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Description of Three New Species of Aseptate Gregarine, Monocystis Von Stein, 1848 of Oligochaetes Collected from Dhaka, Bangladesh

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Description of Three New Species of Aseptate Gregarine, Monocystis Von Stein, 1848 of Oligochaetes Collected from Dhaka, Bangladesh Rec. zool. Surv. India: Vol. 120(3)/211–219, 2020 ISSN (Online) : 2581-8686 DOI: 10.26515/rzsi/v120/i3/2020/146821 ISSN (Print) : 0375-1511 Description of three new species of Aseptate Gregarine, Monocystis von Stein, 1848 of Oligochaetes collected from Dhaka, Bangladesh Sutapa Sarkar, Beauty Kundu and Probir K. Bandyopadhyay* Parasitology Laboratory, Department of Zoology, University of Kalyani, Kalyani – 741235, West Bengal, India; Email: [email protected] Abstract The paper deals with the description of three new species of Monocystis von Stein, 1848 namely Monocystis bangladeshensis sp. nov., Monocystis hamidae sp. nov. and Monocystis ribbonae sp. nov. obtained from the seminal vesicles of earthworm Metaphire peguana from Dhaka, Bangladesh. Trophozoites of Monocystis bangladeshensis sp. nov. elongated, measure 214.49 x 30.94 µm. Nucleus rounded, measuring 14.28 x 11.4 µm. Gametocysts ovoid, ranging 98.26 x 74.94 µm. Oocysts fusiform, measuring 15.48 x 8.85 µm. Trophozoites of Monocystis hamidae sp. nov. irregular in shape, measuring 202.41 µm in length, width of the anterior part and posterior part measure 46.41 x 19.28 µm. Nucleus rounded, measuring 6.0 µm. Gametocysts measure 150.60 µm. Oocysts rhomboid, ranging 8.00 x 5.45 µm. Trophozoites of Monocystis ribbonae sp. nov. irregular ribbon-like measuring 88.24 x 16.50 µm. Nucleus spherical, rangining 11.50 x 10.00 µm. Gametocysts ovoid, measure 59.25 x 70.37 µm. Oocysts fusiform, measuring 7.5 x 4.00 µm. Keywords: Monocystis bangladeshensis, Monocystis hamidae, Monocystis ribbonae, New Species, Taxonomy Introduction seminal vesicles of the Metaphire peguana. In this paper the description, taxonomy and systematics of three new Gregarines are a group of unicellular endoparasites species are presented. belonging to the phylum Apicomplexa under the subkingdom Protozoa. Adl et al. (2019) proposed Materials and Methods that gregarines are recognised as three major groups, archigregarines, eugregarines and neogregarines. Host earthworms were collected by one of the authors Eugregarines (Eugregarinorida Leger, 1900) divided into (SS), serving the Satkhira College, Satkhira, Bangladesh. two groups: septate gregarines and aseptate gregarines. The collected earthworms were conserving in a plastic Septate gregarines favoured in arthropods and mollusc bucket filled with soil and brought into the Parasitology and aseptate gregarines tend to harbour in oligochaetes. Laboratory, Department of Zoology, University of Kalyani. The aseptate form characterised by undivided body i.e. it The seminal vesicles were carefully removed from the doesn’t divided by septum. Whereas in the septate form dissected earthworms. Seminal vesicles were immersed body divided by septum. in 0.62 % NaCl solution. A thin film of seminal fluid has Only three species of Monocystis namely M. ayeshae been made on a slide covered with a coverslip. The slides Sarkar & Bandyopadhyay, 2011; M. sahadatae Sarkar & were examined under a phase-contrast (Olympus CX41) Bandyopadhyay, 2013 and M. apareshae Sarkar & microscope. Then the content of the seminal vesicles was Bandyopadhyay, 2014 have so far been established from semi-dried and fixed in Schaudin’s fluid for 20 minutes. Metaphire posthuma and Metaphire anomala respectively The smears were stored in 70% alcohol for removal of collected from Bangladesh. mercuric chloride. The slides were then passed through Ongoing biodiversity survey of aseptate gregarines in 100%→ 90%→ 70%→ 50% alcohol (5 minutes each) and earthworms of the Dhaka, Bangladesh revealed three new placed in distilled water. These were kept in 3% iron species of the genus Monocystis von Stein, 1848 from the alum solution (throughout the night) and stained with * Author for correspondence Article Received on: 19.08.2019 Accepted on: 19.05.2020 Description of three new species of Aseptate Gregarine... Heidenhain’s hematoxylin solution for 20 minutes. 