Travaux du Muséum National d’Histoire Naturelle © 30 Juin Vol. LIV (1) pp. 63–68 «Grigore Antipa» 2011 DOI: 10.2478/v10191-011-0004-4

INFESTATION OF THE PUMPKINSEED LEPOMIS GIBBOSUS (TELEOSTEI: CYPRINIDAE) BY THE CYPRINACEA (CRUSTACEA) – SOME ECOLOGICAL ASPECTS

MALA-MARIA STAVRESCU-BEDIVAN, OANA P. POPA, FLORIN T. AIOANEI, LUIS O. POPA

Abstract. The copepod Linnaeus, 1758 is known by the large spectrum of fish hosts which it infests, and to which it can cause haemorrhages and ulcerations. The paper focuses on some aspects of the relation between the Pumpkinseed sunfish and this copepod: infestation parameters, preferred attaching situs, relationship between the host size and parasitisation degree, as well as the visible effects of this parasite left on the fish body, in the Moara Domneascã Lake, Romania. Résumé. Le copépode Lernaea cyprinacea Linnaeus, 1758 est connu pour son grand spectre de poissons qu’il infeste et auxquels il peut produire des hémorragies ainsi que des ulcérations. Ce travail se concentre sur quelques aspects de la relation entre la perche arc-en-ciel et ce copépode: les paramètres de l’infestation, les sites d’attachement préférés, la relation entre les dimensions du poisson et le degré d’infestation par le parasite, ainsi que les effets visibles provoqués par ce parasite sur la surface du corps du poisson provenant du Lac Moara Domneascã (Roumanie). Key words: infestation, attachment site, Lepomis gibbosus, Lernaea cyprinacea, Moara Domneascã Lake.

INTRODUCTION The present study was performed in the frame of a project aimed at the evaluation of the diversity of some ectoparasitic metazoans on the prevalent fish species of the lentic ecosystem from Moara Domneascã. The study site is located in the North-East of Bucharest (44°50’N, 26°13’E, at an altitude of 90 m), on the right side of national highway Bucharest – Constanþa (Fig. 1). The only water source of the area is Moara Domneascã Lake, belonging to Pasãrea lake chain which opens in the Argeº River. The Moara Domneascã Lake is used for irrigation of the neighbouring agricultural areas and also for pisciculture. Lernaea cyprinacea Linnaeus, 1758 (Copepoda: ), called also “the anchor worm”, is a parasite of freshwater fishes and amphibians tadpoles all over the world. The species is naturally distributed in Europe, Africa, Central Asia, southern regions of West Siberia and is introduced into North and South America and into eastern and western Australia by the importation of tropical fish species. Only the adult female of this lernaeid species is parasitic. It can reach 9 mm long (Tirmizi, 2003). Immature forms of females and males are free swimming. After mating, the male dies and the female looks for a host fish. The parasite fixes on the fish body by an “anchoring” device which develops under tegument, especially in the muscular tissue. Then, the parasite leaves the fish, dragging bags full of eggs behind. Infestations can lead to injuries which can infect themselves with bacteria or fungi. For the small sized fish specimens, infections can be lethal. Ulcerations hardly heal. 64 MALA-MARIA STAVRESCU-BEDIVAN, OANA P. POPA, FLORIN T. AIOANEI, LUIS O. POPA

Fig. 1 - Moara Domneascã (see the black arrow) location on Google Earth map (14.2 km altitude from the observer visual level).

The literature reports about L. cyprinacea infecting freshwater fish species in Romania are scarce, usually representing veterinary case reports: Boz & Vulpe (1998), Vulpe et al. (2001), Hangan et al. (2008), Patriche et al. (2009). In this study, in Moara Domneascã Lake, the species was found to infest several host fish species: Carassius auratus, Rutilus rutilus, Pseudorasbora parva and Lepomis gibbosus. We analysed aspects of the infestation with L. cyprinacea in the last mentioned fish species because we had a rather large sample of Pumpkinseed sunfish. L. gibbosus is a fish species with a life cycle of almost eight years. The genus Lernaea, as a possible parasite for L. gibbosus from Romanian fauna, is mentioned since 1964 by Bãnãrescu (Gavriloaie & Chiº, 2007). To our knowledge this is the first ecological approach regarding the distribution of this parasite species on the surface of the host species, in the Romanian fauna.

