Vol. 3 (2): 127-141, 2019

Occurrence of Six Larval Species from Marine Fishes of Iraq

Hayder A. H. Al-Hasson1, Khalidah S. Al-Niaeem2* & Suzan A. Al-Azizz3

1Education Directorate of Basrah, Basrah, Iraq 2Department of Fisheries and Marine Resources, College of Agriculture, University of Basrah, Basrah, Iraq 3Department of Microbiology and Veterinary Parasitology, College of Veterinary Medicine, University of Basrah, Basrah, Iraq

*Corresponding author: [email protected]

Abstract: Six larval nematode species belonging to two different classes: Adenophorea (family Trichosomoididae) and class Chromadorea (families Raphidascaridae and Cucullanidae) were collected from five fish species from northwest of the Arab Gulf during the period from October 2014 till July 2015. sp. was found attached on the mesenteries of two bar sea bream Acanthopagrus bifasciatus. Hysterothylacium sp. BA larva was found in intestine of Argyrops spinifer and Acanthopagrus arabicus, Hysterothylacium sp. BC larvae were recorded from intestine of A. spinifer, Ephippus orbis and Sphyraena obtusata. Cucullanus sp. 1 was recorded from intestine of A. spinifer, while Cucullanus sp. 2 and Proleptinae gen. sp. type BA larvae were recorded from intestine of A. arabicus. The record of Huffmanela sp. represents the first record in Iraq and A. bifasciatus is now considered as a new host for this parasite in the Arab Gulf.

Keywords: Nematode larvae, Perciform fishes, Iraqi marine waters.

Introduction The pathogenesis of parasitic diseases depends upon several factors such as species and numbers of parasites, size of the host species and its health status, and site of infection (Barson, 2004; Omeji et al., 2011). is extremely common in nature and is one factor that can contribute significantly to ecosystem stability and diversity, although most parasites are small in body size they have been found to account for a substantial amount of biomass within an ecosystem, which can lead to strong effects on community structure, composition, and energy flow (Omeji et al., 2011; Cook, 2012). Adults of Huffmanela extra-intestinal deposit eggs in the skin, gill, somatic musculature, swim bladder, bone and mesentery (Ruiz & Bullard, 2013). Six of Huffmanela plus one unnamed species (Worsham et al., 2016) reportedly infect the skin only (Bullard et al., 2012, Ruiz & Bullard, 2013) and most Huffmanela tissue-dwelling nematodes lay numerous eggs that generally form black spots in the flesh or other tissues of fishes. After being laid, eggs finish their maturation, become dark, and are often noticed only when the adults are dead and unavailable for study (Moravec, 2001; Justine, 2007). Kardousha (1992) record two species of larval nematodes including Hysterothylacium sp. MB that found in four fishes in United Arab Emirate (UAE) coasts. El-Naffar et al. (1992) recorded three genera of nematodes including Hysterothylacium from some fishes in UAE coasts. González-Solís et al. (1997) isolated Hysterothylacium sp. larval stage from Scomberomorus guttatus from Iranian marine waters.

Received Apr. 18, 2019, accepted July 25, 2019

128 Al-Hasson et al.

Al-Salim & Ali (2010) recorded eight species of nematodes, Hysterothylacium Ward et Magath, 1917 from five species of marine bony fishes and three marine from northwest of Arab Gulf at Khor Al-Ummia. Ali & Al-Salim (2013) recorded five larval nematode species including Proleptinae gen. sp. BA larva from largescale tonguesole Cynoglossus arel from Iraqi marine waters. According to Mhaisen et al. (2018), out of 253 parasite species infecting marine fishes of Iraq, the nematodes contribute to 18. 6% of such parasitic fauna.

Materials and Methods A total of 55 fish were collected by fishermen using trawl net monthly, during the period from October 2013 to the end of July 2014, which belong five species of perciform fishes: Acanthopagrus arabicus (Iwatsuki, 2013), Acanthopagrus bifasciatus (Forsskål, 1775), Argyrops spinifer (Forsskål, 1775), Ephippus orbis (Bloch, 1787) and Sphyraena obtusata (Cuvier, 1829). They were taken from Iraqi marine waters, Northwest Arab Gulf (latitudes 47° 30′ to 48° 15′ N; 30° 50′ to 30° 00′ E). Fishes were identified according to Carpenter et al. (1997) and their scientific names were updated according to Froese & Pauly (2019). Vigorously moving worms, separated from mesenteries and intestine of their hosts with a pipette and samples for light microscopy, were handled according to Moravec (1994).

