Five new earthworm species of the genera Amynthas and Metaphire (Megascolecidae: Oligochaeta) from Matsu, Taiwan Author Shen, Huei-Ping Zoology Division, Endemic Species Research Institute, Nantou, Taiwan; Author Chang, Chih-Han Department of Earth and Planetary Sciences, Johns Hopkins University, Baltimore, MD 21218, USA; Author Chih, Wen-Jay Department of Life Sciences and Institute of Zoology, National Taiwan University, Taipei, Taiwan text Journal of Natural History 2013 J. Nat. Hist. 2014-01-22 48 9 - 10 495 522 http://dx.doi.org/10.1080/00222933.2013.826742 journal article 10.1080/00222933.2013.826742 1464-5262 5199325 Metaphire matsuensis Shen , sp. nov. ( Figure 7 ) Type material Holotype . Clitellate (mature) specimen ( 84 mm in length, dissected) collected 28 June 2012 beside the Yanhsiu Fort (elevation 19 m ), Dongyin by H.P. Chen and W.J. Chih (TESRI-O-H-42) (voucher number: MTS 135). Table 3. A comparison of characters among Amynthas dongjuensis sp. nov. , Amynthas taiwumontis , Amynthas fornicatus , Amynthas penpuensis , Amynthas marenzelleri , Amynthas toriii , Amynthas baemsagolensis , Amynthas sinabunganus , Amynthas tertiadamae and Amynthas trinitatis .

Character	Amynthas dongjuensis	Amynthas taiwumontis	Amynthas fornicatus	Amynthas penpuensis	Amynthas marenzelleri	Amynthas toriii	Amynthas baemsagolensis	Amynthas sinabunganus	Amynthas tertiadamae	Amynthas trinitatis
sp. nov.	( Shen et al. 2013 )	( Gates 1935 ; Chen 1936 )	( Shen et al. 2003 )	( Cognetti 1906 ; Kobayashi 1938b )	( Ohfuchi 1941 )	( Hong and James 2001 )	( Michaelsen 1923 )	( Michaelsen 1934 )	( Beddard 1896 )
Locality	Matsu	Kinmen	Sichuan, China	Taiwan	Japan	Japan	Korea	Sumatra	Gulf of Siam	Trinidad
Body length (mm)	88–143	133–151	78–94	55–104	160–190	37–43	195–213	52	52	150
Clitellum width/ diameter (mm)	3.31–4.9	3.32–4.77	4–6	2.2–3.2	6–7	2.3–2.7	7.3–8.5	2–3	2.3–2.5	–
Segment number	107–122	131–142	90–105	62–104	130–138	77–83	109–132	104	120	100
First dorsal pore	12/13 or 13/14	11/12	11/12 or 12/13	5/6 or 6/7	12/13	12/13	12/13	13/14	–	11/12
Setal number
VII	31–41	74–97	43–50 (VI)	27–37	36–43	41–42	23	–	23 (VI)	–
VIII	–	–	46–52	–	–	43–45	–	50 (IX)	–	–
XII	–	–	42–52	–	45–49 (XIII)	–	–	50	40	–
XX	36–56	73–89	56	36–46	–	56–58	56	–	–	45 (XVII)
XXV	–	–	45–55	–	50–54 (XXIV)	–	–	53 (XXVI)	38	–

( Continued ) Table 3. (Continued).

