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Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology logoLink to Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology
. 2011 Aug 7;35(2):169–176. doi: 10.1007/s12639-011-0061-4

Two new species of Myxobolus (Myxozoa: Myxosporea: Bivalvulida) infecting an Indian major carp and a cat fish in wetlands of Punjab, India

Harpreet Kaur 1,✉, Ranjeet Singh 1
PMCID: PMC3235390  PMID: 23024499

Abstract

A survey of parasites of freshwater fishes in Kanjali and Harike wetlands of Punjab (India) revealed the presence of two new myxosporean species belonging to the genus Myxobolus Bütschli (Winter (ed): Protozoa, 1882). Spores of the first species, M. saugati sp. nov. parasitizing scales of Labeo rohita (Cypriniformis: Cyprinidae) vern. rohu are characterized in having ‘spores measuring 8.3 × 6.6 μm in size, oval to spherical in valvular view having rounded anterior and posterior ends; polar capsules are two, equal, measuring 4.0 × 2.4 μm, pyriform with distinct neck at the anterior end; an intercapsular process is absent’. Spores of the second species, Myxobolus szekeli sp. nov. parasitizing internal wall of stomach of Wallago attu (Cypriniformis: Siluridae) vern. mulle are ‘8.7 × 4.1 μm in size, elongately ovoidal in valvular view having tapering, bluntly pointed anterior end and rounded posterior end; polar capsules are two, equal, measuring 4.4 × 1.5 μm, tubular with blunt anterior and rounded posterior ends; polar capsules converge anteriorly but are placed at a distance posteriorly and occupies nearly half of the spore body cavity; an intercapsular process is absent’.

Keywords: Cypriniformes, Harike wetland, Kanjali wetland, L. rohita, W. attu

Introduction

Kanjali and Harike are major Wetlands spread over 185 and 4,100 ha area respectively in Punjab, India among other Wetlands (i.e. Ropar, Ranjit Sagar and Nangal wetlands) in the Ramsar list Wetlands of International importance. Harike wetland harbour 26 species and Kanjali wetland 17 species of freshwater fishes. These large varieties of fishes in these wetlands are vulnerable to various parasitic infections, out of which Myxozoa is emerging as the major group. Presently, there are more than 2,180 species in 62 genera belong to the Phylum Myxozoa has been described from fishes (Lom and Dykova 2006). Recently a new genus, Thelohanelloid bengalensis from gall bladder of Arius sagor (a marine fish in Bay of Bengal) has been described by Sarkar (2009). Out of this, 744 species belong to the genus Myxobolus and is the most common genus reported worldwide (Eiras et al. 2005). According to Kalavati and Nandi (2007) 104 species of Myxobolus spp. are restricted to the Indian sub-continent. Recently, Kaur and Singh (2008, 2009, 2010a, b, 2010/2011, 2011a, b, c, d, e) have contributed 15 new species to the genus Myxobolus from freshwater fishes in Wetlands of Punjab.

During the present investigation 62 fishes belonging to L. rohita and 25 fishes belonging to W. attu were examined to ascertain the prevalence of myxozoan parasites in these wetlands. A variety of other freshwater fishes were also collected and examined which include Cyprinus carpio, Catla catla, Amblypharyngodon mola, Labeo bata, Labeo dero and Mystus seenghala. Two new species M. saugati and M. szekeli are reported from scales and internal wall of stomach respectively. The description has been prepared according to the guidelines of Lom and Arthur (1989).

Materials and methods

Fishes collected from the Kanjali and Harike wetlands were brought to the laboratory and examined for myxozoan infections. Plasmodia when found were removed and teased on slide and covered with cover slip and examined under the oil immersion for the presence of myxospores. Fresh spores were treated with 8% KOH solution for the extrusion of polar filaments. For permanent preparation, air-dried smears were stained with Ziehl–Neelsen and iron–haematoxylin. Drawings were made from stained material with the aid of camera lucida. Measurement of spores was done with the aid of a calibrated ocular micrometer. All measurements are presented in μm as range values followed by mean ± SD in parentheses.