Results The following is a systematic scheme of the detected nematodes (according to GBIF, 2019) from the investigated fishes of the present study in Iraqi marine waters.

Phylum Nematoda Class Adenophorea Order Enoplida Family Trichosomoididae Huffmanela sp. Class Secernentea Order Ascaridida Family Ascarididae Hysterothylacium sp. type BA Hysterothylacium sp. type BC Family Cucullanidae Cucullanus sp. 1 Cucullanus sp. 2 Order Spirurida Family Physalopteridae Proleptinae gen. sp. BA

The following is a brief account on the occurrence of these nematodes which were recorded from marine fishes of Iraq in the present investigation.

Huffmanela sp. )Plates 1 & 2 and Figures 1 & 2( Host: Acanthopagrus bifasciatus. Site of infection: Mesenteries. Prevalence of infection: 2%. Mean intensity: 2.

Occurrence of six larval nematode species from marine fishes of Iraq 129

Description and measurements (in micrometers): Female: Single female specimen, probably immature. Body length 10201, maximum width 80, width of cephalic end 34. Length of entire esophagus 4830 (47% of body length). Muscular esophagus with anterior cuticularised part, 443 in length. Region of vulva damaged, not illustrated. Immature eggs in uterus. Vulva 1834 prequatorial 17.9% from anterior end. Tail with rounded end (Plate 1 & Figure 1). Male: Single male, 6801 in total length, 24 in maximum width, width of cephalic end 10, of posterior end 16. Length of entire esophagus 2584 (38% of body length), length of muscular esophagus 354, length of posterior part of esophagus 4420. Nerve ring 1294 from anterior extremity. Length of stichocytes 78-127 in middle region of stichosome, terminal stichocyte 155 in length (Plate 2 & Figure 2).

Plate 1: Huffmanela sp. Female: A- Anterior body, B- Posterior end. 100X.

130 Al-Hasson et al.

Figure 1: Huffmanela sp. Female: A- Anterior body, B- Posterior end. Scale bar= 1.4 mm. Occurrence of six larval nematode species from marine fishes of Iraq 131

Plate 2: Huffmanela sp. Male: A- Anterior body, 400X, B- Posterior end. 100X.

132 Al-Hasson et al.

Figure 2: Huffmanela sp. Male: A- Anterior body, B- Posterior end. Scale bar= 1.4 mm.

1.4

Occurrence of six larval nematode species from marine fishes of Iraq 133

Hysterothylacium sp. type BA )Plate 3( Hosts: Argyrops spinifer and Acanthopagrus arabicus. Site of infection: Intestine. Prevalence of infection: 11% and 50% for A. spinifer and A. arabicus, respectively. Mean intensity: 3 and 2 for A. spinifer and A. arabicus, respectively.

Plate 3: Hysterothylacium sp. BA: A- Entire body, B- Posterior end. 100X.

134 Al-Hasson et al.

Hysterothylacium sp. BC (Plate 4( Hosts: Argyrops spinifer, Ephippus orbis and Sphyraena obtusata. Site of infection: Intestine. Prevalence of infection: 16%, 10% and 9% for A. spinifer, E. orbis and S. obtusata, respectively. Mean intensity: 1, 1 and 19 for A. spinifer, E. orbis and S. obtusata, respectively.

Plate 4: Hysterothylacium sp. BC: Anterior body. 40X.

Cucullanus sp. 1 (Plate 5 and Figure 3( Host: Argyrops spinifer. Site of infection: Intestine. Prevalence of infection: 16%. Mean intensity: 1.

Description and measurements (in micrometers, based on four females): Length of body 10908, maximum width 613 close to vulva. Length of esophagus 1546, representing 7.1-10.5 (14.17% of whole body length). Length of pseudobuccal capsule 338 and width 700. Width of esophagus at anterior end 287 and at posterior end 252. Nerve ring and excretory pore 173-510 (342) and 597-735 (666) from anterior end, respectively. Length of vulva 70 and width 105. Vulva lips protruded and elevated, postequatorial 12572 from anterior extremity, representing 95.86% of body length. Vagina muscular, directed slightly toward anterior end then back to posterior end. Mature eggs oval.

Occurrence of six larval nematode species from marine fishes of Iraq 135

Plate 5: Cucullanus sp. 1: A- Anterior body, B- Posterior end. 400X.

136 Al-Hasson et al.

Figure 3: Cucullanus sp. 1: A- Anterior body, B- Posterior end, C and D- Egg. Scale bar= 0.4 mm for A, B and C and 0.1 mm for D.

