Coynema poeyi ( Coy, García & Álvarez, 1993 ), Morffe & Garcia, 2011

Morffe, Jans, García, Nayla, Breugelmans, Karin, Adams, Byron J. & Hasegawa, Koichi, 2018, Morphological and molecular characterization of Coynema poeyi (Coy, García & Álvarez, 1993) (Oxyuridomorpha: Hystrignathidae) from Antillanax pertyi (Kaup, 1869) (Coleoptera: Passalidae) from Cuba and new locality records for the species, Zootaxa 4497 (1), pp. 29-40 : 31-38

Coynema poeyi ( Coy, García & Álvarez, 1993)

Fig. 1 A–H , Fig. 2 A–G , Fig. 3 A–F , Fig. 4 A–F

Glaber poeyi Coy, García & Álvarez, 1993: 57 –59, fig. 3 A–E

Coynema poeyi ( Coy, García & Álvarez, 1993) Morffe & García, 2011: 9 –12, fig. 1 A–H, 2 A–C, 3 A–H, new combination

Material examined. Vouchers: 10♀♀, Cuba, Pinar del Río province, San Diego de los Baños, Las Yeguas ; in

Antillanax pertyi ; 31/V/2014; S. Yong coll.; CZACC 11.7061–11.7070. 9♂♂, same data as the latter; CZACC 11.7071–11.7079 .

Vouchers: 7♀♀, Cuba, Mayabeque province, Lomas del Cheche ; in Antillanax pertyi ; VI/2013; J. Morffe, N. García coll.; CZACC 11.7080–11.7086 . 7♂♂, same data as the latter; CZACC 11.7087–11.7093 . 4♀♀, same data as the latter; MNHNSD 05.0012–05.0015 . 4♂♂, same data as the latter; MNHNSD 05.0016–05.019 .

Vouchers: 9♀♀, Cuba, Sancti Spíritus province , Trinidad, Topes de Collantes, path to the River Caburní ; in Antillanax pertyi ; 12/X/2014; J. Morffe, N. García coll.; CZACC 11.7094–11.7102 . 6♂♂, same data as the latter; CZACC 11.7103–11.7108 .

Vouchers: 4♀♀, Cuba, Guantánamo province , El Salvador, Limonar ; in Antillanax pertyi ; VI/2013; J. Morffe, N. García, M. Olcha coll.; CZACC 11.7109–11.7112 . 2♂♂, same data as the latter; CZACC 11.7113–11.7114 .

Redescription. General. CerVical cuticle unarmed. Sub-cuticular Striae preSent. Head Set-off from the body by a deep grooVe, bearing eight paired, elliptical and flat cephalic papillae. The papillae of each pair are oblique, With their inner tipS conVerging and almoSt contacting each other, forming a V-like arrangement. AmphidS lateral, aS reniform poreS. Buccal aperture circular. Stoma Short, Slightly conoid and Surrounded by an oeSophageal collar. OeSophaguS conSiStS of a muScular, Sub-cylindrical procorpuS, itS baSe abruptly dilated at itS joint With the Short iSthmuS. BaSal bulb pyriform, ValVe plate Well-deVeloped. InteStine Simple, Sub-rectilinear, anterior portion notably dilated, forming a Sac-like Structure Slightly longer than oeSophaguS. At thiS part, the external Surface of inteStine almoSt toucheS the body Wall. NerVe ring encircling procorpuS at about 60% of itS length. Excretory pore Ventral, Situated at about half of a body Width poSterior to baSal bulb.

Female. Body robuSt, markedly Spindle-Shape, WidenS gradually poSterior to head, reaching itS maximum Width at leVel of the anterior portion of the inteStine, then narroWing gradually toWardS tail. CerVical cuticle unarmed and finely annulated (annuli ca. 2 µm). In addition to the more marked ridgeS that diVide the annuli, each one iS Sub-diVided by a median, ShalloW grooVe. Lateral alae extend from the end of the procorpuS or the beginning of itS baSal dilation to about half of a body Width before the leVel of VulVa. FirSt cephalic annule Slightly dilated, Short, conoid and truncate, almoSt aS long aS the head. Excretory pore Ventral located at ca. half of the body-Width poSterior the baSal bulb. Rectum Short, anuS not prominent. Excretory pore Surrounded by a prominent cuticular ring. VulVa a Ventro-median tranSVerSe Slit, located near the mid-body or Slightly diSplaced to the poSterior half of body. Anterior lip of VulVa prominent. Vagina vera muScular, thin-Walled, forWardly directed. Vagina uterina reflexed tWice, firSt poSteriorly directed and then anteriorly directed before joint to the common uteruS. Genital tract didelphic-amphidelphic. Both oVarieS reflexed, diStal flexureS Very Short, ca. one third of the body Width. Anterior oVary reflexed juSt poSterior to the Sac-like Structure of the inteStine. PoSterior oVary reflexed at ca. tWo body-WidthS before the leVel of the anuS. OocyteS in Single roWS. EggS comparatiVely large, markedly oVoid in Shape, Smooth-Shelled. GraVid femaleS With 1–10 eggS at a time in the uteri, more frequently 4–5. Tail long, filiform and Subulate, ending in a Sharp tip.

