock hills were originally submerged in water.The circular but erratic Valley of Kashmir was created in this way.There are many bodies of water in this area, which has a temperate climate. 63

Fish Fauna of Kashmir Region
The importance of the fish fauna has substantially increased since the endeavour of Haeckel in 1838, 65 when he published "Fischeaus Caschmir" and thereafter various renowned ichthyologists have come up with very ingenious work like Day (1876), 66 Silas (1960), 67 Das and Subla (1964), 68 Das and Nath(1965), 69 Yousuf (1996), 70 Kullander et al.(1999), 71 Enderlin and Yousuf (1999), 72 Balkhi (2007). 73llowing table is based on compilation of most recent data obtained from details provided by numerous workers regarding the region's fish fauna as determined by conventional morphometry.
This integrated approach was also used to characterise two significant fish species from the Kashmir valley, Triplophysa marmorata and T. kashmirensis.Due to the inadequate original descriptions and the dearth of positive reviews, it is difficult to distinguish between these two species.
A morphometric and molecular analysis was carried out with this in mind.Investigation concluded that these two taxonomic Triplophysa taxa should be accepted as valid based on morphological and mt DNA COI sequence analyses.These findings can help ichthyologists better understand the ichthyofauna of the Kashmir valley and may aid them in developing methods for protecting and managing these lesser-studied native tiny species within their area of distribution 21 Water Bodies and Fish Fauna of Jammu Region Jammu region with subtropical climate is blessed with a number of lentic and lotic water water bodies offering ample water resources for development of fisheries Analysis of the data (table 2) reveals that different researchers have identified different species in the same body of water, and also that there is not a significant temporal gap between those reports, therefore taxonomic ambiguity must have been a major factor in the inconsistent results reported, as many of the species native to the region are challenging to identify using conventional morphometry.Like Identification of species of genus Schizothorax and Tryplophysa through conventional methodology can some times lead to errornous results.
The morphology of the genus Schizothorax is strikingly similar, making it challenging to distinguish between species based on morphological characteristics.
Lately collaboration of barcoding with conventional morphometry has been adopted and tested.
After performing morphometric characterization to see if barcoding can aid in accurate species identification in fishes, researchers DNA-barcoded schizothorax species from the Neelum and Jhelum rivers in Azad Kashmir.The results showed that barcoding is accurate, dependable, and has enormous potential for species identification. 84n addition to that a similar study on five different species of Schizothorax validated the role WATER BODIES OF JAMMU 64 1.Gharana Wetland

Fish Fauna of Jammu Region
Icthyofauna of the Jammu region was intensively investigated for the first time by Das and Nath  (1965,1966)  86 eventually many workers have reported fish fauna from the region like Das and Nath (1971), 87  Malhotra et al. (1975)  88 Joshi et al. (1978), 89 Tilak (1971) 90 Dutta and Malhotra(1984), 91 Jyoti et al. (2006) 92 and Balkhi (2007). 73ollowing table is based on a collection of recent information on fish diversity provided by multiple workers for various lentic and lotic water bodies in the Jammu region, as determined by conventional morphometry.

Status of Bar Coding in Jammu Region
The present study, which is based on an examination of past findings, found that several lentic and lotic water bodies in the area has documented about 160 species.However, due to a lack of molecular characterization work, the employment of contemporary methodologies is still in its infancy.
Arif and Gandotra (2017) 110 carried out DNA barcoding of ornamental fishes in various water bodies in the Jammu region for the first time, verifying its usage for precise species identification.Analysis of the economic value of the fish fauna of the region reveals that the majority are food fishes, with ornamental fishes accounting for the secondhighest percentage, and the remainder have both food and ornamental fishes as economic status.
Since food and forage fish make up the majority of the fish in the area, molecular characterization of the major section of fish diversity remains untouched, placing the data available regarding the current status of fish fauna under uncertainty.
As shown by the data in table 5, the Jammu region is blessed with a rich diversity of fish species, many of which are generally difficult to identify morphometrically.However, bar coding is still in its infancy in this area.Major factors contributing to this research gap include outdated knowledge of current techniques, lack of funding, greater expertise in conventional methodologies, and most importantly, the fact that basic research is being neglected in favour of applied research as taxonomy has taken a backseat over time.Because most of the species lack accurate taxonomic resolution, analyses of the historical record of fish distribution, making temporal comparisons, and tracking the proper phylogeny have all been impeded.

