Skip to main content Skip to main navigation menu Skip to site footer
Type: Article
Published: 2023-03-15
Page range: 93-100
Abstract views: 224
PDF downloaded: 36

New Earthworm Record from Division Muzaffarabad, Azad Kashmir, Pakistan Supported by Molecular Markers

1Microbial Biotechnology and Vermitechnology laboratory, Department of Zoology, University of Azad Jammu and Kashmir, King Abdullah Campus, Chattar Kalas, Muzaffarabad, 13100, Pakistan.
1Microbial Biotechnology and Vermitechnology laboratory, Department of Zoology, University of Azad Jammu and Kashmir, King Abdullah Campus, Chattar Kalas, Muzaffarabad, 13100, Pakistan.
2Departamento de Ecoloxia e Bioloxia Animal, Universidade de Vigo, Vigo, E-36310, Spain.
3Department of Computer Sciences and Information Technology, University of Azad Jammu and Kashmir, King Abdullah Campus, Chattar Kalas, Muzaffarabad, 13100, Pakistan.
4Department of Zoology, Government College University Lahore, Pakistan.
Departamento de Biodiversidad, Ecología y Evolución, Facultad de Ciencias Biológicas, Universidad Complutense de Madrid, Madrid, Spain.
Annelida Azad Kashmir earthworms Crassiclitellata Perelia integrative systematics molecular barcoding


Earthworm diversity and ecology in Pakistan is poorly known, especially in the region of Azad Jammu & Kashmir. An earthworm community survey assisted by genetic barcoding detected an unidentified species which could constitute a new record for Pakistan. Morphological study revealed its identity as Perelia kaznakovi. Additionally, Bayesian phylogenetic inference based on five mitochondrial and nuclear molecular markers was performed. Results provided a phylogenetic placement of the genus Perelia within Lumbricidae for the first time, indicating a close relationship with Eophila. This approach should be implemented to Perelia arnoldiana and further representatives of the genus in order to understand their biogeography, diversity and evolutionary history.



  1. Andleeb, S., Ejaz, M., Awan, U.A., Ali, S., Kiyani, A., Shafique, I. & Zafar, A. (2016) In vitro screening of mucus and solvent extracts of Eisenia foetida against human bacterial and fungal pathogens. Pakistan Journal of Pharmceutical Science, 29, 969–977.
    Asirovic, H.B. (2011) Studying the importance of the Kashkadarya oasis on the forest ecology and the increase of the soil fertility. Karshi State University, Ministry of National Education, Master academic degree dissertation, 70 pp.
    Blakemore, R.J. (2008) An updated list of valid, invalid and synonymous names of Criodriloidea and Lumbricoidea (Annelida: Oligochaeta: Criodrilidae, Sparganophilidae, Ailoscolecidae, Hormogastridae, Lumbricidae, Lutodrilidae). In: Ito M.T. & Kaneko N, (Eds.), A series of searchable texts on earthworm biodiversity, ecology and systematics from various regions of the world. Yokohama: Yokohama University, 1–80.
    Blouin, M., Hodson, M.E., Delgado, E.A., Baker, G., Brussaard, L., Butt, K.R., Dai, J., Dendooven, L., Peres, G., Tondoh, J.E., Cluzeau, D. & Brun, J.J. (2013) A review of earthworm impact on soil function and ecosystem services European Journal of Soil Sciences, 62 (4), 161–82.
    Briones, M.J.I., Morán, P. & Posada, D. (2009) Are the sexual, somatic and genetic characters enough to solve nomenclatural problems in lumbricid taxonomy? Soil Biology & Biochemistry, 41, 2257–2271.
    Brown, G.G. (1995) How do earthworms affect microfloral and faunal community diversity? Plant Soil, 170 (1), 209–31.
    Brown, W.M., George, M.J. & Wilson, A.C. (1979) Rapid evolution of animal mitochondrial DNA. Proc. Natl. Acad. Scienc, USA, 76, 1967–1971.