Character	Amynthas	Amynthas	Amynthas	Amynthas	Amynthas	Amynthas	Amynthas	Amynthas	Amynthas	Amynthas
dongjuensis	taiwumontis	fornicatus	penpuensis	marenzelleri	toriii	baemsagolensis	sinabunganus	tertiadamae	trinitatis
sp. nov.	(Shen et al.	(Gates	(Shen	(Cognetti	(Ohfuchi	(Hong and	(Michaelsen	(Michaelsen	(Beddard
2013)	1935;	et al.	1906;	1941)	James 2001)	1923)	1934)	1896)
Chen	2003)	Kobayashi
1936)	1938b)
Between	5–11	16–22	9–14	8–11	8–10	7–8	12	–	–	20
male pores
Spermathecal	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9	5/6–8/9
pores
Male	Large,	Large,	Large, disk-shaped	Small,	Large, oval	Large,	Large, oval	Small, oval	Large,	Large,
porophore	round	round	or square	round	oblong	round	circular
Genital	Absent	Absent	Absent	Absent	Absent	Absent	Absent	Absent	Absent	Absent
papillae
Seminal	Large	Small	Small and elongate	Large	1st pair	2nd pair	Small	Large	Large	Large
vesicles	roundish,	larger
2nd pair
smaller
Prostate	Compact	Large in	–	Large in	Small in	Large in	XVII–XIX	Small in	Large in	Large in
glands	in	XVI–	XVII–	XVIII or	XVI–	XVIII	XVI–	XVII–
XVII–	XXIV	XX	XVII–	XX	XXII	XXI
XIX	XIX
Prostatic	Short, C-	Stout, C- or	–	Slender,	Short, C- or	Thin,	Thick, long	Long, bent	Stout, S-	Long,
Uducts	or S-	S-shaped	U-	S-shaped	straight	shaped	shaped
shaped	in XVIII	shaped
in	or	in
XVIII	XVIII–	XVII–
XIX	XVIII

Figure 7. Metaphire matsuensis sp. nov. (A) Right lateral view of spermathecal pore region of holotype (TESRI-O-H-42) (voucher number: MTS 135) (gp, genital papilla; sp, spermathecal porophore); (B) ventral view of preclitellar region of holotype; (C) left lateral view of male pore region of holotype (mp, male porophore; p, papilla); (D) ventral view of male pore region of the dissected paratype (TESRI-O-P-37) (voucher number: MTS 71); (E) dorsal view of right spermathecae of holotype (amp, ampulla; dv, diverticulum; ag, accessory gland); (F) dorsal view of left prostate gland of holotype. Scale bar 1 mm. Paratypes . Ten clitellates [one dissected (voucher number: MTS 71)] collected 31 May 2012 at Matsu Temple ( elevation 67 m ), Magang , Nangan by H.P. Chen and T . L. Ai (TESRI-O-P-37) . Other material Two clitellates collected 28 May 2012 along the Central Boulevard near the borders of Changshou and Chingshui villages (elevation 121 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–33) (voucher number of specimen used for DNA barcoding analyses: MTS 7); six clitellates collected 28 May 2012 at Ten Martyrs Monument (elevation 90 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–34); one clitellate collected 28 May 2012 at Shangyaoshan bus stop (elevation 99 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–35); two clitellates collected 28 May 2012 beside Beihai Tunnel (elevation 21 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–36); four clitellates collected 29 May 2012 in front of Banli Reservoir (elevation 33 m ), Beigan by H.P. Chen and T .L. Ai (coll. no. 2012–38) (voucher number of specimen used for DNA barcoding analyses: MTS 20); two clitellates collected 29 May 2012 from a roadside ditch at Chinbi Village (elevation 11 m ), Beigan by H.P. Chen and T .L. Ai (coll. no. 2012–39); 14 clitellates collected 29 May 2012 along Luoshan Nature Trail (elevation 66 m ), Beigan by H.P. Chen and T .L. Ai (coll. no. 2012–41) (voucher number of specimen used for DNA barcoding analyses: MTS 35); 13 clitellates collected 29 May 2012 at Junghsin Reservoir (elevation 54 m ), Beigan by H.P. Chen and T .L. Ai (coll. no. 2012–42) (voucher number of specimen used for DNA barcoding analyses: MTS 37); one clitellate collected 29 May 2012 in front of Matsu Broadcasting Station (elevation 205 m ), Beigan by H.P. Chen and T .L. Ai (coll. no. 2012–43); six clitellates collected 31 May 2012 from a roadside ditch near the weather station (elevation 48 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–54); 12 clitellates collected 31 May 2012 along the road facing Huangguan Channel (elevation 48 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–56) (voucher number of specimen