Male. Body Smaller and leSS robuSt than femaleS, poSterior end Ventrally curVed. CerVical cuticle unarmed and finely annulated (annuli ca. 1.5 µm). In addition to the more marked ridgeS that diVide the annuli, each one iS SubdiVided by a median, ShalloW ridge. Lateral alae narroW, from ca. the baSe of the procorpuS to ca. 20 µm before the leVel of the cloaca. Without eVident firSt cephalic annule. Monorchic. TeStiS Ventral, outStretched, commencing juSt poSterior to the Sac-like Structure of the inteStine. Vas deferens With three diStinguiShable regionS: an anterior region (occupying ca. the firSt third of the teStiS) With granular content and SWollen in itS joint With a median region With large, rounded cellS, and a poSterior region that diminiSheS itS diameter through the cloaca. Spicule abSent. Tail Very Short, conoid, itS tip truncate. DorSal cuticle of the tail end thickened and Smooth, from the Sub-dorSal precloacal papillae to the tail tip. Eight copulatory papillae preSent, four pre-cloacal and four poSt-cloacal. Pre-cloacal papillae conSiSt of tWo pairS of large, mammiliform papillae: one Sub-Ventral, located a Short diStance before the cloaca (ca. 10 µm) and the other Sub-dorSal, near the leVel of the cloaca. TWo pairS of minute lateral poSt-cloacal papillae: one near the tail tip (ca. 3 µm from it) and the other at the tail tip. PhaSmidS Sub-dorSal, located at ca. 10 µm from the tail tip.

Remarks. Morffe & García (2011) deScribed C. poeyi baSed only on light microScopy. HoWeVer, the preSent SEM StudieS ShoW featureS not conSidered in the preViouS Work. The authorS indicated that C. poeyi preSentS eight paired papillae in the head, Without detailS on their Shape and arrangement. By meanS of SEM theSe papillae can be obSerVed in both SexeS, each pair aS elliptical and flat, being oblique and forming a V-like arrangement With their inner tipS conVerging and almoSt contacting each other. Such papillae are unuSual, Since moSt of the SpecieS Studied by SEM or by en face VieWS under light microScopy of the cephalic end preSent rounded papillae (i.e. Hunt 1981, 1982; Morffe & García 2013a, b; Morffe et al. 2015; Van Waerebeke & Remillet 1982). In contraSt, the genera Longior TraVaSSoS & KloSS, 1958 and Papillabrum KloSS, 1962 poSSeSS digitiform papillae ( KloSS 1962; Morffe et al. 2018).

The preSence of cerVical annuli WaS mentioned by Morffe & García (2011) only in femaleS. Here, We confirm that maleS alSo preSent an annulated cerVical cuticle. In addition, We obSerVed that each annule iS Sub-diVided by a median grooVe.

In the deScription of the maleS, Morffe & García (2011) did not mention the preSence of lateral alae in thiS Sex. HoWeVer, in the current Study We obSerVed narroW lateral alae extending from ca. the baSe of the procorpuS to ca. 20 µm before the leVel of the cloaca.

The StructureS of the tail end of thelaStomatoid maleS are hard to obSerVe under light microScopy, eSpecially in Small SpecimenS. Morffe & García (2011) obSerVed a Y-like thickening of the dorSal cuticle of the tail, Which iS Smooth according to SEM imageS. ThiS pointS to What We interpret to be an artifact during the obSerVation of the SpecimenS under Such light microScopy. Regarding the copulatory papillae, Morffe & García (2011) only obSerVed “A Single, median, large, mammiform pre-cloacal papilla…” and “A pair of Small, Sub-dorSal, pre-cloacal papillae Situated before the dorSal cuticular thickening.”. By SEM obSerVationS Such Ventromedian papilla actually correSpondS to the Sub-Ventral pair of pre-cloacal papillae. The current Work alSo reVealed tWo additional pairS of poSt-cloacal papillae, lateral and minute near the tail tip and at the tail tip, reSpectiVely.

So far, C. poeyi haS been recorded moStly for localitieS from WeStern Cuba: El Salón, Sierra del RoSario, ArtemiSa proVince (itS type locality), El Mulo, Sierra del RoSario, ArtemiSa proVince and La Jaula, Mayabeque proVince. La Melba, Holguín proVince, in the mountain range of Nipe-Sagua-Baracoa, EaStern Cuba iS the other knoWn record for the SpecieS ( Coy et al. 1993; Morffe & García 2011).