MATERIAL AND METHODS The studied fish community was sampled in June 2008 from two sites of the Moara Domneascã Lake, by electrofishing with a Samus 720 MP device (Iorgu et al., 2009). One sample was taken near the USAMV Research Station, Bucharest, and the second one near Moara Domneascã locality, from an area which seemed to be damaged by organic pollution. Both sample stations had similar species composition, with the same prevalent species (Scardinius erythrophthalmus, Rutilus rutilus, Carassius auratus, Lepomis gibbosus etc.). We analysed the parasitic fauna of 80 specimens of Pumpkinseed sunfish (Lepomis gibbosus), collected as mentioned above. Immediately after sampling, the fish specimens were put in 5% formaldehyde and transported to the laboratory for INFESTATION OF LEPOMIS GIBBOSUS BY LERNAEA CYPRINACEA 65 analysis. Here, the parasite specimens were removed and preserved in 1.5 ml centrifuge tubes containing 70% alcohol. Each fish specimen was studied both at the level of the branchial cavities and at the external surface of their body, head, fins and skin. Information regarding the parasite distribution at the host level was noted in the parasitic topography card. For each fish specimen the total length and the standardised length were measured with a calliper to the nearest 0.1mm. The correlation between the host fish specimens and the corresponding parasite number was studied with the Bravais-Pearson correlation coefficient. The morphological identification of the parasite species was performed according to Bauer (1987). The following infestation parameters were calculated according to Bush et al. (1997): prevalence, mean intensity and mean abundance.

RESULTS AND DISCUSSIONS The parasitological study showed that the Pumpkinseed sunfish (Lepomis gibbosus) was infested by the copepod Lernaea cyprinacea (Fig. 2).

Fig. 2 - Lepomis gibbosus: a specimen infested by Lernaea cyprinacea.

The infestation parameters of the host specimens with adult females of L. cyprinacea were prevalence (58.75%), mean intensity 1.66 (min. 1 – max. 6) and mean abundance (0.975). The distribution of the parasite on the surface of the host body showed that the 78 parasitic specimens were fixed on the host surface, as follows: head (43.59%), fins (33.33%) and skin (23.08%) (Fig. 3). The bases of the anal, pectoral and dorsal fins were common attachment sites. According to Pérez-Bote (2010), the fins offer the greatest protection from water currents and the tissues near the bases may be easily penetrated by the parasite. We performed a finer scale analysis at the level of the host head, as the most parasitized area, and we observed the following spatial distribution of the parasites at this level: opercular apparatus (58.82%), around the mouth (26.47%), gills (11.76%), around the eyes (2.94%) (Fig. 4). Several L. gibbosus specimens without L. cyprinacea presented ulcerations on the opercular apparatus and scars at the base of their dorsal fins, caused probably by previous infestations. Similar findings about ulcerations at the site of L. 66 MALA-MARIA STAVRESCU-BEDIVAN, OANA P. POPA, FLORIN T. AIOANEI, LUIS O. POPA

Fig. 3 - Lernaea cyprinacea: adult female distribution on the whole body surface of the host Lepomis gibbosus.