used for DNA barcoding analyses: MTS 77); three clitellates collected 31 May 2012 in front of Tunnel 88 (elevation 62 m ), Nangan by H.P. Chen and T .L. Ai (coll. no. 2012–57); eight clitellates collected 25 June 2012 from roadside ditches (elevation 39 m ), Hanlinjiao, Nangan by H.P. Chen and W.J. Chih (coll. no. 2012–58) (voucher numbers of specimens used for DNA barcoding analyses: MTS 81, 82, 83); four clitellates collected 25 June 2012 at Stronghold No. 12 (elevation 59 m ), Nangan by H.P. Chen and W.J. Chih (coll. no. 2012–60); six clitellates collected 26 June 2012 from Mount Nigu (elevation 59 m ), Beigan by H.P. Chen and W.J. Chih (coll. no. 2012–64) (voucher number of specimen used for DNA barcoding analyses: MTS 110); one clitellate collected 27 June 2012 above the tidal zone (elevation 8 m ) near Dapu Stone Inscription, Dongju by H.P. Chen and W.J. Chih (coll. no. 2012–68); two clitellates collected 28 June 2012 at Stronghold No. 33 (elevation 61 m ), Hsiyin by H.P. Chen and W.J. Chih (coll. no. 2012–72); five clitellates collected 28 June 2012 from Chingshuiao (elevation 15 m ), Hsiyin by H.P. Chen and W.J. Chih (coll. no. 2012–74); seven clitellates collected 28 June 2012 near Northernmost Frontier (elevation 79 m ), Hsiyin by H.P. Chen and W.J. Chih (coll. no. 2012–75) (voucher numbers of specimens used for DNA barcoding analyses: MTS 129, 130); five clitellates collected 28 June 2012 from a roadside ditch near Stronghold No. 26 (elevation 19 m ), Hsiyin by H.P. Chen and W.J. Chih (coll. no. 2012–76); three clitellates collected 28 June 2012 in front of Beihai Tunnel (elevation 24 m ), Dongyin by H.P. Chen and W.J. Chih (coll. no. 2012–77) (voucher numbers: MTS 131, 132, 133); three clitellates collected 28 June 2012 beside the Yanhsiu Fort (elevation 19 m ), Dongyin by H.P. Chen and W.J. Chih (coll. no. 2012–78) (voucher numbers of specimens used for DNA barcoding analyses: MTS 134, 136); one clitellate collected 28 August 2012 from a roadside ditch in Nioubei Ridge (elevation 138 m ), Nangan by H.P. Chen and H.H. Huang (coll. no. 2012–116). Diagnosis Small to medium; length (clitellates) 58–135 mm . Segments numbering 95–100. Setae 50–68 in VII, 53–74 in XX, 11–18 between male pores. First dorsal pore in 12/13. Spermathecal pores dorsolateral, three pairs in 5/6–7/8, 0.34–0.37 body circumference dorsally apart. Male pores 0.28–0.38 body circumference ventrally apart in XVIII, each pore surrounded by 0–10 small papillae on top of a cone-shaped everted copulatory chamber, the secondary aperture of each copulatory chamber as a longitudinal slit when the chamber invaginated, wrinkled around its margin. Spermathecae three pairs in VI–VIII. Seminal vesicles large, two pairs in XI and XII. Prostate glands large in XVI–XXI or XVII–XXI, wrinkled and lobed. Prostatic duct stout, U-shaped in XVIII. Accessory glands stalked, mushroom-like or irregularshaped, corresponding to external genital papillae. Description External characters. Total length (clitellates) 58–135 mm . Weight 0.26–2.26 g . Segments numbering 95–100. Clitellum XIV–XVI, setae and dorsal pores absent, length 3.14–6.31 mm and width 2.52–5.73 mm . Prostomium epilobous. Setae minute, number 50–68 in VII, 53–74 in XX, and 11–18 between male pores in XVIII. First dorsal pore in 12/13. Spermathecal pores lip-like, dorsolateral, three pairs in intersegmental furrows of 5/6–7/8 ( Figure 7A ), distance between paired pores 0.34–0.37 body circumference dorsally apart. Each pore surrounded by two tiny papillae: one anterior and the other posterior. Occasionally one or two papillae medial to each pore ( Figure 7A ). Genital papillae absent, one to three median, or one to three lateral on both left and right sides in presetal VIII ( Figure 7B ). Female pore single, mid-ventral in XIV. Male pores paired in XVIII, 0.28–0.38 body circumference ventrally apart. In the case of protrusion, each pore surrounded by 0–10 small papillae on top of a coneshaped everted copulatory chamber ( Figure 7C ). The secondary aperture of each