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A NEW CUCULLANID SPECIES (NEMATODA) FROM AGENEIOSUS UCAYALENSIS CASTELNAU, 1855 (PISCES: AUCHENIPTERIDAE) FROM PAR , BRAZIL

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A NEW CUCULLANID SPECIES (NEMATODA) FROM AGENEIOSUS UCAYALENSIS CASTELNAU, 1855 (PISCES: AUCHENIPTERIDAE) FROM PAR , BRAZIL
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  A NEW CUCULLANID SPECIES (NEMATODA) FROM  AGENEIOSUS UCAYALENSIS  CASTELNAU, 1855 (PISCES: AUCHENIPTERIDAE) FROM PARA´ , BRAZIL Elane G. Giese, Adriano P. Furtado, Reinalda M. Lanfredi, and Jeannie N. Santos * Laborato´rio de Biologia Celular e Helmintologia ‘‘Profa. Dra. Reinalda Marisa Lanfredi’’, Instituto de Cieˆncias Biolo´gicas, UFPA, Rua AugustoCorreˆa, s/n. CEP 66075-110, Para´, Bele´m, Brazil.  e-mail: jeannie@ufpa.br  ABSTRACT :  Cucullanus ageneiosus  n. sp. (Nematoda: Cucullanidae) is described from the intestine of the  Ageneiosus ucayalensis , fromthe Guajara´ Bay, Bele´m, Para´, Brazil. The new species is characterized mainly by arrangement of male caudal papillae, and the positionof lateral phasmids immediately posterior of 10th papillae, markedly short and unequal spicules, a precloacal sucker, and the presenceof an unpaired median papilla just of the anterior cloacal lip. Cucullanus  Mu¨ller, 1777 includes approximately 100 species, allparasites of fishes around the world (Timi and Lanfranchi, 2006).After the description of   Cucullanus cirratus  Mu¨ller, 1777, the firstspecies described to the Neotropical region, 26 additionalspecies of  Cucullanus have beendescribed fromthe same area,ofwhichonly 3species parasitize fishes of the family Auchenipteridae (Siluri-formes):  Cucullanus brevispiculus  Moravec, Kohn and Fernandes,1993;  C. heliomartinsi   Moreira, Rocha and Costa, 2000; and  C. pinnai  Travassos,ArtigasandPereira,1928(Lo´pez-Caballeroetal.,2009). Species of   Cucullanus  share many morphological features,such as the distribution the male genital papillae; however, thesrcinal descriptions of some species are unclear and comparisonsamong species are difficult. Consequently, among some authorswhen describing a new species, it has became common practice toonly compare these new species to those from the samezoogeographical regions (Moravec et al., 2005; Caban˜as-Carranzaand Caspeta-Mandujano, 2007) or the same host group (Gonza´les-Solı´s et al., 2007; Moravec et al., 2008). MATERIALS AND METHODS In total, 189  Ageneiosus ucayalensis  Castelneau, 1855 caught in theestuary of the Guama´ River and Guajara´ Bay, Bele´m, Para´, Brazil, wereacquired from a fisherman, between November 2006 and August 2008.Fishes were transported alive in plastic containers to the laboratory,necropsied, and worms were collected alive. Worms were subsequentlyrinsed in phosphate-buffered saline (PBS), fixed in alcohol-formaldehyde-acetic acid (AFA) at 60 C, and stored in the same solution. For lightmicroscopy, worms were cleared in lactophenol and examined with aBX41 microscope (Olympus, Tokyo, Japan). Figures were drawn with theaid of a camera lucida attached to the microscope. For scanning electronmicroscopy (SEM), worms were rinsed in PBS, fixed in AFA at 60 C, post-fixed for 2 hr in 1 %  osmium tetroxide, dehydrated in an ethanol series,critical point dried using CO 2 , mounted on a metallic stub, and goldsputter coated at a thickness of 5 A˚ . The specimens were examined andphotographed using a LEO 1459 VP scanning electron microscope. Allmeasurements are in micrometers unless otherwise noted. DESCRIPTION Cucullanus ageneiosus   n. sp. (Figs. 