Fig. 4 - Lernaea cyprinacea: adult female distribution on the head region of the host Lepomis gibbosus. cyprinacea penetration were reported in literature (Berry et al., 1991; Silva-Souza et al., 2000; Hassan et al., 2008; Bednarska et al., 2009; Plaul et al., 2010). We also analysed, from the ecological point of view, the possible link between the size of the host fish and the parasite number. Infestation intensity with L. cyprinacea positively correlates both with the total length of the host bodies (r=0.192) and with their standard length (r=0.188). However, the null hypothesis that there is no significant relationship between the two variables could not be rejected, as the correlation coefficient (r) value was lower than the tabular value, at the chosen significance level (α = 0.05). The same positive but not significant correlations between host size and fish infections were also reported by Pérez-Bote (2000) and Gutiérrez-Galindo & Lacasa-Millán (2005) cited by Pérez-Bote (2010). The sampling season may interfere with the estimation of the parasite abundance and prevalence, as the water temperature is related with the reproduction of L. cyprinacea. It is well known that the period from egg to mature adult copepod INFESTATION OF LEPOMIS GIBBOSUS BY LERNAEA CYPRINACEA 67 is 13–14 days at 22–25°C (Al-Hamed & Hermiz, 1973). The life cycle of Lernaea species is completed in fewer days as the water temperature rises (Pérez-Bote, 2010). According to Mancini et al. (2006) cited by Pérez-Bote (2010), infestation parameters as prevalence and abundance of this parasite has a higher seasonal occurrence during warm months (June in our report), which coincides with blooms of bacteria, which in turns provides food for L. cyprinacea. In this study the copepod Lernaea cyprinacea was found infesting a sample of 80 Pumpkinseed sunfish. Biological material originates in the Moara Domneascã Lake, a sufficiently investigated area from the fauna diversity point of view, an ecosystem of the lentic type. The fixed parasites, especially in the head area, but also at the level of the fins and tegument, left their hosts with obvious signs of ulcerations and necroses. This is the first study regarding the distribution of the parasitic Lernaea cyprinacea on the surface of the body of Lepomis gibbosus in Romania.

ACKNOWLEDGEMENTS This study was supported by the PN II Research Program SAFAR no. 051-088/2007 allotted to Dr. Viorica Bãlan.

INFESTAREA BIBANULUI-SOARE LEPOMIS GIBBOSUS (TELEOSTEI: CYPRINIDAE) CU COPEPODUL LERNAEA CYPRINACEA (CRUSTACEA) – CÂTEVA ASPECTE ECOLOGICE

REZUMAT Copepodul Lernaea cyprinacea este cunoscut pentru spectrul mare de gazde piscicole pe care le infesteazã ºi la care poate produce hemoragii ºi ulceraþii. Lucrarea de faþã se concentreazã asupra unor aspecte ale relaþiei dintre bibanul-soare ºi acest copepod: parametrii infestãrii, situsurile de ataºare preferate, relaþia dintre dimensiunile gazdei ºi gradul de parazitare, precum ºi efectele vizibile ale acestui parazit lãsate pe suprafaþa corpului peºtelui.

LITERATURE CITED AL-HAMED, M., L. HERMIZ, 1973 - Experiments on the control of anchor worm (Lernaea cyprinacea). Aquaculture, 2: 45-51. BAUER, O. N., 1987 - Opredelitel’ parazitov presnovodnych ryb fauny SSSR. T. 3. (Guide to the parasites of the freshwater fish fauna of the USSR. Vol. 3. Parasitic Metazoa, Second Part). Nauka, Leningradskoe Otdelenie, 583 pp. (in Russian) BEDNARSKA, M., M. BEDNARSKI, Z. SO£TYSIAK, R. POLECHOÑSKI, 2009 - Invasion of Lernaea cyprinacea in rainbow trout (Oncorhynchus mykiss). Acta Scientiarum Polonorum, Medicina Veterinaria, 8 (4): 27-32. BERRY, C. R., Jr., G. J. BABEY, T. SHRADER, 1991 - Effect of Lernaea cyprinacea (Crustacea: Copepoda) on stocked rainbow trout (Oncorhynchus mykiss). Journal of Wildlife Diseases, 27 (2): 206-213. BOZ, E., V. VULPE, 1998 - Observaþii privind parazitismul cu Lernaea cyprinacea la peºtii de culturã. Sesiunea ºtiinþificã anualã a Facultãþii de Medicinã Veterinarã, Iaºi. (in Romanian) BUSH, A. O., K. D. LAFFERTY, J. M. LOTZ, A. W. SHOSTACK, 1997 - Parasitology meets ecology on its own terms: Margolis et al. revisited. The Journal of Parasitology, 83 (4): 575- 583. GAVRILOAIE, I. C., I. E. CHIª, 2007 - Consideraþii despre originea, rãspândirea ºi situaþia actualã a speciei Lepomis gibbosus (Linnaeus, 1758) (Pisces, Centrarchidae) în Europa ºi în România. Brukenthal. Acta Musei, Sibiu, 2 (3): 109-118. (in Romanian) HANGAN, M., L. D. URDEª, C. DIACONESCU, V. PETRICÃ, 2008 - Efectul patogenic al crustaceelor (Lernaea sp.) asupra peºtilor dulcicoli crescuþi în bazine statice, în unele amenajãri piscicole din zona Deltei Dunãrii. Lucrãri ªtiinþifice, Seria Zootehnie, USAMV Iaºi, vol. 51. (in Romanian) 68 MALA-MARIA STAVRESCU-BEDIVAN, OANA P. POPA, FLORIN T. AIOANEI, LUIS O. POPA