Results and discussion

M. saugati sp. nov.

Plasmodia

Large, white to pale yellow, present all over the scales, 10–12 in number and measure 1.0–1.5 mm in diameter. 15–18 Spores are present per plasmodium.

Spore description (Figs. 1a–c, 2a, b, 5a; Table 1) (measurements based on 8–9 spores in frontal view)

Fig. 1.

Fig. 1

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a Spore stained in Ziehl–Neelsen (valvular view). b Spore stained in iron–haematoxylin. c Spore in side view. Scale bar 0.005 mm

Fig. 2.

Fig. 2

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a Spores stained in Ziehl–Neelsen. b Spore stained in iron–haematoxylin (extructed polar filaments)

Fig. 5.

Fig. 5

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a Fresh spore of M. saugati sp. nov. b Fresh spores of M. szekeli sp. nov.

Table 1.

Measurements (in μm) and ratio of M. saugati sp. nov.

Characters Range Mean values SD
LS 7.9–8.7 8.3 0.56
WS 6.4–6.8 6.6 0.28
LPC 3.7–4.3 4.0 0.42
WPC 2.0–2.8 2.4 0.56
Ratio: LS/WS 1.2
ICP Absent
NC 4–5
Parietal folds Absent
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LS length of spore, WS width of spore, LPC length of polar capsule, WPC width of polar capsule, ICP intercapsular process, NC number of coils of polar filaments, SD standard deviation

The spores are histozoic, measure 8.3 × 6.6 μm, oval to spherical in valvular view having rounded anterior and posterior ends. Two shell valves are thick, smooth, symmetrical and measure 0.6 μm in thickness. Parietal folds are absent. Polar capsules are two, equal, measure 4.0 × 2.4 μm and are pyriform with distinct neck at the anterior end. They converge anteriorly and are placed at a distance posteriorly. Polar filaments form 4–5 coils and are arranged obliquely to the polar capsule axis. They are equal when extruded and measure 26.6 μm in length. An intercapsular process is absent. Two capsulogenic nuclei are present beneath each polar capsule measuring 1.50 μm in diameter. Sporoplasm agranular, homogenous occupying whole of the extracapsular space behind the polar capsules and contain one nucleus and a large iodinophilous vacuole measuring 1.6 and 2.5 μm in diameter respectively.

Taxonomic summary of M. saugati sp. nov.

 

Type host Labeo rohita vern. rohu
Type locality Kanjali wetland, Punjab, India
Type specimen Paratypes are spores stained in Ziehl–Neelsen and iron–haematoxylin, deposited in the museum of department of Zoology, Punjabi University, Patiala. Slide no. CM/P/ZN/21.04.2009 and CM/P/IH 11.08.2009.
Site of infection Scales
Prevalence of infection 56.4% (35/62)
Etymology The species epithet saugati has been given after the name of the Dr. Saugata Basu, an eminent worker of Protozoology Laboratory, Department of Zoology, University of Kalyani, Kalyani, West Bengal, India.
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Discussion

The present species under study was compared with M. pfeifferi, M. minutus, M. catostomi, M. catlae, M. sangoricus, M. baueri, M. lotae, M. strelkovi, M. bhadrensis, M. shetii, M. rohitae, M. gobiorum, M. dermatis, M. haldari, M. cartilaginis, M. serrata, M. kwangsiensis, M. nkolyaensis, M. edellae, M. shantipuri, M. chilkensis, M. tingrelaensis, M. sushmii, M. punjabii and M. patialensis but differs from all of the above species in morphometrics (Table 2).

Table 2.