    Chang, C.H., Rougerie, R. & Chen, J.H. (2009) Identifying earthworms through DNA barcodes: Pitfalls and promise. Pedobiologia, 52, 171–80.
    Csuzdi, Cs., Pavlíček, T. & Mýsýrlýoðlu, M. (2007) Earthworms (Oligochaeta: Lumbricidae, Criodrilidae and Acanthodrilidae) of Hatay Province, Turkey, with description of three new lumbricids. Acta Zoologica Academiae Scientiarum Hungaricae, 53 (4), 347–361.
    Csuzdi, Cs. & Zicsi, A. (2003) Earthworms of Hungary (Annelida: Oligochaeta; Lumbricidae). Hungarian Natural History Museum, Budapest.
    Csuzdi, Cs. & Pavlíček, T. (2005) Earthworms from Israel. II. Remarks on the genus Perelia Easton, 1983 with description of a new genus and two new species. Acta Zool. Acad. Sci. Hungaricae, 51, 75–96.
    Csuzdi, Cs., Pavlíček, T. & Mýsýrlýoðlu, M. (2007) Earthworms (Oligochaeta: Lumbricidae, Criodrilidae, Acanthodrilidae) of the Hatay Province, Turkey, with description of three new lumbricids. Acta zool. Hung, 53, 347–361.
    Decaëns, T., Porco, D., Rougerie, R., Brown, G.G. & James, S.W. (2013) Potential of DNA barcoding for earthworm research in taxonomy and ecology. Applied Soil Ecology, 65, 35–42.
    Decaëns, T., Porco, D., James, S.W., Brown, G.G., Chassany, V., Dubs, F., Dupont, L., Lapied, E., Rougerie, R., Rossi, J.P. & Roy, V. (2016) DNA barcoding reveals diversity patterns of earthworm communities in remote tropical forests of French Guiana. Soil Biology and Biochemistry, 92, 171–183.
    Domínguez, J., Aira, M., Porto, P.G., Díaz-Cosín, D.J. & Pérez-Losada, M. (2018) Multigene phylogeny reveals two new isolated and relic earthworm genera (Oligochaeta: Lumbricidae). Zoological Journal of the Linnean Society, 182 (2), 258–274.
    Domínguez, J., Aira, M., Breinholt, J.W., Stojanovic, M., James, S.W. & Perez-Losada, M. (2015) Underground evolution: new roots for the old tree of lumbricid earthworms. Molecular Phylogenetics and Evolution, 83, 7–19.
    Doube, B.M., Stephens, P.M., Davoren, C.W. & Ryder, M. H. (1994) Interactions between earthworms, beneficial soil microorganisms and root pathogens. Applied Soil Ecology, 1, 3–10.
    DriloBASE: World Earthworm Database. Drilobase Project. 2014-. Available from: (Accessed 2 February 2019)
    Edwards, C. A. & Bohlen, P.J. (1996) Biology and ecology of earthworms. London: Chapman and Hall.
    Edwards, C.A. (2004) Earthworm Ecology, second ed. CRC Press, Boca Raton, Florida.
    Edwards, C.A. & Bohlen, P.J. (1996.)Biology and Ecology of Earthworms. Chapman & Hall, London.
    Farhadi, Z., Malek, M. & Elahi, E. (2013) Review of the earthworm fauna of Iran with emphasis on Kohgiluyeh & Boyer-Ahmad Province. Zootaxa, 3670 (4), 440–8.
    Fernández, R., Novo, M., Marchán, D. F. & Díaz-Cosín, D.J. (2016) Diversification patterns in cosmopolitan earthworms: Similar mode but different tempo. Molecular Phylogenetic and Evolution, 94, 701–8.
    Folmer, O., Black, M., Hoeh, W., Lutz, R. & Vrijenhoek, R. (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology, 3 (5), 294–299.