HASSAN, M. H., S. J. BEATTY, D. L. MORGAN, R. G. DOUPE, A. J. LYMBERY, 2008 - An introduced parasite, Lernaea cyprinacea L., found on native freshwater fishes in the south west of Western Australia. Journal of the Royal Society of Western Australia, 91: 149-153. IORGU, E. I., O. P. POPA, C. ADAM, C. PÂRVU, C. BAN-CALEFARIU, D. RUªTI, M. STAN, F. AIOANEI, M. STAVRESCU, V. TUDOR, V. BÃLAN, L. O. POPA, 2009 - Agroecosistems fauna diversity and their role in bioconservation – a case study: Moara Domneascã experimental centre. P. 87. In: D. Murariu, C. Adam, G. Chiºamera, E. Iorgu, L. O. Popa, O. P. Popa, (eds), Annual Zoological Congress of „Grigore Antipa” Museum – Book of abstracts, „Grigore Antipa” National Museum of Natural History, Bucharest. PATRICHE, T., N. PATRICHE, M. TENCIU, 2009 - Cyprinids total blood proteins determination. Lucrãri ºtiinþifice, Zootehnie ºi Biotehnologii, Timiºoara, 42 (2): 95-101. PÉREZ-BOTE, J. L., 2010 - Barbus comizo infestation by Lernaea cyprinacea (Crustacea: Copepoda) in the Guadiana River, southwestern Spain. Journal of Applied Ichthyology, 26 (4): 592-595. PLAUL, S. E., N. G. ROMERO, C. G. BARBEITO, 2010 - Distribution of the exotic parasite, Lernaea cyprinacea (Copepoda, Lernaeidae) in Argentina. Bulletin of the European Association of Fish Pathologists, 30 (2): 65-73. SILVA-SOUZA, A. T., S. C. ALMEIDA, P. M. MACHADO, 2000 - Effect of the infestation by Lernaea cyprinacea Linnaeus, 1758 (Copepoda, Lernaeidae) on the leucocytes of Schizodon intermedius Garavello & Britski, 1990 (Osteichthyes, Anostomidae). Revista Brasileira de Biologia, 60 (2): 217-220. VULPE, V., O. Z. OPREAN, S. PAªCA, 2001 - Infestaþii cu Lernaea cyprinacea la crap - etiopatogenezã ºi leziuni. Lucrãri ºtiinþifice, Medicinã Veterinarã, USAMV Iaºi, vol. 44. (in Romanian) TIRMIZI, H., 2003 - "Lernaea cyprinacea" (On-line), Diversity Web. Accessed August 9, 2010. http://animaldiversity.ummz.umich.edu/site/accounts/information/Lernaea_cyprinacea .html. Received: September 3, 2010 Mala-M. Stavrescu-Bedivan Accepted: May 09, 2011 University of Agronomic Sciences and Veterinary Medicine, Bdv. Mãrãºti No. 59, 011464 Bucharest, Romania e-mail: [email protected] Oana P. Popa, Luis O. Popa "Grigore Antipa" National Museum of Natural History ªos. Kiseleff No. 1, 011341 Bucharest, Romania e-mails: [email protected] [email protected] Florin. T. Aioanei University of Bucharest, Splaiul Independeþei No. 91-95, 050095 Bucharest, Romania e-mail: [email protected]