copulatory chamber as a longitudinal slit when the chamber invaginated, wrinkled around its margin ( Figure 7D ). The whole male area surrounded by several concentric skin folds. Preserved specimens brown to greyish brown in colour with brown clitellum. Internal characters. Septa 5/6–7/8 and 10/11–13/14 thick, 8/9 membranous, 9/10 absent. Nephridial tufts on anterior faces of 5/6/7. Gizzard large in VIII–X. Intestine enlarged from XV. Intestinal caeca paired in XXVII, extending anteriorly to XXIV–XXV, each simple, wrinkled, distal end either straight or bent. Oesophageal hearts in X–XIII. Spermathecae three pairs in VI–VIII, each with a peach-shaped or elongated ovalshaped ampulla 1.79–3.07 mm long and 1.2–3.06 mm wide, and a stout spermathecal stalk 0.63–1.0 mm in length. Diverticulum slender and straight at proximal part, coiled and enlarged at distal part ( Figure 7E ). Accessory glands stalked, mushroomlike, each corresponding to external genital papilla, about 1.0 mm in total length. Holandry: testes large, two pairs in ventrally joined sacs in X and XI. Seminal vesicles large, finely folliculated, two pairs in XI and XII, occupying the full compartment, usually anterior pair larger, without a dorsal lobe. Prostate glands large in XVI–XXI or XVII–XXI, wrinkled and lobed. Prostatic duct stout, U-shaped in XVIII. Accessory glands stalked, mushroom-like or irregular-shaped, 0.46–1.08 mm in total length, corresponding to external genital papillae ( Figure 7F ). DNA barcodes GenBank accession numbers KC897033 (MTS 135, holotype ), KC897038 (MTS 71, dissected paratype ), KC897029 , KC897030 , KC897025 , KC897039 , KC897028 , KC897031 , KC897027 , KC897037 , KC897032 , KC897040 , KC897041 , KC897034 , KC897026 , KC897024 , KC897036 and KC897035 (MTS 7, 20, 35, 37, 77, 81, 82, 83, 110, 129, 130, 131, 132, 133, 134 and 136, respectively) . Etymology The name matsuensis is given to this species with reference to its type locality of the islands of Matsu . Remarks Metaphire matsuensis sp. nov. is common on the islands of Nangan, Beigan, Dongyin and Hsiyin. However, only one specimen was collected on Dongju and none was found on Hsiju. It is easily distinguishable from all other Metaphire species with three pairs of spermathecal pores in intersegmental furrows of 5/6–7/8 listed in Sims and Easton (1972) simply by the male pore structure. Instead, its male pore structure is morphologically fairly similar to Metaphire guillelmi ( Michaelsen, 1895 ) from Hupei, China and Metaphire bifoliolare Tan and Zhong, 1987 from Hunan , China . Metaphire guillelmi is a sexthecate earthworm with three pairs of ventrolateral spermathecal pores in intersegmental furrows of 6/7–8/9 ( Michaelsen 1895 ). The structure of its spermatheca is morphologically fairly similar to M. matsuensis . Metaphire bifoliolare is also sexthecate with three pairs of intrasegmental spermathecal pores in posterior V–VII ( Tan and Zhong 1987 ). It has slender, straight diverticulum with a sausage-shaped seminal chamber, which is different from the coiled diverticulum of M. matsuensis . In addition, the internal and external morphological characters of M. matsuensis are also similar to the sexthecate Metaphire thecodorsata ( Chen, 1933 ) from Jiangxi , China , but the spermathecal pores of the latter are large transverse slits close to the dorsomedian line in posterior V–VII ( Chen 1933 ). All the above three species, M. guillelmi , M. bifoliolare and M. thecodorsata , are distributed along the lower Yangtze Valley, central China . The male pore structure of M. matsuensis is also similar to that of Metaphire soulensis (Kobayashi, 1938) from Korea . However, M. soulensis is quadrithecate with two pairs of ventrolateral spermathecal pores in intersegmental furrows of 6/7/8. The characteristics of complex caeca consisting of five or six finger-shaped processes, absence of prostate glands and prostatic ducts, and vestigial spermathecae without diverticulum of M. soulensis ( Kobayashi 1938a ) are distinctly different from those of M. matsuensis .