ANNALES · Ser. hist. nat. · 21 · 2011 · 2 119 Short scientific article UDK [593.191+595.74]:591.557.8 Received: 2011-09-13 EUGREGARINORIDA IN MYRMELEON AND EUROLEON: FIRST EVIDENCE OF THE GREGARINES (PROTOZOA: APICOMPLEXA) IN ADULT ANTLIONS (INSECTA: NEUROPTERA: MYRMELEONTIDAE) Dušan DEVETAK & Vesna KLOKOČOVNIK Department of Biology, Faculty of Natural Sciences and Mathematics, University of Maribor, SI-2000 Maribor, Koroška cesta 160, Slovenia E-mail: dusan.devetak@guest.arnes.si ABSTRACT During study of morphology of Neuroptera septate gregarines were found in the digestive tract of adult specimens of two antlion species, Myrmeleon hyalinus and Euroleon nostras. Micrographs and morphometric data of gamonts and trophozoites indicate that they belong to the genus Gregarina. However, additional studies will be required to identify gregarines at the species level. The present paper represents the first record of gregarines from adult antlions. Key words: Gregarines, Eugregarinorida, Neuroptera, antlions, Myrmeleontidae, parasites EUGREGARINORIDA IN MYRMELEON E EUROLEON: PRIMA EVIDENZA DI GREGARINE (PROTOZOA: APICOMPLEXA) IN ADULTI DI FORMICALEONE (INSECTA: NEUROPTERA: MYRMELEONTIDAE) SINTESI Durante uno studio sulla morfologia di neurotteri Formicaleone, sono state trovate gregarine nel tratto digerente di individui adulti di due specie, Myrmeleon hyalinus e Euroleon nostras. Dai dati morfometrici e dalle micrografie di gamonti e trofozoiti emerge che essi appartengono al genere Gregarina. Gli autori rilevano la necessità di studi aggiuntivi al fine di identificare le gregarine a livello di specie. L’articolo rappresenta la prima segnalazione della presenza di gregarine in individui adulti di Formicaleone. Parole chiave: Gregarine, Eugregarinorida, Neuroptera, Formicaleone, Myrmeleontidae, parassiti ANNALES · Ser. hist. nat. · 21 · 2011 · 2 120 INTRODUCTION Gregarines (Eugregarinorida) are relatively large pro- tozoan parasites in the guts and body cavities of several kinds of invertebrates, including annelids, tunicates, si- punculids and especially arthropods (Clopton, 2002; Ru- eckert & Leander, 2008). The anterior end is attached to the host via mucron in aseptate gregarines or epimerite in septate gregarines. Characteristic for gregarines is syzygy – the process in which two mature trophozoites pair up before the formation of a gametocyst. Life-cycle of nearly all species requires only one host. Gregarines move by gliding, bending and peristalsis (Clopton, 2002). The majority of eugregarine species are reported from insects. The knowledge of their occurrence in insects is poor; gregarines have been reported from less than one percent of named insect species (Clopton, 2002). The first gregarine genus recognized, Gregarina Dofour, 1828, was established for Gregarina ovata (Dofour, 1828) known as a parasite in the digestive trac- ts of earwigs (Dermaptera). In the period since Grega- rina was erected the genus has grown to include more than 300 species primarily infecting coleopterans and orthopterans. As is often the case with nominate gene- ra, Gregarina has become an agglomeration of taxa that includes a number of unrecognized or cryptic genera (see Clopton, 2002; Hays et al., 2004; Clopton & Hays, 2006; Clopton et al., 2008a, b). The first record of gregarines in lacewings (Neurop- tera) dates back to 1969 when Geus (1969) described Hyalospora hemerobii from brown lacewing Hemerobi- us pini. Later, in 1978 a new gregarine species, Actino- cephalus acanthaclisis was described from larval antlion Acanthaclisis baetica originating from France (Marques & Ormières, 1978). In the present paper, gregarines from adult antlions are reported for the first time. MATERIAL AND METHODS Adult antlions Myrmeleon hyalinus hyalinus (Olivi- er, 1811) and Euroleon nostras (Geoffroy in Fourcroy, 1785) were reared from larval stages. Larvae of M. hyali- nus hyalinus were collected in Douz (Tunisia) and larvae of E. nostras were collected in Katlanovo (Macedonia). Adults were fed in captivity with apricot jam and me- alworm larvae, Tenebrio molitor. The mealworm larvae were macerated before being consumed by