Comparative description of M. saugati sp. nov. with morphologically similar species (measurements are in micrometer)

Species Host Site of infection Locality Spore Polar capsule
M. saugati sp. nov. (present study) Labeo rohita Scales Kanjali wetland Punjab (India) 8.3 × 6.6 4.0 × 2.4
M. pfeifferi Thelohan 1895 Barbus barbus Kidney, spleen, muscles, gills – 10.0–13.0 × 9.0–12.2 5.0–5.7
M. minutus Nemeczek 1911 Leuscus leuscus, L. cephalus Gills Germany 6.0 × 4.2–5.0 3.0 × 2.0
M. catostomi Kudo 1923 Catastomus commersonii Muscles Canada 10.0–12.7 × 7.6–10.0 3.6–5.5 × 1.3–2.7
M. catlae Chakravarty 1943 L. rohita Gills West Bengal (India) 14.5–16.5 × 6.18 10.3–12.3 × 2.06–3.01
M. sangoricus Gogebashvili 1966 Varicorhynus capoeta Fin, gills, kidney Caucasus 9.1–11.0 × 7.8–9.2 3.1–3.9 × 1.8–3.0
M. baueri Chernova 1970 Tinca tinca Gills, heart, kidney, liver Russia 14.0–16.0 × 9.3–12.0 5.3–6.0 × 2.7–3.3
M. lotae Mitenev 1971 Lota lota Gills Russia 8.4–10.4 × 6.2–6.5 3.4–4.4 × 2.3–3.4
M. strelkovi Kostarev and Kulemina 1971 Phoxinus phoxinus, Leuciscus idus Gills, liver Russia 8.0–12.2 × 6.0–11.0 3.3 × 5.4 × 2.0–4.1
M. bhadrensis Seenappa and Manohar 1981 L. rohita Muscle Karnataka (India) 9.5 × 7.14 3.5 × 2.2 and 2.5 × 1.75
M. shetti Seenappa and Manohar 1981 Cirhina mrigala Gills Karnataka (India) 8.8 × 7.4 3.4 × 2.3
M. rohitae Haldar et al. 1983 L. rohita, L. bata, L. dyocheilus, L. calbasu, Puntius sarana Scales West Bengal (India) 10.6 × 9.0 6.6 × 3.3
M. gobiorum Donec 1984 in Shulman Gobio gobio Fin Ukraine 11.0–13.0 × 9.0–10.0 5.2–6.5 × 2.2–3.0
M. dermatis (Haldar et al. 1981) Gupta and Khera 1988 L. rohita Scales West Bengal Kerala (India) 10.3 × 9.4 4.4 × 2.2
M. haldari Gupta and Khera 1989 L. rohita Fin, gills Punjab (India) 9.31 × 7.95 4.31 × 2.97 and 2.95 × 1.98
M. cartilaginis (Hoffman et al. 1965) Landsberg and Lom 1991 Lepomis macrochirus Head cartilage USA 10.2 × 8.9 5.3 × 3.3
M. serrata Pagarkar and Das 1993 Cyprinus carpio Gill arch West Bengal (India) 9.07 × 8.5 4.47 × 2.6
M. kwangsiensis Hsieh et al. 1993 L. rohita Gills, kidney China 10.9 × 6.9 5.5 × 2.9
M. nkolyaensis Fomena and Bouix 1994 Barbus jae Caudal muscles Cameroon 9.0 × 8.3 4.4 × 3.0
M. edellae Sarkar 1999 Ctenopharyngodon idella Kidney West Bengal (India) 10.7 × 9.4 6.14 × 3.12
M. shantipuri Basu and Haldar 2002 Catla catla × L. rohita Gill lamellae West Bengal (India) 7.3 × 5.8 4.0 × 2.4
M. tingrelaensis Boungou et al. 2006 Sarotherodon galilaeus Among the rays of fin West Africa 11.6 × 9.37 4.80 × 2.40
M. chilkensis (Kalavati et al. 1992) Kalavati and Nandi 2007 L. rohita Gall bladder Orissa (India) 7.2–8.0 (7.77) × 5.6–6.6 (6.2) 3.2–4.8 (4.2) × 1.8–2.2 (1.98) and 1.0–1.2 (1.08)
M. sushmii Kaur and Singh 2010/2011 L. rohita Eye ball Harike wetland, Punjab (India) 9.0–10.2 (9.6) × 7.3–9.3 (8.3) 4.1–5.3 (4.70) × 2.9–3.5 (3.20) and 2.9–3.4 (3.10) × 1.0–3.0 (2.0)
M. punjabii Kaur and Singh 2010/2011 L. rohita Caudal fin Kanjali Wetland Punjab (India) 8.1–9.7 (8.9) × 6.4–7.5 (6.9) 3.4–5.4 (4.4) × 2–3.4 (2.7) and 2.8–3.6 (3.2) × 1.2–2.2 (1.7)
M. patialensis Kaur and Singh 2011e L. rohita Caudal fin Ropar wetland, Punjab (India) 11.28 × 6.67 4.8 × 3.1 and 1.70 × 1.51
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The present species is oval to spherical in shape with rounded anterior and posterior extremities without any parietal folds. They contain equal, pyriform polar capsules with anterior ends terminating into a distinct neck. In this respect, it is comparable with the spores of M. minutus, M. catostomi, M. sangoricus, M. baueri, M. lotae, M. tingrelaensis, M. shantipuri, M. rohitae, M. shetii, M. strelkovi and M. pfeiferri. An intercapsular process is absent in the species under study. But the presence of intercapsular process in M. strelkovi, M. pfeiferri, M. tingrelaensis, M. shetii, M. sangoricus, M. baueri, M. catostomi, M. minutus, M. lotae and a triangular notch in M. rohitae differentiates all of the above species from the present species. Ovoidal or tear-shaped spores of M. shantipuri with either equal or unequal polar capsules can also be differentiated from the spores of the present species.

