Indopolystoma n. gen. (Monogenea, Polystomatidae) with the description of three new species and reassignment of eight known Polystoma species from Asian frogs (Anura, Rhacophoridae) Author Chaabane, Amira Unit for Environmental Sciences and Management, North-West University, Potchefstroom Campus, Private Bag X 6001, Author Verneau, Olivier Unit for Environmental Sciences and Management, North-West University, Potchefstroom Campus, Private Bag X 6001, & University of Perpignan Via Domitia, Centre de Formation et de Recherche sur les Environnements Méditerranéens, & CNRS, Centre de Formation et de Recherche sur les Environnements Méditerranéens, UMR 5110, 66860 Perpignan, France Author Preez, Louis Du Unit for Environmental Sciences and Management, North-West University, Potchefstroom Campus, Private Bag X 6001, & South African Institute for Aquatic Biodiversity, Private Bag 1015, 6140 Grahamstown, South Africa text Parasite 2019 Paris, France 2019-11-20 26 67 1 18 http://dx.doi.org/10.1051/parasite/2019067 journal article 10.1051/parasite/2019067 1776-1042 PMC6865761 31746733 12524337 71763E07-E6BC-4FB7-94B0-03132D46FFAF Indopolystoma elongatum n. gen. n. sp. ( Figs. 5 and 6 ; Table 4 ) urn:lsid:zoobank.org:act: EDFE29A4-9B26-4A25-A92ABB3CF106D4EC Synonym: Polystoma sp. of Badets et al. [ 2 ] and Héritier et al. [ 22 ]. Type-host: Rhacophorus arboreus (Okada & Kawano) . Now Zhangixalus arboreus (Okada and Kawano) [ 21 , 27 ]. Site: Bladder. Type-locality: Upstream of Kunigami-gun, city of Nago, Okinawa prefecture , Japan . Other localities: Sado Island, Niigata prefecture , Japan . Collector: Professor Hideo Hasegawa , Department of Biology, Oita Medical University, Hasama, Oita 879-5593, Japan . Type-specimens: Holotype ( MNHN HEL1184 ) and 2 paratypes ( MNHN HEL1185 – HEL1186 ) deposited in the Parasite Collection, National Museum of Natural History, Paris, France . Table 4. Body measurements of Indopolystoma spp.

Parasite species	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma	Indopolystoma
viridi n. sp.	elongatum n. sp.	parvum n. sp.	carvirostris	hakgalense	indicum	leucomystax	mutus n. comb.	pingbianensis	rhacophori	zuoi
n. comb.	n. comb.	n. comb.	n. comb.	n. comb.	n. comb.	n. comb.
Host species	Zhangixalus	Zhangixalus arboreus Zhangixalus		Kurixalus bisacculus	Taruga	Rhacophorus	Polypedates	Polypedates	Zhangixalus	Zhangixalus	Feihyla
viridis	omeimontis	or Kurixalus	eques	nigropalmatus	leucomystax	mutus	dugritei	arboreus	palpebralis
verrucosus
Site	Bladder	Bladder	Bladder	Bladder	Bladder	Bladder	Bladder	Bladder	Bladder	Bladder	Bladder
Locality	Japan	Japan	China	China	Sri Lanka	India	China	China	China	Japan	China
Number of mature specimens	15	3	2	10	12	8	5	2	6–7	–	3
Body length	8550	14,791	4714	5070	(3870–9230)	12,556	7568	7338	9428	(4500–6000)	2716 (1624–3533)
(5532–11,907)	(12,847–14,878)	(4536–4891)	(4030–7790)	(6732–16,614)	(6665–8514)	(6875–7800)	(6000–12,000)
Greatest width	2112 (1534–2859)	3182 (3170–3270)	1916 (1701–2130)	1130 (810–1400)	(890–1790)	4164 (2222–5641)	2732 (2542–3311)	3063 (2550–3575) 1990 (1640–2390) (1000–1900)			1280 (1202–1904)
Width at vagina	1481 (1175–1800)	1967 (1902–2031)	1402 (1279–1525)	830	–	2057 (1197–2622)	–	–	1572	–	–
Haptor length	773 (526–1354)	916 (734–1098)	667 (584–749)	680 (500–750)	(635–794)	1298 (977–1710)	824 (753–989)	913 (875–950)	869 (424–1253)	(800–1100)	701 (632–924)
Haptor width	1184 (588–1592)	1661 (1512–1810)	1258 (1022–1494)	890 (640–1500)	(1016–1509)	2104 (1584–2466)	1693 (1355–2040)	1650 (1425–1875) 784 (415–1374)		(1100–1600)	1063 (831–1317)
False oral sucker	394 (266–465)	551 (528–573)	395 (358–431)	–	(189–402)	466 (236–574)	392 (194–424)	388 (275–500)	–	–	–
width
Pharynx length	210 (124–268)	237 (227–247)	191 (187–194)	–	–	291 (164–395)	252 (219–286)	245 (210–280)	253 (218–328)	(188–240)	164
Pharynx width	209 (164–244)	259 (256–268)	190 (174–205)	–	(135–197)	249 (164–349)	216 (194–246)	235 (200–270)	229 (189–252)	(188–220)	151
Ovary length	775 (577–925)	1072 (1062–1108)	545 (483–606)	497	(492–874)	1049 (504–1368)	608 (451–683)	588 (550–625)	711.5 (453–927)	(420–630)	369 (329–400)
Ovary width	401 (286–536)	520 (514–563)	269 (231–306)	390	(207–349)	499 (288–630)	259 (191–294)	400 (375–425)	–	(210–340)	–
Length of eggs	241 (191–268)	240 (224–256)	221 (219–222)	–	–	205 (190–214)	–	–	307	(238–300)	–
Width of eggs	125 (74–165)	118 (106–130)	103 (93–113)	–	–	132 (125–140)	–	–	165	(140–163)	–
Number of	1	1	1	0	–	up to 40	0	0	1–8	Usually 1 but	0
intra-uterine eggs	sometimes
more than 10