Occurrence of six larval nematode species from marine fishes of Iraq 137

Cucullanus sp. 2 (Plate 6( Host: Acanthopagrus arabicus. Site of infection: Intestine. Prevalence of infection: 50%. Mean intensity: 3.

Plate 6: Cucullanus sp. 2: A- Anterior body, B- Posterior end. 400X.

Proleptinae gen. sp. type BA larva (Plate 7( Host: Acanthopagrus arabicus. Site of infection: Intestine. Prevalence of infection: 25%. Mean intensity: 8.

138 Al-Hasson et al.

Plate 7: Proleptinae gen sp. whole body. 40X.

Discussion Huffmanela sp. By possession of stichosome, single row of stichocytes, absence of spicule and pseudobursa in the male, vulva opens near posterior end of esophagus in female, the specimens fall in the genus Huffmanela Moravec, 1987. One male and one female with empty uterus (immature or laid mature female) were detected in the present study. However, both specimens were not in a good condition for full description. It is worth to mention that all 20 valid species of Huffmanela that described from fishes in the world were never reported from fishes of the family Sparidae. Justine & Iwaki (2014) listed eight families of the order Perciformes (, Labridae, , , Mullidae, Nemipteridae, and Xiphiidae), three families of the order Tetradontiformes (Balistidae, Monacanthidae and ), one family of the order Pleuronectiformes (Cynoglossidae), one family of sharks (Carcharhinidae), one family of the order Atheriniformes (Atherinopsidae), one family of the order Beloniformes (Exocoetidae), one family of the order Gadiformes (Gadidae), one family of the order Ophidiiformes (Ophidiidae), one family of the order Scorpaeniformes (( and one family of the order Anguiliformes (). Ali et al. (2014) referred that the superfamily Trichinelloidea, to which Huffmanela sp. of the family Trichosomoididae belongs, is represented in fishes of Basrah province with two species of the genera and Pseudocapillaria and both were recorded from freshwater fishes. Hence, Huffmanela sp. of the present investigation is considered here as the first record in Iraq and A. bifasciatus is considered as a new host record for Huffanela sp. in the world.

Occurrence of six larval nematode species from marine fishes of Iraq 139

Hysterothylacium sp. BA Al-Salim & Ali (2010) designed a key for identification of eight different larval types and one adult of Hysterothylacium which parasitize marine fishes in Iraq. Hysterothylacium sp. BA is recognized by possessing the ratio of caecum to appendage as 1: 8.6, ratio of esophagus to appendage as 1:5 and presence of fine lateral alae. Ali (2008) recorded Hysterothylacium sp. BA from four different families (Hemiscyliidae, Sphyrnidae, Sparidae and Cynoglossidae) from Khor Al-Ummia off Iraq. Two new hosts from Sparidae were added in this study (A. arabicus and A. spinifer) for Hysterothylacium sp. type BA in Iraq.

Hysterothylacium sp. BC Hysterothylacium sp. type BC larva is recognized by having a long appendage, about 34% of body length, ratio of caecum to appendage as 1:21.6, caecum to esophagus as 1:7.2 as well as appendage as a about three times as that of esophagus. Ali (2008) recorded this larval type from two species of marine fishes (Cynoglossus arel and Chiloscylium arabicum). Hence, three fish species (A. spinifer, E. orbis and S. obtusata) are now considered as new hosts for Hysterothylacium sp. type BC in Iraq.

Cucullanus sp. 1 Members of the genus Cucullanus generally parasitize freshwater, estuarine and marine fishes, and rarely aquatic turtles. This genus is mostly composed of species whose members are of medium to large size with thick cuticle. They mostly have an identical morphology and some of them are poorly described, making their comparison difficult. However, some authors have defined criteria for their identification (Moravec & Ali, 2005; González-Solis et al., 2007; Park & Moravec, 2008). Despite the availability of four females of Cucullanus sp. 1 and because no male worms of this specimen were found, this nematode could not be identified to species level. This unidentified species is recorded here for the first time in Iraq and A. spinifer is considered as a new host for this species in Iraq.

Cucullanus sp. 2 This nematode could not be identified to species because no male worms were found. Cucullanus sp. 2, isolated from A. arabicus of present study, agreed in morphological characters with Cucullanus sp. of Ali (2008) and Al-Salim & Ali (2011) from Lethrinus nebulosus off Iraqi marine waters. The present unidentified species differ from Cucullanus sp.1 in the ratio of esophagus to body length and the distance of the nerve ring from the anterior end of the body. A. arabicus is considered now as a new host for this parasite in Iraq.