In view of the above differences, the present species under study is proposed as new to the science and named as M. saugati sp. nov. through this communication.

M. szekeli sp. nov

Plasmodia

White, spherical to rounded, present on the internal wall of the stomach, 1–2 in number and measure 0.6–0.8 mm in diameter. 5–6 spores are present per plasmodium.

Spore description (Figs. 3a, b, 4a, b, 5b; Table 3) (measurements based on 4–8 spores in frontal view)

Fig. 3.

Fig. 3

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a Spore stained in Ziehl–Neelsen (valvular view). b Spore stained in iron–haematoxylin. Scale bar 0.005 mm

Fig. 4.

Fig. 4

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a Spore stained in Ziehl–Neelsen. b Spore stained in iron–haematoxylin

Table 3.

Measurements (in μm) and ratio of M. szekeli sp. nov.

Characters Range Mean values SD
LS 8.4–9.0 8.7 0.42
WS 3.9–4.3 4.1 0.28
LPC 3.9–4.9 4.4 0.70
WPC 1.0–2.0 1.5 0.70
Ratio: LS/WS 2.1
ICP Absent
NC 5–6
Parietal folds Absent
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The spores are histozoic, measure 8.7 × 4.1 μm, elongately ovoidal in valvular view having tapering, bluntly pointed anterior end and rounded posterior end. Two shell valves are thin, smooth, symmetrical and measure 0.3 μm in thickness. Parietal folds are absent. Polar capsules are two, equal, measure 4.4 × 1.5 μm, tubular with blunt anterior and rounded posterior ends. Polar capsules converge anteriorly but are placed at a distance posteriorly and occupies nearly half of the spore body cavity. Polar filaments form 5–6 coils and are arranged perpendicular to the polar capsule axis. An intercapsular process is absent. One capsulogenic nucleus is present measuring 1.3 μm in diameter. Sporoplasm is agranular and homogenous occupying whole of the extracapsular space behind the polar capsules. One sporoplasmic nucleus measuring 1.4 μm in diameter is present. An iodinophilous vacuole is present.

Taxonomic summary of M. szekeli sp. nov.

