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A Newly Recorded Neogregarine (Protozoa, Apicomplexa), Parasite in Honey Bees (Apis Mellifera) and Bumble Bees (Bombus Spp)

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A Newly Recorded Neogregarine (Protozoa, Apicomplexa), Parasite in Honey Bees (Apis Mellifera) and Bumble Bees (Bombus Spp) Note A newly recorded neogregarine (Protozoa, Apicomplexa), parasite in honey bees (Apis mellifera) and bumble bees (Bombus spp) JJ Lipa 1 O Triggiani 1 Institute of Plant Protection, Miczurina 20, 60-318 Poznan, Poland; 2 Istituto di Entomologia Agraria, Universita degli Studi, via Amendola 165/A, 70126 Bari, Italy (Received 10 February 1992; accepted 31 August 1992) Summary — A new infectious disease of Apis mellifera, Bombus hortorum and B terrestris is re- ported to be caused by a parasitic protozoan belonging to the order Neogregarinida. The disease can be diagnosed based on the presence of characteristic navicular spores measuring 11.4-14.4 x 3.6-5.4 μm. Infection level in host populations was low but since this parasite was found in host in- sects in Finland and Italy, it evidently has a wide distribution in Europe. Apis / Bombus / neogregarine / protozoan / parasite While continuing our studies (Lipa and pathogen was also recorded in 1990 in Triggiani, 1988; Triggiani, 1991) on spiro- one A mellifera worker from a hive in the plasma, flagellate and microsporidian in- experimental apiary at the Agricultural Re- fections of the honey bee (Apis mellifera search Center in Jokionen, Finland, it was L) and bumble bees (Bombus spp) in concluded that the pathogen was a new some adults collected in Italy and in Fin- parasitic protozoan which, based on the land very peculiar spores were observed type of spores and the life cycle, belongs which could not be attributed to known to the order Neogregarinida. of these beneficial insects. A pathogens The detailed data on host, country and is therefore preliminary report presented locality records in which the new pathogen to alert researchers in various countries was noted are given in table I. These data as to the occurrence and incidence of the indicate that the infection level by this neo- involved. pathogen gregarine is very low, which is not surpris- The new pathogen was first noticed in ing as infections of the honey bee (A mellif- 1988 in flying workers and queens of era) by eugregarines (Stejskal, 1965) or Bombus hortorum L and B terrestris L col- flagellates (Langridge, 1966) are also gen- lected by sweep net from various crops in erally at a very low level and have been re- Apuglia (southern Italy). When the same ported from very few localities. However, since this parasitic neogregarine was found in Finland and Italy we assume that it has a wide distribution in Europe and that a thor- ough survey may indicate that it also occurs in other countries or continents. The following diagnostic features may be helpful in survey and identification of the pathogen involved. On smeared fat body preparations of A mellifera and Bom- bus spp, various developmental stages were observed. Micronuclear meronts con- tain up to 40 nuclei and have a diameter of up to 45 μm (fig 1). Micronuclear mero- zoites are oval or elongated in shape and measure 5-9 x 2.5-3.5 μm (fig 1). Macronuclear meronts measure 20 x 15 μm and give rise to ovoid or pyriform mac- ronuclear merozoites 5-8 μm in diameter (figs 1,2). The spores observed in Apis and Bom- bus were identical. They are navicular in shape, and have distinct caps (figs 3,4). développement du parasite, étudié sur des Spores fixed in methanol for 2 min and frottis du corps gras fixés 2 min au métha- stained with 0.25% Giemsa for 15 h are nol et colorés avec une solution de Giem- 11.1—14.4 μm long and 3.6-5.4 μm wide. sa à 0,25% durant 8-12 h, le parasite ap- Inside the stained young spores 4 uninu- partient à l’ordre des Neogregarinida. Les cleate sporozoites are readily visible (fig mérontes macronucléaires renferment 3); however, they become less visible in jusqu’à 40 noyaux, leur diamètre atteint the mature spores (fig 4). 