the adults. Each antlion was dissected and its intestine was exa- mined microscopically at magnifications 100x, 200x and 400x. In total, 5 adults of M. hyalinus hyalinus and 3 adults of E. nostras were examined for gregarine pre- sence. Observed gregarines were measured and photo- graphed using a Nikon E 800 Microscope with a moun- ted digital camera Nikon DN100, and processed with Eclipse Net version 1.16.3 software. The following stan- dard parameters of gregarine body (in µm), according to Lipa (1967) and Clopton (2004), were measured to describe the characteristics of pathogens in the investi- gated antlions: total length, length of protomerite, length of deuromerite, width of protomerite, and width of de- utomerite. RESULTS Gregarines in Myrmeleon hyalinus hyalinus In the gut of one male of M. hyalinus hyalinus five gregarines were found. Individuals were elongated and solitary. Measurements are given in Table 1. Two morphological types were distinguished. In one gregari- ne type (Type A), measuring 122.9–152.9 µm in length, protomerite had hemispherical shape and deutomerite Tab. 1: Measurements of gregarines in Myrmeleon hyalinus hyalinus (in µm). Legend: TL - total length; LP - length of protomerite; WP - width of protomerite; LD - length of deutomerite; WD - width of deutomerite; WP:LP - ratio of the width of protomerite to the length of protomerite; WD:LD - ratio of the width of deutomerite to the length of deutomerite; WP:WD - ratio of the width of protomerite to the width of deutomerite; WP:TL - ratio of the width of protomerite to the total length; LP:TL - ratio of the length of protomerite to the total length. Tab. 1: Meritve gregarin v volkcih vrste Myrmeleon hyalinus hyalinus (v µm). Legenda: TL - celotna dolžina; LP - dol- žina protomerita; WP - širina protomerita; LD - dolžina dentomerita; WD - širina dentomerita; WP:LP - razmerje med širino protomerita in dolžino protomerita; WD:LD - razmerje med širino dentomerita in dolžino dentomerita; WP:WD - razmerje med širino protomerita in širino dentomerita; WP:TL - razmerje med širino protomerita in ce- lotno dolžino; LP:TL - razmerje med dolžino protomerita in celotno dolžino. Gregarines Parameter TL LP WP LD WD WP:LP WD:LD WP:WD WP:TL LP:TL Type A1 152.9 26.7 33 126.2 49.5 1.24 0.39 0.67 0.22 0.17 Type A2 128.5 24.6 25.1 103.9 42.3 1.02 0.41 0.59 0.20 0.19 Type A3 122.9 23.8 27.6 99.1 38.3 1.16 0.39 0.72 0.22 0.19 Type A4 143.8 24.4 32.3 119.4 45.5 1.32 0.38 0.71 0.22 0.17 Type B1 101.5 20.5 17.8 81 38 0.87 0.47 0.68 0.18 0.20 Dušan DEVETAK & Vesna KLOKOČOVNIK: EUGREGARINORIDA IN MYRMELEON AND EUROLEON: FIRST EVIDENCE OF THE GREGARINES ..., 119–124 ANNALES · Ser. hist. nat. · 21 · 2011 · 2 121 Gregarines Parameter TL LP WP LD WD WP:LP WD:LD WP:WD WP:TL LP:TL 1* 198 29 32.1 169 51.9 1.11 0.31 0.62 0.16 0.15 2* 160.2 24.6 34.9 135.6 46.8 1.42 0.35 0.75 0.22 0.15 3 239.4 34.6 33.3 204.8 52.8 0.96 0.26 0.63 0.14 0.14 4* 183.7 31.4 35.5 152.3 53.6 1.13 0.35 0.66 0.19 0.17 5* 151.1 24.3 28.3 126.8 42.9 1.16 0.34 0.66 0.19 0.16 6 212.5 30.4 33.8 182.1 54.5 1.11 0.30 0.62 0.16 0.14 7* 315 45.4 47.4 269.6 95.3 1.04 0.35 0.50 0.15 0.14 8* 372.1 27.2 50.3 344.9 115.1 1.85 0.33 0.44 0.14 0.07 9 201 27.9 31.3 173.1 68.3 1.12 0.39 0.46 0.16 0.14 10* 233.8 36.2 32.8 197.6 53.7 0.91 0.27 0.61 0.14 0.15 Tab. 2: Measurements of gregarines in Euroleon nostras (in µm). For legend see Table 1. Individuals marked with asterisk (*) were measured in syzygium. Tab. 2: Meritve gregarin v volkcih vrste Euroleon nostras (v µm). Za legendo glej Tabelo 1. Posamezni primerki, označeni z zvezdico (*), so bili izmerjeni v sizigiji. Plate 1 / Tabla 1: Figs. 1-3: Gregarines of the Type A in Myrmeleon hyalinus hyalinus. Figs. 1, 2: Individuals from the gut lumen. Fig. 3: Detail of the protomerite. Fig. 4: An individual of the Type B in M. hyalinus hyalinus. Sl. 1-3: Gregarine tipa A v volkcu vrste Myrmeleon hyalinus hyalinus. Sl. 1, 2: Osebki iz črevesnega lumna. Sl. 3: Detajl protomerita. Sl. 4: Osebek tipa B v volkcu vrste M. hyalinus hyalinus. Dušan DEVETAK & Vesna KLOKOČOVNIK: EUGREGARINORIDA IN MYRMELEON AND EUROLEON: FIRST EVIDENCE OF THE GREGARINES ..., 