Cnidaria: Myxosporea) in India: Current Status

Published online: February 5, 2021 ISSN : 0974-9411 (Print), 2231-5209 (Online) journals.ansfoundation.org Review Article Molecular tools for identification and classification of Myxozoan parasites (Cnidaria: Myxosporea) in India: Current status Abhishek Gupta Article Info Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University, Meerut-250004 (U. P.), India https://doi.org/10.31018/ Anshu Chaudhary jans.v13i1.2451 Received: December 8, 2020 Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University, Revised: January 28, 2021 Meerut-250004 (U. P.), India Accepted: February 3, 2021 Anju Tyagi Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University, Meerut-250004 (U. P.), India Bindu Sharma* Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University, Meerut-250004 (U. P.), India Hridaya S. Singh Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University, Meerut-250004 (U. P.), India *Corresponding author. Email: [email protected] How to Cite Gupta, A. et al. (2021). Molecular tools for identification and classification of Myxozoan parasites (Cnidaria: Myxosporea) in India: Current status.. Journal of Applied and Natural Science, 13(1): 51 - 58. https://doi.org/10.31018/jans.v13i1.2451 Abstract A substantial literature on myxosporea has been published to date using morphological characters and specificity of host tissue. Currently, there are some 2400 nominal species of myxosporea classified to 17 families and 64 genera. Approximately 300 species have been described from India and most of them have been described solely on the basis of morphological characteristics. Molecular markers like small subunit ribosomal (SSU) and large subunit ribosomal (LSU) DNA have been used worldwide for the identification and validation of fish myxosporeans. Maximum likelihood phylogenetic tree based on SSU rDNA sequences was used to study the phylogenetic relationship among myxosporeans infecting Indian fishes. Myxospore phylogenies disa- gree with traditional spore-based classification systems, probably due to extreme plasticity in myxospores morphologies that have resulted in extensive convergence. Morphological similarities exist among myxosporea that encounter several problems in categorizing them. That’s why present-day research has shifted to molecular techniques for identification and correct systematics of myxosporeans. Molecular studies of myxozoans in India are still scarce and in infancy. To address persisting taxonomic and phylogenetic discrepancies, validation of these species by molecular tools is needed, because earlier species were reported only on the basis of morphological data. Therefore, the present study has summarized existing molecular data and current status of molecular taxonomy of myxosporeans parasitizing fresh and marine water fishes of India along with the approaches of myxozoan phylogenetics and information about the molecular markers, their interpretation in the identification of myxozoans parasitizing fishes. Keywords: Myxosporea, Systematics, Fish parasites, Molecular marker, Phylogenetic analysis, Validation INTRODUCTION ter as well as marine. For the first time in freshwater fish, myxozoan cysts were reported in the musculature Myxosporea representing about 20% of described cni- of Coregonus fera from a lake in Geneva by Jurine in darian species evolved as endoparasites and are radi- 1925, while in India the work on myxozoan was initiat- ated into a diverse range of morphologically different ed by Bosanquet in 1910 (Abdel-Gaber et al., 2017). A parasitic species (Fiala et al., 2015). Myxosporean par- number of myxosporean species have also been de- asites are well known for infecting fishes, both freshwa- scribed from annelids, amphibians, reptiles and birds. This work is licensed under Attribution-Non Commercial 4.0 International (CC BY-NC 4.0). © : Author (s). Publishing rights @ ANSF. Gupta, A. et al. / J. Appl. & Nat. Sci. 13(1): 51 - 58 (2021) More than 2400 species belonging to 64 genera have USE OF MOLECULAR MARKERS IN MYXOZOAN been reported worldwide (Lom and Dykova, 2006). SYSTEMATICS Studies on myxosporean parasites of fishes in India In cnidarian evolution, myxosporea derived from a com- have shown that among different genera, only Myxobo- mon ancestor and exhibited an ancient cnidarian line- lus, Henneguya and Thelohanellus have been frequent- age (Kent et al., 2001). Conservative phylogenetic ly described (Kalavati and Nandi, 2007; Kaur and markers, like small and large subunit ribosomal DNA Singh, 2012; Singh and Kaur, 2012; Kaur, 2014). Ap- gene, are required to reconstruct myxosporean diversi- proximately 300 species of myxobolids have been re- fication. Nowadays, SSU rDNA sequences are most