45 μm. Ils mesurent 20 x 15 μm et donnent naissance à des mérozoïtes macronucléai- The life cycle and morphology of me- res ovoïdes ou un ronts and spores indicate that the proto- (ma) pyriformes ayant diamètre de 5-8 Les méro- zoan infecting A mellifera and Bombus spp μm (figs 1-2). belongs to the order Neogregarinida (= zoïtes micronucléaires sont de forme ovale Schizogregarinida) (Apicomplexa). Its ou allongée et mesurent 5-9 x 2,5-3,5 μm observées chez et spores (sporocysts) differ greatly from (fig 1 ). Les spores Apis chez Bombus étaient Elles sont those of presently known genera as they identiques. naviculaires et des contain only 4 sporozoites while spores of possèdent chapeaux distincts aux 2 extrémités. Les the genera eg Mattesia, Farinocystis or spores Syncystis contain 8 sporozoites. In addi- fixées et colorées ont 11,1-14,4 μm de et de À l’intérieur tion, the spores of this newly reported neo- long 3,6-5,4 μm large. des colorées on facilement re- gregarine are narrow and have a delicate spores peut wall. All these features indicate that this connaître 4 sporozoïtes contenant chacun un La des neogregarine constitutes both a new spe- noyau (fig 3). morphologie pre- miers stades de et les ca- cies and a new genus. It also represents développement the first record of neogregarine infection in ractéristiques morphologiques des spores the Apidae. différencient ce nouveau néogrégarine des genres Mattesia et Farinocystis. Nous considérons donc que le néogregarine dé- ACKNOWLEDGMENTS couvert chez Apis et Bombus est un genre nouveau et une espèce nouvelle. Comme le montre le tableau I, le niveau The senior author his thanks to (JJL) expresses d’infestation dans les colonies d’abeilles et the National Research Council for Agriculture de bourdons étudiées était relativement and Forestry of the Academy of Finland for pro- viding funds to conduct a study in Finland faible. Néanmoins l’intensité de l’infestation (June—October, 1990) and to the University of chez certains individus parasités était très Bari for support in conducting this study in Italy forte. Ceci prouve que le pathogène peut (December 1990). diminuer les processus vitaux et les activi- tés des hôtes parasités. Sa répartition géo- graphique est étendue puisqu’on l’a trouvé Résumé — Un nouveau néo- parasite aussi bien en Finlande qu’en Italie. grégarine (Protozoa, Apicomplexa) de l’abeille (Apis mellifera) et du bourdon (Bombus sp) (Hymenoptera, Apidae). Zusammenfassung — Ein neu beschrie- L’examen au microscope de tissus benes neogregarines parasitisches Pro- d’adultes d’Apis mellifera, récoltés en 1990 tozoon bei Apis mellifera und Bombus en Finlande, et de Bombus terrestris et de sp. Die mikroskopische Untersuchungen B hortorum, récoltés en 1988 en Italie, a von 1990 in Finnland gesammelten A mel- montré la présence de spores d’un proto- lifera-Arbeiterinnen und von Bombus terre- zoaire encore inconnu. D’après le cycle de stris und B hortorum, die 1988 in Italien ge- sammelt worden waren, ergab den Befund Wie aus Tabelle I zu ersehen, ist der von Sporen eines bisher unbekannten pa- Infektionsgrad bei den untersuchten Bie- rasitischen Einzellers. Der Lebenszyklus nenvölkern und Hummelkolonien relativ des Parasiten, untersucht an Ausstrichen niedrig. Dennoch war die Intensität der In- des Fettkörpers (2 min fixiert in Methanol fektion bei einigen erkrankten Tieren ziem- und gefärbt in 0.25% Giemsa-Lösung für lich hoch und dies beweist, daß der Erre- 8-12 h) bewies, daß er zur Ordnung der ger die Lebensvorgänge und die Aktivitä- Neogregarinida gehört. Macronucleäre ten der befallenen Wirte beeinträchtigen Meronten enthalten bis zu 40 Nuclei und kann. Der Erreger hat offensichtlich eine besitzen einen Durchmesser von bis zu 45 weite geographische Verbreitung, da er μm. Micronucleäre Merozyten sind oval sowohl in Finland wie in Italien beobachtet oder länglich und messen 5-9 x 2.5-3.5 μm wurde. (Abb 1). Macronucleäre Meronten messen 20 x 15 μm und erzeugen ovoidale oder Parasitologie / Protozoa / Neogregarini- birnenförmige macronucleäre Merozyten da / Apis / Bombus (ma) mit einem Durchmesser von 5-8 μm (Abb 1-2). Die bei Apis und Bombus beobachteten REFERENCES Sporen (Oozysten) waren gleich. Sie haben die Form eines Bootes und tragen Langridge DP (1986) Flagellated protozoa (Fam an beiden Enden deutliche Kappen. Fixier- Trypanosomatidae) in the honey bee, Apis te und gefärbte Sporen sind 11.1-14.4 μm mellifera in Australia. J Invertebr Pathol 8, lang und 3.6-5.4 μm breit. Im Inneren der 124-126 gefärbten Sporen sind deutlich vier Sporo- Lipa JJ, Triggiani O (1988) Crithidia bombi sp n, zoiten mit je einem einzigen Kern zu a new flagellated parasite of a bumble bee sehen (Abb 3). Bombus terrestris L (Hymenoptera, Apidae). Acta Protozool 27, 287-290 Die Morphologie der frühen Entwick- M Acuta rousseaui n n lungsstadien und morphologische Merk- Stejskal (1965) sp, gen, n fam, eine neue cephaline Eugregarine, Par- male der unterscheiden diese neu Sporen asit der Honigbiene. Bull Apic 8, 217-225 beschriebene Protozoe von den Gattun- O Micro e endo- Mattesia und Deshalb Triggiani (1991) macroganismi gen Farinocystis. zoici in adulti di Bombus Latr e Psithyrus Lep betrachten wir die von uns bei Apis und (Hymenoptera: Apidae). Atti XVI Congr Nazi- Bombus neu beschriebene Neogregarine onale Italiano di Entomologia (Bari-Martina als neue Gattung und neue Art. Franca 23-28 September 1991) 587-597.
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