Genital bulb width	105 (67–124)	139 (129–211)	83 (75–90)	–	–	–	(69–76)	–	–	–	–
Number of genital	8–9	8	8	8	–	8	8	9	–	8	8
spines
Length of genital	40 (27–49)	41 (38–44)	17 (16–18)	–	–	39 (38–40)	–	39 (38–40)	–	42	–
spine
Haptoral sucker	333 (242–423)	420 (380–459)	335 (281–389)	259 (212–339)	(250–320)	353 (264–465)	289 (207–356)	310 (270–350)	434 (349–491)	(320–400)	251 (209–311)
width
Hamulus handle length	313 (276–373)	407 (303–419)	326 (311–340)	285 (257–326)	(380–440)	320 (178–414)	332 (260–410)	340 (330–350)	408 (332–441)	(350–420)	240 (185–307)
Hamulus guard length	–	–	–	249 (208–306)	–	–	–	290 (280–300)	382 (340–461)	–	205 (173–265)
Hamulus hook length	66 (48–74)	78.5 (72–85)	52 (39–64)	–	–	–	–	40 (38–43)	–	–	–
Marginal hooklet length	–	–	–	(16–18) only on suckers	(20.5–36.9) C1–C2	–	–	–	–	(24–42)	–
Marginal hooklet	40 (31–44)	36	32	–	–	–	–	–	–	(38–42)	–
C1 length
Marginal hooklet	21.5 (16–31)	23 (18–32)	(19–20)	–	–	–	–	–	–	(24–27)	–
C2–C8 length
Haptoral length/Body length	0.09 (0.05–0.17)	0.06 (0.05–0.07)	0.14 (0.12–0.165)	0.13	0.11	0.10	0.11	0.12	0.09	0.18	0.26
Number of anastomoses	0	>1	1	>1	>1	1–2	>1	>1	0–1	>1	3–4

A. Chaabane et al.: Parasite 2019, 26 , 67 Note. To the exception of the newly described species, body measurements for all other species were extracted or estimated from Crusz and Ching [ 12 ], Diengdoh and Tandon [ 13 ], Fan et al. [ 19 , 20 ], Meng et al. [ 33 ], Shen et al. [ 40 ], Yamaguti [ 53 ] and Zhang and Long [ 54 ]. “–” means data missing. 7 Figure 2. Indopolystoma viridi n. gen. n. sp. from Zhangixalus viridis . Dorsal view of holotype. (A) A drawing; (B) A photograph. Scale bar: 1 mm. Etymology: The species name elongatum refers to its general body shape, which is elongate. Description Description based on three flattened specimens stained in carmine and mounted in Canada balsam. Body elongate, tapered anteriorly, total length 14,791 ( 12,847 –14,878 ), including haptor; greatest width 3182 (3170–3270); width at vagina 1967 (1902–2031). Tegument smooth. Haptor sub-rectangular, 916 (734–1098) long, 1661 (1512–1810) wide. Haptor/total body length ratio 0.06 (0.05–0.07). Suckers 420 (380–459) in diameter. Hamuli 407 (303–419) long; with hook 78.5 (72–85) long ( Fig. 6A–A ’’’). Marginal hooklet C1 36 long; C2–C8 23 (18–32) long ( Fig. 6B–B ’’). Mouth ventral, sub-terminal and surrounded by false oral sucker; false oral sucker 551 (528–573) wide. Pharynx pyriform, 237 (227–247) long, 259 (256–268) wide. Oesophagus not visible. Intestine bifurcate with medial diverticula highly branched giving rise to prehaptoral anastomoses (up to two). Testis not visible hidden by the digestive tract and vitellaria. Seminal vesicle prominent and packed with sperm. Genital bulb slightly sclerotized, medio-ventral, 139 (129–211) in diameter, with eight sclerotized genital spines; genital spines 41 (38–44) long. Ovary prominent, submedian and packed with oocytes; ovary 1072 (1062–1108) long, 520 (514–563) wide. Ootype well developed. Genito-intestinal canal present on same side of body as ovary, joining intestinal caecum posterior to ovary. Uterus confined to area anterior to ovary holding one egg ; egg 240 (224–256) long, 118 (106–130) wide. No intrauterine development of eggs observed ( Fig. 5 ). A. Chaabane et al.: Parasite 2019, 26 , 67 9 Figure 3. Indopolystoma viridi n. gen. n. sp. from Zhangixalus viridis . Hamuli from holotype and paratypes. (A–A’) Holotype; (B–H) Paratypes. Scale bar: 100 µm. Differential diagnosis Indopolystoma elongatum is well characterized by its body size and shape. This species is much bigger and more elongated (body length 14,791 µm) than any other species of Indopolystoma , though there is an overlap of size values with I. indicum . Indopolystoma elongatum can be easily distinguished from the later by the number of intrauterine eggs. None of the specimens of I. elongatum have more than a single egg in utero while I. indicum has as many as 40. Remarks: Zhangixalus arboreus hosts two polystomes, namely I. elongatum and I. rhacophori (see below), which is uncommon within anuran polystomes. However, Z. arboreus and Z. schlegelii occur sympatrically in Japan [ 1 ]. The possibility of a misidentification can thus not be excluded especially since molecular evidence on host identity is currently not available. We consider for now that both I. elongatum and I. rhacophori are separate species primarily on the basis of body length and haptor/total body length ratio (0.06 for I. elongatum vs. 0.18 for I. rhacophori ).