1–22) Diagnosis:  Nematode medium size, opaque white when alive. Malessmaller than females, anterior region similar in morphology in both sexes(Figs. 1–4, 9, 10). Cuticle with fine transversal striations (Figs. 11, 18).Cephalic extremity rounded, oral aperture slit-like, dorsoventrallyelongated, surrounded by cuticular ring (collarette), internally delimitedby row of small teeth-like structures (Figs. 9, 10). Cephalic papillae sub-median, 4 in number. Pair of lateral amphids present (Fig. 10). Cervicallateral alae absent (Figs. 1–4). Muscular esophagus divided in 2 distinctand well developed regions, anterior region well sclerotized forming buccalpseudocapsule, also called esophastome, and posterior region presentingstrong musculature terminating just posterior to nervous ring (Figs. 3, 4).Glandular esophagus claviform, opening into intestine through strongvalve (Figs. 3, 4). Excretory pore and deirids at mid-region of glandularesophagus (Figs. 3, 4, 11, 12). Conical tail with pointed tip (Figs. 1, 2, 5, 7,14, 16, 18, 20). Male  ( based on 4 specimens; measurements of holotype in parentheses ) : Body 5.9 ± 1.94 (3.65–8.94) (8.94) long and 0.23 ± 0.02 (0.21–0.26) (0.26)wide (Fig. 2). Esophastome 0.07 ± 0.002 (0.07–0.08) (0.07) long and 0.1 ± 0.01 (0.09–0.12) (0.09) wide. Esophagus 0.54 ± 0.02 (0.51–0.57) (0.57) longand 0.1  ±  0.009 (0.09–0.1) (0.11) wide, corresponding to 10.2 %  of totalbody length. Nerve ring 0.30  ±  0.007 (0.30–0.31) (0.30) from anteriorextremity. Deirids and excretory pore 0.58  ±  0.02 (0.53–0.62) (0.56) and0.59  ±  0.02 (0.54–0.61) (061), respectively, from anterior extremity.Ventral precloacal sucker well developed and situated at 0.06  ±  0.09(0.55–0.76) (0.76) from posterior extremity (Fig. 7). Cloacae withoutelevated lips (Figs. 7, 16, 17). Gubernaculum 0.09  ±  0.009 (0.08–0.10)(0.08) long; spicules very small, slightly sclerotized, subequal 0.16 ± 0.01(0.14–0.18) (0.18) long and 0.18 ± 0.02 (0.16–0.22) (0.19) wide (Figs. 7, 8).Caudal papillae consisting of 1 medial adcloacal papilla and 10 pairs of papillae, 5 pairs precloacal, all subventral (pair 1 anterior to ventralsucker; pairs 2, 3, 4, and 5 after the sucker, the first 3 pairs distributedequidistant, pairs 4 and 5 immediately preceding cloacal opening) and 5pairs postcloacal (pairs 6, 7, and 10 subventral, pairs 8 and 9 laterals)(Figs. 7, 16, 17, 20, 21). Pair of lateral phasmids, situated immediatelyposterior to 10th pair of papilla, 0.07  ±  0.01 (0.06–0.09) (0.07) fromposterior extremity (Figs. 7, 20–22). Tail length 0.25  ±  0.01 (0.23–0.26)(0.26) (Figs. 7, 16). Female  ( based on 3 specimens; measurements of allotype in parentheses ) : Body 10.8 ± 0.8 (10–12) (12) long and 0.26 ± 0.03 (0.23–0.31) (0.31) wide(Fig. 1). Esophastome 0.08 ± 0.04 (0.08–0.09) (0.09) long and 0.1 ± 0.003(0.1–0.019 (0.103) wide. Esophagus 0.97  ±  0.02 (0.94–1) (0.98) long and0.12 ± 0.01 (0.1–0.14) (0.14) wide, corresponding to 17.5 % of total bodylength (Figs. 1, 4). Nerve ring and deirids 0.33 ± 0.009 (0.32–0 34) (0.32)and 0.66 ± 0.07 (0.57–0.75) (0.65) from anterior end, respectively (Figs. 4,12). Excretory pore 0.71  ±  0.07 (0.65–0.80) (0.69) from anterior end(Figs. 4, 11). Vulva with slightly projected lips (Figs. 1, 6, 13) post-equatorial 3.73  ±  0.07 (0.63–0.80) (4.7) from posterior end. Vaginamuscular, directed anteriorly. Uteri amphidelphic (Figs. 1, 6). Anus 0.28 ±  0.02 (0.25–0.31) (0.27) from posterior end (Figs. 5, 14, 15). Phasmids0.07  ±  0.006 (0.06–0.07) (0.07) from posterior extremity. Conical tail(Figs. 5, 14, 18, 19). Taxonomic summary Type host: Ageneiosus ucayalensis  Castelnau, 1855 (Auchenipteridae,Siluriforme). Common name:  Mandube´. Site of infection:  Small intestine. Type locality:  Guajara´ Bay (estuary), Bele´m, Para´, Brazil (1 u 15 9  –1 u 29 9 S,48 u 32 9  –48 u 29 9 W). Type specimens:  Colec¸a˜o Helmintolo´gica do Instituto Oswaldo Cruz,FIOCRUZ, Rio de Janeiro (holotype: CHIOC 35657-a and allotype:CHIOC 35657-b).Received 28 February 2009; revised 15 April 2009, revised 22 September;accepted 13 November 2009.