ported till date from India (Kaur and Singh, 2012). Myx- important in the systematics of myxosporea and have obolids fauna has been investigated by a limited num- been used frequently to understand their phylogeny ber of researchers in India (Gupta et al., 2018; (Fiala, 2006; Evans et al., 2010; Zhao et al., 2013; Chaudhary et al., 2019). Many of the reported spe- Naldoni et al., 2018), due to heterogeneous nature of cies have been identified and described solely on the its conserved as well as variable regions that facilitate basis of spore morphology. The end of the 20th cen- accurate differentiation of myxosporean parasites at tury laid the foundation of the molecular era, and the different taxon levels. The conserved regions of the controversies between the traditional spore-based molecular marker are helpful in DNA sequences multi- taxonomy of myxosporeans and the rDNA-based ple alignments and also facilitate the development of phylogenies started. primers for amplification purpose while variable regions It is true that myxospore morphology remains important, give information about the diversification process. Stud- as it is the fundamental diagnostic feature for identifica- ies proved that this gene evolved very rapidly because tion, but great discrepancies were found between the SSU rDNA molecular marker of myxosporean fauna phylogenetic relationships of myxospores inferred from shows high substitution rates in their variable regions spore-based myxobolids taxonomy (Fiala and Bartoso- (Saulnier et al., 1999; Hartikainen et al., 2014). va, 2010). Morphological tools have failed to identify Large subunit ribosomal DNA (LSU rDNA) sequences congeneric species, like those found in same infection have been used for myxozoan phylogenetics due to site in different fishes with only minor differences in their conserved and variable regions. LSU rDNA molec- their morphology and structures of spores (Ye et al., ular marker has also been used in congruence with 2017). These problems have been resolved with the SSU rDNA based phylogenetic analysis and sometimes help of molecular studies (Smothers et al., 1994). used in combination for particular inferences (Whipps Smothers et al. (1994) used first time ribosomal DNA and Kent, 2006). According to several workers (Fiala (rDNA) sequence to explore the phylogenetic position 2006; Bartošová et al., 2009; Fiala and Bartosova, of the Myxozoa within Metazoa. After that, the use of 2010), the LSU rDNA sequences are more reliable than molecular data started to explain the phylogenetic SSU rDNA for myxobolids phylogeny with which we relationships within myxozoans. Different molecular also agree. However, myxosporean SSU rDNA markers, particularly small subunit ribosomal DNA sequences are more in numbers than LSU rDNA in (SSU rDNA), have been used most frequently to identi- National Centre of Biotechnology Information (NCBI) fy and study phylogeny of myxosporean parasites of database, being the first choice of marker for those fishes worldwide. Gradual enrichment of rDNA data on working in the analysis of myxozoan phylogenetics. At myxosporean species brought about the elimination intraspecies level, the internal transcribed spacer of some genera and families. At present, a consoli- region 1 (ITS 1) is found to be more reliable than SSU dated methodology consisting of spore morphology, rDNA to find the similarity or dissimilarity and related- site of sporulation, tissue as well as host explicitness ness between congeneric myxosporean species and molecular characteristics has been generally (Andree et al., 1999). Phylogenetic analysis of myxo- suggested and acknowledged for taxonomic sporean species on the basis of elongation factor (EF2) evaluation of myxosporean species and for segrega- genes was also found reliable with those based on ribo- tion of species with morphological resemblance somal DNA (Fiala and Bartosova, 2010). Molecular (Holzer et al., 2004). markers such as HSP70, mitochondrial gene 12S, COX Molecular characterization has been broadly accepted -1 and some protein-coding genes like rad51 and as a significant criterion for taxonomy and discrimina- rpl23a have also been used for molecular identification tion of congeneric myxosporean species (Bartosova et of myxosporean parasites (Whipps et al., 2004). al., 2013). There are certain ambiguities in the systematics of these parasites that need to be resolved Molecular data of Myxozoan parasites from India for future taxonomists working in this field. Therefore, According to Kalavati and Nandi (2007), approximately the aim of the present review is to summarize existing 300 species have been described from the Indian sub- molecular data of myxosporea

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