Discussion
Due to the difference in topography, larger number of lotic water bodies, and more favourable climatic conditions, the fish diversity in the Jammu region is greater than that in the Kashmir region.
Cypriniformes is the most prevalent order in both Jammu and Kashmir due to their great level of adaptability and capacity to occupy any area.Along with the endemic species of the genus Schizothorax, many of the fishes listed above (tables 4 and 5) are exotic species, such as carps, which are not native to the area and were introduced by the state fisheries department.

Anthropogenic Stress, Declining Fish Diversity and Need for Conservational Measures F i s h , w h i c h h a v e a h e t e r o t h e r m i c b o d y
temperature, are easily impacted by changes in the physicochemical characteristics of the body of water they reside in. 111,112The aquatic ecosystem is being negatively impacted by climate change and anthropogenic factors such as pollution, overfishing, hydropower projects, etc.These factors also cause coral bleaching, the loss of coastal wetland, changes in the distribution and timing of freshwater flow, and a decline in fish diversity. 113e largest freshwater lake in Asia, Wular Lake, is home to several fish species.However, eutrophication caused by human activity, which alters the water's physicochemical properties and impairs ecological conditions, has caused the extinction of numerous schizothorax species that are adapted to clean water. 79,80Fish population of Schizothorax plagiostomus and Schizothorax esocinus in Dal lake has also been affected because of the constantly degrading water quality of the water body. 114e River Jhelum, a significant tributary of the Indus River System that drains through the entire state of Kashmir, is a celebrated river economically and a significant source of water for expanding human population and irrigation.However, the water body's shifting biological conditions have also encouraged the effective colonisation of foreign fish species with exceptional adaptability.The Viashaw River, a left tributary of the Indus River System, is also being affected by illicit mining and overfishing, which is reducing the diversity of fish. 83,114g. 4: Showing percentage of explored and unexplored water bodies of Jammu region.
Water pollution has affected the fish fauna of Jammu region as well, a comparative analysis of fish fauna of different water bodies have revealed a decline in fish diversity, 96 especially those of threatened species. 115Therefore for conservation of the river system allochthonous sources of pollution like sewage, dumping of garbage, mining and agricultural activities needs to be monitored.Different conservation measures like using a species as flagship species,creating awareness and starting different projects towards conservation needs to be adopted, 116 also small hydropower projects should be prioritised over large reservoir-based hydropower projects since they are more environmentally friendly and have fewer negative effects on flora and wildlife. 117

Research Gap and Future Prespective
Only a small number of the area's waterbodies have been thoroughly examined; the remainder of the wetlands and many lentic water bodies have mostly remained uncharted due to accessibility concerns (remote location), financial limitations, and the locals' intense religious convictions.Fish production in the area can increase significantly when the existing resources (waterbodies and fish fauna) are used wisely in order to meet the demands of UT's growing population.As the majority of the water bodies in the Jammu and Kashmir region are unexplored, concealing a substantial portion of the fish flora and its gene pool, an integrative strategy can assist close the study gap.
Therefore a collaboration on the molecular aspect of fisheries in J and k especially in Jammu region with conventional taxonomy will immensely help in better understanding of the fish ecology of the region and will also aid in properly identifying and conserving the gene pool thereby boosting the growth of the economically important fishes belonging to this region.

Fig 1 :
Fig 1: Displaying the GIS-marked Lentic and Lotic water bodies in the Kashmir region.

Fig. 3 :
Fig. 3: Percentage of explored and unexplored water bodies of Kashmir region (based on information from tables 1 and 2).Current Status of Barcoding Integrated with Conventional Morphometry in the Valley.Analysis of the data (table2) reveals that different researchers have identified different species in the same body of water, and also that there is not a significant temporal gap between those reports, therefore taxonomic ambiguity must have been a major factor in the inconsistent results reported, as many of the species native to the region are challenging to identify using conventional morphometry.Like Identification of species of genus Schizothorax and Tryplophysa through conventional methodology can some times lead to errornous results.

Fig. 2:Percentage contribution of different orders to fish diversity of Kashmir region (according to table 3's data)
Analysis of the data in table 2 demonstrates that only a small portion of the total number of water bodies existent (given in table1) have been extensively studied, leaving the majority of water bodies undiscovered.

Table 5 : Status of fish fauna reported from the Jammu region
According to an analysis of the data in table 5, only a small percentage of the total number of water bodies that are known to exist (as shown in table4)have undergone comprehensive research, leaving the bulk of water bodies unexplored.