119–124 ANNALES · Ser. hist. nat. · 21 · 2011 · 2 122 was cylindrical to elongate ellipsoidal (Figs. 1-3). Con- striction was noted at protomerite deutomerite septum. The other type (Type B) with 101.5 µm length was bot- tle-like shaped and protomerite was broadly conical (Fig. 4). Gregarines in Euroleon nostras One female E. nostras contained at least ten grega- rines (Figs. 5-8). A few examples of syzygy were obser- ved and they were of caudofrontal type (Fig. 8). Mea- surements are given in Table 2. Gregarines measuring 151.1–372.1 µm in length were elongate ellipsoidal to tongue-shaped with hemispherical protomerites. DISCUSSION Keys and micrographs of many European gregari- ne species are provided in monographs of Lipa (1967) and Geus (1969). Marques & Ormières (1978) reported occurrence of gregarines in larval antlions. In the pre- sent study, gregarines are reported for the first time in adult antlions. Morphometric analysis of the gregarines found in the digestive tract of adult antlions indicates that they be- long to the genus Gregarina. According to the morphol- ogy, they resemble both gregarine species commonly occurring in Tenebrio molitor larvae, namely Gregarina steini Berndt, 1902 and Gregarina cuneata Stein, 1848. However, additional study will be required to identify gregarines in antlions at the species level. The adult ant- lions could be infected with gregarines during feeding with mealworm larvae. ACKNOWLEDGEMENTS We would like to express our thanks to Dr Paweł Sienkiewicz (University of Poznań, Poland) and Prof Dr Jerzy J. Lipa (Institute of Plant Protection, Poznań, Po- Plate 2 / Tabla 2: Figs. 5-8: Gamonts in Euroleon nostras. Fig. 5: A group of gregarines; syzygy is shown on the left. Figs. 6, 7: Two solitary individuals. Fig. 8: Syzygy of caudofrontal type. Sl. 5-8: Gamonti v volkcu vrste Euroleon nostras. Sl. 5: Skupina gregarin: na levi je prikazana sizigija. Sl. 6, 7: Dva posamezna osebka. Sl. 8: Sizigija kavdofrontalnega tipa. Dušan DEVETAK & Vesna KLOKOČOVNIK: EUGREGARINORIDA IN MYRMELEON AND EUROLEON: FIRST EVIDENCE OF THE GREGARINES ..., 119–124 ANNALES · Ser. hist. nat. · 21 · 2011 · 2 123 land) for critical reading of the earlier version of the ma- nuscript and their help and determination. This research was supported by the Slovene Ministry of Higher Edu- REFERENCES Clopton, R. E. (2002): Phylum Apicomplexa Levine, 1970: Order Eugregarinorida Léger, 1900. In: Lee, J. J., G. Leedale, D. Patterson & P. C. Bradbury (eds.): Illus- trated Guide to the Protozoa, 2nd Ed. Society of Protozo- ologists, Lawrance, Kansas, pp. 205–288. Clopton, R. E. (2004): Standard nomenclature and metrics of plane shapes for use in gregaine taxonomy. Comp. Parasitol., 71, 130-140. Clopton, R. E. & J. J. Hays (2006): Revision of the genus Protomagalhaensia and description of Protomag- alhaensia wolfi n. comb. (Apicomplexa: Eugregarinida: Hirmocystidae) and Leidyana haasi n. comb. 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Gustav Fischer Verlag, Jena, pp. 1–608. Dušan DEVETAK & Vesna KLOKOČOVNIK: EUGREGARINORIDA IN MYRMELEON AND EUROLEON: FIRST EVIDENCE OF THE GREGARINES ..., 119–124 cation, Science and Technology within the Biodiversity Research Programme (Grant No. P1-0078). EUGREGARINORIDA V VOLKCIH MYRMELEON IN EUROLEON: PRVA NAJDBA GREGARIN (PROTOZOA: APICOMPLEXA) V ODRASLIH VOLKCIH (INSECTA: NEUROPTERA: MYRMELEONTIDAE) Dušan DEVETAK & Vesna KLOKOČOVNIK Oddelek za biologijo, Fakulteta za naravoslovje in matematiko, Univerza v Mariboru, SI-2000 Maribor, Koroška cesta 160 E-mail: dusan.devetak@guest.arnes.si POVZETEK Med favnističnimi raziskavami mrežekrilcev smo v prebavilih odraslih volkcev dveh vrst, Myrmeleon hyalinus in Euroleon nostras, našli septatne gregarine. Na osnovi mikrografij gamontov in trofozoitov ter morfometričnih podat- kov sklepamo, da gregarine spadajo v rod Gregarina. Za določitev vrste bo potrebno opraviti še več opazovanj. 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In: The Tree of Life Web Project (http://tolweb.org/) Dušan DEVETAK & Vesna KLOKOČOVNIK: EUGREGARINORIDA IN MYRMELEON AND EUROLEON: FIRST EVIDENCE OF THE GREGARINES ..., 119–124