1% Zoobank: http://Zoobank.org/urn: lsid:zoobank.org:act: iron alum solution was used for differentiation. The slides 50E4818E-3DA2-463A-8ABB-AEC5FF176433 were then washed in distilled water and dehydrated in Description: Trophozoites elongated with rounded lobe like an ascending series of alcohol. They were then cleared in an anterior end, bent into downwards forming a question xylene and mounted in Canada balsam as described by mark like structure. The posterior portion has a short tail- Bandyopadhyay et al. 2005. The Camera Lucida drawings like structure. The nucleus is rounded and present in the of the parasites have been done. The photomicrographs middle of the trophozoite. Mucorn situated at the anterior have been taken using CX41 Olympus phase-contrast end. Gametocysts are ovoid in shape (Clopton F7) and microscope. The shape of the parasites has been described oocysts are fusiform in shape (Clopton B4). following Clopton (2004). Gamonts measure 211.45 - 218.79 (214.49 ± 3.77) x 28.73 - Results and Discussion 33.15 (30.94 ± 1.88) µm. Nucleus measures 12.5 - 16.3 (14.28 ± 6.4) µm x 10.4 - 12.5 (11.4 ± 3.6) µm. Diameter Systematic Account of gametocysts measuring 94.88 - 105.66 (98.26 ± 3.69) Phylum APICOMPLEXA Levine, 1988 µm x 70.52 - 78.40 (74.94 ± 3.69) µm; Oocysts measure Class SPOROZOA Leuckart, 1879 14.27 - 17.70 (15.48 ± 1.84) µm in length and 6.7 - 11.00 (8.85 ± 1.85) µm in width. Order EUGREGARINORIDA Léger, 1900 Type material: Monocystis bangladeshensis sp.nov. Family MONOCYSTIDAE Bütschli, 1882 Type host: Metaphire peguana Rosa, 1890 Subfamily MONOCYSTINAE Bhatia, 1930 Site of infection: Seminal vesicles Genus Monocystis von Stein, 1848 Type locality: Dhaka, Bangladesh Monocystis bangladeshensis sp. nov. Prevalence: Twelve out of twenty-five host earthworms Figure 1(A-C), 2 (A-C); Table 1 and 4 (48%) were infected with the parasite. Figure 1. Monocystis bangladeshensis sp. nov. (photomi- Figure 2. Monocystis bangladeshensis sp. nov. (camera crographs of different stages of the life cycle) lucida drawings of different stages of the life A. Trophozoite; B. Gametocyst; C. Oocyst. cycle) A. Trophozoite; B. Gametocyst; C. Oo- Scale-bars. A-B, 100 µm; C, 10 µm. cyst. Scale-bars. A-B, 100 µm; C, 10 µm. 212 Vol 120(3) | 2020 | www.recordsofzsi.com Zoological Survey of India Sutapa Sarkar, Beauty Kundu and Probir K. Bandyopadhyay Material deposited: Holotype: PR/MC/27/11 is deposited W 436, Nebraska Hall, Lincoln, Nebraska 68588-0514 at the Parasitology Laboratory, Department of Zoology, USA, (Accession no. P-2014-002; Reg. no. HWML University of Kalyani, Kalyani 741235, West Bengal, India. 49902). Paratype: The paratype has been deposited in the Harold W Etymology: The specific epithet “bangladeshensis” has Manter, Laboratory of Parasitology, Systematic Research been given after the name of the country, Bangladesh Collections, University of Nebraska State Museum, from where the host specimens have been collected. Table 1. Summary of morphometric data (±µm) of the observed Monocystis bangladeshensis sp. nov. stages in seminal vesicles of Metaphire peguana (n=20) Parameters Range Mean Median SD SE CV % Length of the trophozoite 211.45-218.79 214.49 214.27 3.77 0.84 1.75 Width of the trophozoite 28.73-33.15 30.94 30.54 1.88 0.42 6.07 Length of the nucleus 12.5-16.3 14.28 14.12 6.4 1.43 44.81 Width of the nucleus 10.4-12.5 11.4 11.29 3.6 0.80 31.57 Length of the gametocyst 94.88-105.66 98.26 98.29 3.69 0.82 3.75 Width of the gametocyst 70.52-78.40 74.94 74.69 3.69 0.82 4.92 Length of the oocyst 14.27-17.70 15.48 15.38 1.84 0.41 11.88 Width of the oocyst 6.7-11.00 8.85 8.67 1.85 0.41 20.90 Table 2. Summary of morphometric data (±µm) of the observed Monocystis hamidae sp. nov. stages in seminal vesicles of Metaphire peguana (n=20) Parameters Range Mean Median SD SE CV % Length of the trophozoite 198.9-207.2 202.41 202.36 3.26 0.72 1.61 Width of anterior part of 44.20-48.62 46.41 46.31 2.26 0.50 4.86 trophozoite Width of posterior part of 17.68-19.89 19.28 19.34 0.62 0.13 3.21 trophozoite Diameter of the nucleus 5.5-6.5 6.0 6.08 0.55 0.12 9.16 Diameter of the gametocyst 145.25-160.50 150.60 150.68 6.0 1.34 3.98 Length of the oocyst 8.50-9.25 8.00 8.12 1.25 0.27 15.62 Width of the oocyst 5.25-5.50 5.45 5.35 1.21 0.27 22.20 Table 3. Summary of morphometric data (±µm) of the observed Monocystis ribbonae sp. nov. stages in seminal vesicles of Metaphire peguana (n=20) Parameters Range Mean Median SD SE CV% Length of the trophozoite 86.50 - 90.40 88.24 88.37 10.01 2.23 11.34 Width of the trophozoite 22.00 -25.00 16.50 16.34 8.20 1.83 49.69 Length of the nucleus 14.2 - 16.42 11.50 11.39 3.0 0.67 26.08 Width of the nucleus 9.1 - 11.05 10.00 10.12 1.5 0.33 15 Length of the gametocyst 55.50 - 60.25 59.25 59.19 3.8 0.84 6.41 Width of the gametocyst 67.40 -72.45 70.37 70.30 3.75 0.83 5.32 Length of the oocyst 5.5 - 9.50 7.5 7.3 1.60 0.35 21.33 Width of the oocyst 3.5 - 4.25 4.00 3.89 0.09 0.02 2.25 Vol 120(3) | 2020 | www.recordsofzsi.com Zoological Survey of India 213 Description of three new species of Aseptate Gregarine..
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