 

Type host Wallago attu vern. mulle
Type locality Harike wetland, Punjab, India
Type specimen Paratypes are spore stained in Ziehl–Neelsen and iron–haematoxylin, deposited in the museum of department of Zoology, Punjabi University, Patiala. Slide no. MH/ZN/01.03.2010 and MH/IH/01.03.2010.
Site of infection Internal wall of the stomach
Prevalence of infection 4% (1/25)
Etymology The species epithet szekeli has been given after the name of Dr. Csaba Szekely of Veterinary Medical Research Institute, Hungarian Academy of Sciences, Hungary.
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Discussion

The present species was compared with M. aligarhensis, M. mathuri, M. hosadurgensis, M. amieti, M. attui, M. bhadurius, M. trichogasteri, M. scatophagi, M. variformis, M. orissae and M. dhanachandi but differ from all of the above species in morphometric characteristics (Table 4).

Table 4.

Comparative description of M. szekeli sp. nov. with morphologically similar species (measurements in micrometer)

Species Host Site of infection Locality Spore Polar capsule
M. szekeli sp. nov. (present study) Wallago attu Internal wall of the stomach Harike wetland, Punjab (India) 8.7 × 4.1 4.4 × 1.5
M. aligarhensis Bhatt and Siddiqui 1964 Channa punctatus Accessory respiratory membrane, pharyngeal epithelium, fin West Bengal (India) 12.0–14.0 × 6.0–7.5 6.5–8.0 × 2.0–2.5 and 6.0–7.0 × 2.5
M. hosadurgensis Seenappa and Manohar 1981 Cirrhina mrigala Gill, muscles Karnataka (India) 10.5 × 6.25 5.37 × 2.3 and 3.3 × 1.43
M. mathuri Jayasri et al. 1981 Puntius sarana Gills Rajasthan (India) 8.7–23.5 × 5.1–10.1 2.7–11.9 × 1.8–4.6 and 2.7–7.8 × 1.8–4.6
M. amieti Fomena et al. 1985 Ctenopoma nanum Spleen, eye Cameroon 14.0 × 7.4 8.4 × 1.9
M. attui Sarkar 1985a W. attu Gut West Bengal (India) 13.89 × 8.53 5.92 × 3.0
M. bhadurius (Sarkar 1985a) Gupta and Khera 1988 W. attu Gall bladder West Bengal (India) 10.59 × 6.28 5.31 × 2.78
M. trichogasteri (Sarkar 1985b) Gupta and Khera 1988 Trichogaster fasciatus Gall bladder West Bengal (India) 15.55 × 9.35 10.1 × 3.32
M. scatophagi Haldar et al. 1996 Scatophagus argus Gills Orissa (India) 14.0 × 5.3 7.13 × 2.36
M. variformis Haldar et al. 1996 Mystus gulio Body muscles, gills Orissa (India) 15.2 × 5.65 9.67 × 2.84 and 8.6 × 2.28
M. orissae Haldar et al. 1996 C. mrigala Gills Orissa (India) 15.71 × 6.8 8.8 × 1.78 and 7.58 × 2.57
M. dhanachandi Bandyopadhyay et al. 2007 Channa orientalis Dorsal, ventral and caudal fins Manipur (India) 18.4 × 6.05 8.26 × 1.43
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The shape of the present species is elongately ovoidal with tapering, blunt anterior end and rounded posterior end. In this respect, the present species can be compared with M. aligarhensis, M. hosadurgensis and M. amieti in having spores with blunt anterior end as spores in rest of other species are pointed anteriorly. The species under study was also closely compared with M. dhanachandi on the basis of morphological similarities. But spores in M. dhanachandi are dumb-bell shaped, broader at the middle and tapering at both ends. Polar capsules are two, equal, tubular with blunt anterior but rounded posterior end and occupies nearly half of the spore body cavity in species under study. Unequal polar capsule in M. aligarhensis and M. hosadurgensis, very elongate polar capsules reaching two-third of the spore length in M. amieti and tear shaped polar capsules with anteriorly sharply pointed ends in M. dhanachandi differentiate them from the present species under consideration. In addition, both polar capsules in the present species converge anteriorly but are placed at a distance posteriorly in contrast to almost parallel polar capsules in M. dhanachandi.

In view of the above differences, the present species under study is proposed as new to the science and named as M. szekeli sp. nov. through this communication.

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