Of the preSent neW recordS for C. poeyi , LoS PalacioS, Pinar del Río proVince and Limonar, Guantánamo proVince are the WeSternmoSt and the eaSternmoSt for the country, reSpectiVely. LomaS del Cheche in the AlturaS Habana-MatanzaS mountain range, Mayabeque proVince, iS cloSe to the preViouS record of La Jaula. On the other hand, Caburní, Sancti SpírituS proVince iS the firSt record for thiS SpecieS in Central Cuba. The indiVidualS from the aforementioned neW localitieS coincide morphologically and morphometrically ( TableS 1 and 2) With the preViouS recordS of the SpecieS.

DNA studies. TWo partial SequenceS (ca. 790 bp) of the D2-D3 region of the 28S LSU rDNA Were obtained from femaleS of C. poeyi . No intra-Specific differenceS Were recorded betWeen the Studied populationS.

The ML and BI treeS ShoWed identical topology, thuS only the ML tree iS ShoWn ( Fig. 5 ). The phylogenetic analySeS of the D2-D3 28S LSU rDNA ShoW that C. poeyi formS a monophyletic clade With the other aVailable SequenceS of HyStrignathidae : Hystrignathus Sp. (from Vietnam) and tWo Cuban SpecieS of Longior . In both methodS C. poeyi iS located baSal on Such clade.

The poSition of C. poeyi in the clade can be morphologically explained Since thiS monotypic genuS preSentS a feature unique for the family, namely the Sac-like Structure of the inteStine, conSiSting of a conSpicuouS dilation of itS anterior region, longer than the oeSophaguS and With the external Surface of the inteStine almoSt touching the body Wall. In the other hyStrignathid genera the anterior part of the inteStine can be more or leSS dilated, but thiS SWollen area iS not longer than the oeSophaguS length and the Surface of the inteStine iS far from touching the body Wall. MoreoVer, the oeSophaguS of Coynema, With the Sub-cylindrical procorpuS and SWollen baSe iS preSent only in the genera Glaber TraVaSSoS & KloSS, 1958 and Vulcanonema TraVaSSoS & KloSS, 1958 ( TraVaSSoS & KloSS 1958). Of the genera examined in our phylogenetic analySiS, Longior preSentS an elongated, Sub-cylindrical procorpuS WhereaS Hystrignathus Leidy, 1850 haS a claVate procorpuS (AdamSon & Van Waerebeke 1992).

The cephalic papillae of Coynema , elliptical and in a V-like pattern are alSo not frequent in the group. AS mentioned aboVe, moSt of the hyStrignathidS preSent rounded papillae and a feW genera haVe digitiform papillae. In the caSe of Hystrignathus and Longior , they preSent rounded and digitiform papillae, reSpectiVely ( Hunt 1982; Morffe et al. 2015; Morffe et al. 2018).

Figures

FIGURE 1. Coynema poeyi (Coy, García & Álvarez, 1993). Female. A. Oesophageal region, lateral view. B. Tail, lateral view. C. Cephalic end, optical section. D. Cephalic end, external view (reconstructed from SEM images). E. Genital tract, lateral view. F. Uagina vera and vagina uterina, lateral view. G. Egg. H. Habitus, lateral view.

FIGURE 2. Coynema poeyi (Coy, García & Álvarez, 1993). Male. A. Oesophageal region, lateral view. B. Cephalic end, optical section. C. Cephalic end, external view (reconstructed from SEM images). D. Cephalic end, en face view (reconstructed from SEM images). E. Tail region, lateral view. F. Tail region, dorsolateral view (reconstructed from SEM images). G. Habitus, lateral view.

FIGURE 3. Coynema poeyi (Coy, García & Álvarez, 1993). Female. SEM images. A. Habitus, lateral view (white arrowhead point the beginning and end of the lateral alae, black arrowhead points the vulva). B. Anterior half of body, showing the beginning and end of lateral ala (pointed by arrowheads). C. Anterior end. D. Cephalic end, en face view (arrowheads point the amphids). E. Detail of the cephalic end, en face view (arrowheads point the amphids). F. Detail of the excretory pore. Scale bars: A. 500 µm. B. 250 µm. C. 15 µm. D, E. 10 µm. F. 5 µm.

FIGURE 4. Coynema poeyi (Coy, García & Álvarez, 1993). Male. SEM images. A. Oesophageal region. B. Cervical region, lateral view. C. Cephalic end, en face view. D. Posterior end, ventrolateral view. E. Detail of the tail end, dorsolateral view (arrowheads point the phasmids). F. Detail of the tail end, frontal view. Scale bars: A. 50 µm. B, D, E. 10 µm. C, F. 5 µm.

FIGURE 5. Maximum likelihood (ML) tree inferred from D2-D3 28S LSU rDNA for several species of the superfamily Thelastomatoidea (Nematoda: Oxyuridomorpha). Cameronia multiovata (Thelastomatidae) and three species of Travassosinema (Travassosinematidae) were used as outgroup taxa. Values at the nodes correspond to bootstrap resampling/ posterior probability.