*To whom correspondence should be addressed. DOI: 10.1645/GE-2081.1 J. Parasitol.,  96(2), 2010, pp. 389–394 F American Society of Parasitologists 2010 389  F IGURES  1–8. Light microscopy of   Cucullanus ageneiosus  n. sp. ( 1 ) General view of female. Bar  5  400  m m. ( 2 ) General view of male. Bar  5  400  m m.( 3 ) Male anterior region, showing esophastome, muscular and glandular esophagus, nervous ring, excretory pore, and deirids. Bar  5  100  m m. ( 4 ) Femaleanterior region, showing esophastome, muscular and glandular esophagus, nervous ring, excretory pore, and deirids. Bar  5  100  m m. ( 5 ) Lateral view of the female posterior region, showing anus. Bar  5  100  m m. ( 6 ) Detail of the vulvar region, showing vulva, muscular vagina, and distal region of theamphidelphic uterus with eggs. Bar  5  100  m m. ( 7 ) Lateral view of the male posterior region, cloaca, spicules, gubernaculum, cloacal sucker, and caudalpapillae. Bar  5  100  m m. ( 8 ) Detail of the lateral view of the spicules and gubernaculum. Bar  5  50  m m. 390 THE JOURNAL OF PARASITOLOGY, VOL. 96, NO. 2, APRIL 2010  F IGURES  9–17. Scanning electron microscopy of   Cucullanus ageneiosus  n. sp. ( 9)  Ventral view of the anterior end, showing oral aperture (oa)surrounded by a cuticular ring (cr), distal end of the esophastome and cephalic papillae (cp). Bar  5  25  m m. ( 10 ) Apical view of the anterior end showingoral aperture, 2 pairs of cephalic papillae (cp), amphids (am), and cuticular ring (cr) with an internal row of denticles (arrow). Bar  5  15  m m. ( 11 ) Detail of the excretory pore (ep) surrounded by transverse cuticular striations. Bar  5  3  m m. ( 12 ) Detail of the deirid. Bar  5  5  m m. ( 13 ) Detail of the vulvar aperture(v). Bar  5  25  m m. ( 14 ) Lateral view of female tail. Note the conical shape with thin tip end. Phasmid (ph), anus (an). Bar  5  50  m m. ( 15 ) Detail of anusaperture. Bar  5  15  m m. ( 16 ) Lateroventral view of the male posterior end, cloacal aperture (cl), caudal papillae: 1 pair pre sucker (psp), 4 pairs pre-cloacal (pcp), 3 pairs post-cloacal ventral (pcvp), 2 pair of post-cloacal lateral (pclp). Bar  5  60  m m. ( 17 ) Detail of the pre (pcp) and post-cloacal papillae(pcvp) and the unpaired pre-cloacal papillae and cloacal aperture (cl). Bar  5  12  m m. GIESE ET AL.— CUCULLANUS AGENEIOSUS   N. SP. FROM BRAZIL 391  F IGURES  18–22. Scanning electron microscopy of   Cucullanus ageneiosus  n. sp. male and female tails. ( 18 ) Lateral view of female posterior end,showing conical tail and phasmid (ph). Bar  5  15  m m ( 19 ) Detail of rounded structure of female phasmid. Bar  5  2  m m. ( 20 ) Lateral view of final third of male tail. Note the 7p, 8p, 9p, and 10p papillae and phasmid (ph). Bar  5  15  m m. ( 21 ) Frontal view of 9p papilla round shaped and central button-like tip,visible in lateral view of 10p papilla. Note the phasmid (ph) is situated immediately posterior to 10p papilla. Bar  5  15  m m. ( 22 ) High magnification of male phasmid rounded shaped. Bar  5  2  m m. 392 THE JOURNAL OF PARASITOLOGY, VOL. 96, NO. 2, APRIL 2010
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