    Fushitani, K. & Riggs, A. F. (1991) The extracellular hemoglobin of the earthworm Lumbricus terrestris. Oxygenation properties of isolated chains, trimer, and a reassociated product. Journal of Biological Chemistry, 266, 10275–10278.
    Gates, G.E. (1975) Contributions to a revision of the earthworm family Lumbricidae. XII. Enterion mammale Savigny, 1826 and its position in the family, Megadrilogica, 2 (1975) 1–5.
    Ghafoor, A. & Qureshi, J. I. (1999) Five new records of earthworms from Faisalabad, Pakistan. International Journal of agricultural Biology, 1, 74–75.
    Hebert, P.D.N., Cywinska, A., Ball, S.L. & de-Waard, J.R. (2003) Biological identifications through DNA barcodes. Proc Roy Soc London, B. Biol. Science, 270, 313–21.
    Jalal, F., Rana, S.A., Qureshi, J.I., Bukhari, S., Haque, A. & Khlid, Z. M. (2014) The behavior of earthworm populations and their effect in the soil fertility with respect to climatic factors in the selected fields. International Journal of Agriculture and Crop Sciences, 7 (12), 1246-1253.
    James, S.W. & Davidson, S.K. (2012) Molecular phylogeny of earthworms (Annelida: Crassiclitellata) based on 28S, 18S and 16S gene sequences. Invertebrate Systematics, 26, 213–229.
    King, R.A., Tibble, A.L. & Symondson, W. O. (2008) Opening a can of worms: unprecedented sympatric cryptic diversity within British lumbricid earthworms. Molecular Ecology, 17, 4684–4698.
    Klarica, J, Kloss-Brandstätter, A., Traugott, M. & Juen, A. (2012) Comparing four mitochondrial genes in earthworms – implications for identification, phylogenetics, and discovery of cryptic species. Soil Biology and Biochemistry, 45, 23–30.
    Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. (2018) MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution, 35, 1547-1549.
    Lavelle, P. & Spain, A.V. (2001) Soil Ecology. Kluwer Academic Publishers, Dordrecht, The Netherlands, 2001.
    Marchán, D. F., Decaëns, T., Domínguez, J. & Novo, M. (2022). Perspectives in Earthworm Molecular Phylogeny: Recent Advances in Lumbricoidea and Standing Questions. Diversity, 14 (1), 30.
    Michaelsen, W. (1900) Oligochaeta (Vol. 10). R. Friedländer und Sohn.
    Mirmonsef, H., Malek, M. & Latif, R. (2011) The Earthworm Fauna of Tehran Province, Iran: an Ecological Characterization. Iranian Journal of Animal Biosystematics, 7 (2), 89–97.
    Paoletti, M.G., Sommaggio, D. & Fusaro, S. (2013) Proposta di Indice di Qualità Biologica del Suolo (QBS-e) basato sui Lombrichi e applicato agli Agroecosistemi Biologia Ambientale, 27 (2), 25–43.
    Pavlíček, T. & Csuzdi, C. (2006) Species richness and zoogeographic affinities of earthworms in Cyprus. European Journal of Soil Biology, 42, S111–S116.
    Pelosi, C., Bertrand, M., Capowiez, Y., Biozard, H. & Estrade, J.R. (2009) Earthworm collection from agricultural fields: Comparisons of selected expellants in oresence/ absences of hand-sorting. European Journal of Soil Biology, 45, 176–183.
    Pérez-Losada, M., Breinholt, J.W., Aira, M. & Domínguez, J. (2015) An updated multilocus phylogeny of the Lumbricidae (Annelida: Clitellata: Oligochaeta) earthworms. Journal of Phylogenetics & Evolutionary Biology, 3, 1.