Proleptinae gen. sp. BA larvae Morphological characters of the present specimens agree with those of the subfamily Propletinae, and the generic identification of these larvae is impossible to be establish because the distinguished characters among different genera, such as situation of the vulva and type of spicules are found in adults only (Chabaud, 1975; Anderson et al., 2009). The present specimens belong to one of the three genera Paraleptus, Proleptus and Heliconema, while Bulbocephalus is excluded from probability of identification due to the lack of the cephalic collar. The final host of Paraleptus and Proleptus are Chondrichthys, while Anguilliformes in most time are the final host of Heliconema (Moravec et al., 2007). Ali (2008) described the adult of Paraleptus sp. from Arabian carpet C. arabicum from Iraqi marine waters. The recent unidentified species was later redescribed and studied with the aid of SEM by González-Solís & Ali (2015) as Paraleptus chiloscyllii Yin & Zhang, 1983. Ali (2008) and Ali & Al-Salim (2013) recorded this type of larva from Cynoglossus arel from Khor Al- 140 Al-Hasson et al.

Ummiah, north-west of the Arab Gulf. The measurements of the present larva agreed with those of the same parasite of the present investigation, and hence A. arabicus is considered as a new host record in Iraq.

References Ali, A.H. (2008). of helminth parasites in some marine and freshwater fishes and the relation of some of it's with their final hosts in southern of Iraq. Ph. D. Thesis, Coll. Agric., Univ. Basrah: 336 pp. (In Arabic). Ali, A.H. & Al-Salim, N.K. (2013). First record of five larval nematode species from fishes of Iraq. Basrah J. Agric. Sci., 26 (Spec. Issue 1): 142-156. Ali, A.H.; Mhaisen, F.T. & Khamees, N.R. (2014). Checklists of nematodes of freshwater and marine fishes of Basrah province, Iraq. Mesopot. J. Mar. Sci., 29 (2): 71-96. Al-Salim, N.K. & Ali, A.H. (2010). Description of eight nematode species of the genus Hysterothylacium Ward et Magath, 1917 parasitized in some Iraqi marine fishes. Basrah J. Agric. Sci., 23 (Spec. Issue 2): 115-137. Al-Salim, N.K. & Ali, A.H. (2011). First record of three nematode species parasitized some marine fishes in Iraq. Basrah J. Res. (Sci.), 37 (4E): 17-26. Anderson, R.C.; Chabaud, A.G. & Willmott, S. (2009). Keys to the nematode parasites of vertebrates: Archival volume. CAB Int., Wallingford: 463 pp. Barson, M. (2004). Endoparasites of the sharptooth catfish, Clarias garirpinus (Burchell), from the Rietvlei dam, Sesmyl Spurit system, South Africa. M. Sc. Thesis, Coll. Sci., Univ. Rand Afrikaans: 53 pp. Bullard, S.A.; Ruiz, C.F.; McElwain, A.; Murray, M.J.; Borucinska, J.D. & Benz, G.W. (2012). Huffmanela cf. carcharhini (Nematoda: Trichosomoididae: Huffmanelinae) from skin of a , Carcharhinus plumbeus, in the Pacific Ocean. J. Parasitol., 98 (2): 333-340. Carpenter, K.E.; Krupp, F.; Jones, D.A. & Zajonz, U. (1997). FAO species identification field guide for fishery purposes living marine resources of Kuwait, Eastern Saudi Arabia, Bahrain, Qatar, and the United Arab Emirates. FAO, Rome: viii + 293 pp. + XVII pls. Chabaud, A.G. (1975). Keys to genera of the order Spirurida. Part I. Camallanoidea, Dracunculoidea, Gnathostomatoidea, Physalopteroidea, Rictularoidea and Thelazioidea. In: Anderson, R.C.; Chabaud, A.G. & Willmott, S. (eds.), CIH Keys to the nematode parasites of vertebrates, CAB vol. 3. Farnham Royal, Bucks (UK): 27 pp. Cook, C.W. (2012). The early life history and reproductive biology of Cymothoa excisa, a marine isopod parasitizing Atlantic croaker, (Micropogonias undulatus), along the Texas coast. M. Sc. Thesis, Fac. Grad. Sch., Univ. Texas at Austin: 71 pp. El-Naffar, M.K.; Gobashy, A.F.; El-Etraby, S. & Kardousha, M.M. (1992). General survey of helminth parasite genera of Arabian Gulf fish (coasts of United Arab Emirates). Arab Gulf J. Sci. Res., 10 (2): 99-110. Froese, R. & Pauly, D. (eds.) (2019). FishBase. World Wide Web electronic publication. www.fishBase.org. (Version 02/2019). GBIF (2019). Global Biodiversity Information Facility, on-line database, http://www.gbif.org. (Accessed 26 June 2019). González-Solís, D. & Ali, A.H. (2015). Redescription of Paraleptus chiloscyllii Yin et Zhang, 1983 (Nematoda: Physalopteridae) from the Arabian carpetshark Chiloscyllium arabicum (Chondrichthyes: Hemiscylliidae) off Iraq. Acta Parasitol., 60 (4): 759-766. González-Solís, D.; Moravec, F. & Coad, B.W. (1997). Some nematode parasites of fishes from southwestern Iran. Zool. Mid. East, 15 (1): 113-119.