    Pérez-Losada, M., Ricoy, M., Marshall, J.C. & Domínguez, J. (2009) Phylogenetic assessment of the earthworm Aporrectodea caliginosa species complex (Oligochaeta: Lumbricidae) based on mitochondrial and nuclear DNA sequences. Molecular Phylogenetics and Evolution, 52 (2), 293–302.
    Perez-Losada, M., Breinholt, J.W., Porto, P. G., Aira, M. & Dominguez, J. (2011) An earthworm riddle: systematics and phylogeography of the Spanish lumbricid Postandrilus. PLoS One, 6, 28153.
    Pop, A.A., Wink, M. & Pop, V.V. (2003) Use of 18S, 16S rDNA and cytochrome c oxidase sequences in earthworm taxonomy (Oligochaeta, Lumbricidae). Pedobiologia, 47, 428–433.
    Qiu, J.P. & Bouché, M. (1998a) Révision du genre Prosellodrilus Bouché, 1972 (Oligochaeta: Lumbricidae); description de 16 taxons nouveaux pour la science. Documents pédozoologiques et integrologiques, 4 (5), 37–64.
    Qiu, J.P. & Bouché, M. (1998b) Contribution à la taxonomie de Zophoscolexini (Oligochaeta).
    Qiu, J.P. & Bouché, M. (1998c) Révision des taxons supraspécifiques de Lumbricoidea. Documents pédozoologiques et integrologiques, 3 (6), 179–216.
    Qiu, J.P. & Bouché, M. (1998d) Révision morphologique, chorologique et taxonomique du genre Scherotheca Bouché, 1972 (Oligochaeta: Lumbricidae). Doc. pédozool. integrol, 4, 117–139.
    Rakhmatullaev, A., Gafurova, L. & Egamberdieva, D. (2010) Ecology and role of earthworms in productivity of arid soils of Uzbekistan. Dynamic Soil, Dynamic Plant, 4 (1), 72–75.
    Raw, F. (1959) Estimating earthworm populations by using formalin. Nature, 184, 1661–1662.
    Saitou, N. & Nei, M. (1987) The neighbor-joining method—a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 1987 (4), 406–425.
    Sechi, V., Brussaard, L., De-Goede, R.G.M., Rutgers, M. & Mulder, C. (2015) Choice of resolution by functional trait or taxonomy affects allometric scaling in soil food webs. Am. Nature, 185, 142–149.
    Shen, H.P., Tsai, C.F. & Tsai, S.C. (2003) Six new earthworms of the genus Amynthas (Oligochaeta: Megascolecidae) from central Taiwan. Zoological Studies, 42, 479–490.
    Sims, R.W. & Gerard, B.M. (1985) Earthworms. Keys and notes for the identification and study of the species. Bath: The Pitman Press, 1985, 171 p.
    Stephenson, J. (1930) The Oligochaeta. Oxford, Clarendon Press, pp. 1-978.
    Szederjesi, T., Pavlíček, T. & Csuzdi, C. (2013a) New data to the earthworm fauna of Israel (Oligochaeta, Lumbricidae). Opuscula zoologica Budapest, 44, 201-209.
    Szederjesi, T., Pavlíček, T. & Csuzdi, C. (2016) Description of the first endemic earthworm species from Cyprus (Oligochaeta: Lumbricidae). Zoology in the Middle East, 62, 158–163.
    Tamura, K., Nei, M. & Kumar, S. (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proceedings of the National Academy of Sciences (USA), 101, 11030-11035.
    Tsai, C.F., Tsai, S.C. & Liaw, G.J. (2000) Two new species of protandric pheretimoid earthworms belonging to the genus Metaphire (Megascolecidae: Oligochaeta) from Taiwan. Journal of Natural History, 34, 1731–1741.
    van Groenigen, J.W., Lubbers, I.M., Hannah-Vos, M.J., Brown, G.G., Gerlinde, B., De-Deyn. & van Groenigen. K.J. (2014) Earthworms increase plant production: a meta-analysis. Scientific report, 4, 6365.