Occurrence of six larval nematode species from marine fishes of Iraq 141

González-Solis, D.; Tuz-Paredes, V.M. & Quintal-Loria, M.A. (2007). Cucullanus pargi sp. n. (Nematoda: Cucullanidae) from the grey snapper Lutjanus griseus off the southern coast of Quintana Roo, Mexico. Fol. Parasitol., 54 (3): 220-224. Justine, J.-L. (2007). Huffmanela spp. (Nematoda, Trichosomoididae) parasites in coral reef fishes off New Caledonia, with descriptions of H. ballista n. sp. and H. longa n. sp. Zootaxa, 1628: 23-41. Justine, J.-L. & Iwaki, T. (2014). Huffmanela hamo sp. n. (Nematoda: Trichosomoididae: Huffmanelinae) from the dagger-tooth pike conger cinereus off Japan. Fol. Parasitol., 61 (3): 267-271. Kardousha, M.M. (1992). Helminth parasite larvae collected from Arabian Gulf fish (coasts of the United Arab Emirates). I: Anisakid larvae (Nematoda: Anisakidae). Jpn. J. Parasitol., 41 (6): 464-472. Mhaisen, F.T.; Ali, A.H. & Khamees, N.R. (2018). Marine fish parasitology of Iraq: A review and checklists. Biol. Appl. Environ. Res., 2 (2): 231-297. Moravec, F. (1994). Parasitic nematodes of freshwater fishes of Europe. Academia and Dordrecht Kluwer Acad. Publ., Prague: 473 pp. Moravec, F. (2001). Trichinelloid nematodes parasitic in cold-blooded vertebrates. Academia, Praha: 432 pp. Moravec, F. & Ali, A.H. (2005). Two new species of Philometra (Nematoda: Philometridae) from needlefishes (Belonidae) in Iraq, with a key to Philometra spp. parasitic in the host’s subcutaneous tissue, fins and musculature. Fol. Parasitol., 52: 267-273. Moravec, F.; Taraschewski, H.; Anantaphruti M.T.; Maipanich, W. & Laoprasert, T. (2007). Heliconema longissimum (Ortlepp, 1923) (Nematoda: Physalopteridae) from Pisodonophis boro (Teleostei: Ophichthidae) in Thailand, with remarks on the taxonomy of the Proleptinae Schulz, 1927. Syst. Parasitol., 66 (1): 73-80. Omeji, S.; Solomon, S.G. & Idoga, E.S. (2011). A comparative study of the common protozoan parasites of Clarias gariepinus from the wild and cultured environments in Benue State, Nigeria, J. Parasitol. Res., 2011. Article ID 916489, 8 pages. Park, J.-K. & Moravec, F. (2008). Redescription of Cucullanus robustus (Nematoda: Cucullanidae) from the conger Conger myriaster of Korea. Zootaxa, 1729: 1-7. DOI: 10.5281/zenodo.181264. Ruiz, C.F. & Bullard, S.A. (2013). Huffmanela markgracei sp. n. (Nematoda: Trichosomoididae) from buccal cavity of , Rhizoprionodon terraenovae (Carcharhiniformes: Carcharhinidae), in the northwestern Gulf of Mexico off Texas. Fol. Parasitol., 60 (4): 353-358. Worsham, M.L.D.; Huffman, D.G.; Moravec, F. & Gibson, J.R. (2016). The life cycle of Huffmanela huffmani Moravec, 1987 (Nematoda: Trichosomoididae), an endemic marine-relict parasite of Centrarchidae from a Central Texas spring. Fol. Parasitol., 63: 020. DOI: 10.14411/fp.2016.020.