J Arthropod-Borne Dis, June 2013, 7(1): 94–98 M Badakhshan et al.: The First Report of… http://jad.tums.ac.ir Published Online: April 10, 2013 Case Report The First Report of Eustigmaeus johnstoni (Acari: Stigmaeidae) Parasitic Mite of Phlebotominae Sand Flies from Iran Mehdi Badakhshan 1, Javid Sadraei 1, *Vahideh Moin-Vaziri 2 1Department of Medical Parasitology and Entomology, College of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2Department of Parasitology and Mycology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran (Received 10 May 2011; accepted 1 Nov 2011) Abstract Background: Stigmaeids mites have been recorded only on Phlebotominae sand flies up to now. Five species of Eustigmaeus, and three of Stigmaeus were reported on infested sandflies in different country up to the present. Methods: Sand flies collection was done using CDC light trap and sticky paper. The mites were isolated from in- fested specimens, mounted in Puri’s medium and identified using reliable keys. Results: A mite infested Phlebotomus papatasi was observed during a study on sandflies of one of the southern provinces of Iran, near to the Persian Gulf. Several scars resulting from mite attachment were found on abdominal tergites of this female sand fly. The mites were identified as Eustigmaeus johnstoni. Conclusion: This parasitic mite is one of the eyeless species, which has a great distribution over the world, reported from Tunisia, Saudi Arabia, Pakistan, Yemen, Cyprus and Palestine. But, this is the first record of this species from Iran. Keywords: Eustigmaeus johnstoni, Phlebotomus papatasi, sand flies, Mite, Iran Introduction Five species of Eustigmaeus and three spe- cies of Stigmaeus (both Stigmaeidae), also one species of Dasythyreus (Dasythyreidae) have been reported as parasitic mites on insects yet. Stigmaeids mites have been recorded only on sand flies Eustigmaeus dyemkoumai (Abbonec 1970), E. gamma, E. gorgasi and E. parasitica (Chaudri 1965), E. johnstoni (Zhang and Ger- son 1995), (Shehata and Baker 1996), Stig- maeus smithi (Mitra and Mitra 1953), S. sinaei (Swift 1987) and S. youngi (Hirst 1926 and Wood 1972) were reported as parasitic mites of different species of Phlebotominae sand flies comprising Phlebotomus and Sergentomyia spp. Mites of genus Eustigmaeus Berlese, 1910 (Prostigmata: Stigmaeidae) comprise a group of globate, red acarines whose their bodies are covered by ornamental armour. In females this dorsal armour is separated into propodosomal and hysterosomal plates, the latter being often subdivided in the males. Females carry thir- teen pairs variously-shaped dorsal setae, of which three pairs are often ventrally displayed: the humerals (c2) laterally and the posterior- most b1 and b2 caudally. Most species have one pair of eyes, located between propodosomal setae ve and sci. The eyeless species include E. lirella, E. parasitica, E. gamma, E. gorgasi and E. johnstoni (Zhang and Gerson 1995). Parasitic mite, E. johnstoni has a broad dis- tribution and was reported from different countries including Yemen, Saudi Arabia, Pal- estine, Cyprus, Tunisia and Pakistan (Eddie et al. 2006). However, this is the first report of this *Corresponding author: Dr Vahideh Moin-Vaziri, E- mail: vmvaziri@gmail.com 94 J Arthropod-Borne Dis, June 2013, 7(1): 94–98 M Badakhshan et al.: The First Report of… http://jad.tums.ac.ir Published Online: April 10, 2013 mite from Iran. Several reports from different parts of the world provide this evidence that the Eustigmaeus-sand fly association is not accidental (Zhang and Gerson 1995). Materials and Methods Sand fly collection was done using CDC light trap and sticky trap in July 2010. We ob- served a sand fly (out of 286 collected speci- mens) which infested with mite, during a study on the Phlebotominae of Zirah Village (Bushehr Province) located south of Iran, in 2010. The mites were identified using the keys of Zhang and Gerson (1995) and Fan and Zhang (2005). Setal terminology follows Kethley (1990) and Zhang and Gerson (1995). Also some of the morphological characters were measured using gradient lens. All meas- urements are in micron. Accordind to Fan and Zang 2005 the fol- lowing characters were used to identify the Eustigmaeus spp from the other genus of Stig- maeidae (Key to genus Eustigmaeus, also key to eyeless Eustigmaeus species are men- tioned at the end of the article). Palptibial claw prominent, sub equal to palptarsus; with 2 pairs of subcapitular setae, Setae e1 and f1 situated on same shield in female (except Villersiella), without genital se- tae, Chelicerae separate, Prodorsal and dor- sal hysterosomal shields separate, Setae sce situated on main prodorsal shield and Setae d1 and e1 situated on same shield. Results The infested sand fly was identified as Phlebotomus papatasi (female). At least three mites (also female) were observed on the par- asitized specimen, one near to the legs and two others attached on the abdomen (Fig. 1a and 1b). Several scars were observed result- ing from mite attachment. The scars oc- curred only on the abdomen, particularly the anterior tergites. They had irregular outline, dark in color with a pale border (Fig. 2). According to the keys, the mite, was identified as Eustigmaeus johnstoni Zhang and Gerson, 1995 (Acai: Stigmaeidae) (Fig. 3a, 3b, 3c, 3d and 3e). According to Zhang and Gerson (1995) variation in the lengths and distances between dorsal setae are evi- dent in specimens from different countries. As Table 1 shows our measurements are comparable with the extent that he was stated, but due to the small number of mite in our access, the statistical comparison was not possible and meaningful. Fig. 1. Eustigmaeus johnstoni, located near the legs of infested Phlebotomus papatasi (a) and two others attached to its abdomen (b) Iran, 2010 95 J Arthropod-Borne Dis, June 2013, 7(1): 94–98 M Badakhshan et al.: The First Report of… http://jad.tums.ac.ir Published Online: April 10, 2013 Table 1. Measurements (µ ) of dorsal setae and setal distances in Eustigmaeus johnstoni (female), Iran, 2010 in comparison with other countries (Zhang et al. 1995) Yemen Saudi Arabia Cyprus Tunis Pakistan Iran n vi 22–32 19-23 24 21 21 25 1 ve 23-28 19-21 23 22 21 27 14 sci 18-23 15-20 21 21 19 20 15 sce 22-28 19-23 21 21 21-23 25 13 c1 22-26 19-23 21 23 21 27 14 c2 22-28 19-23 22 23 22-23 18 15 d1 19-28 19-21 21 23 21-22 26 14 d2 19-26 17-21 21 21 20-21 22 15 e1 22-28 21-23 23 21 20-21 23 14 e2 20-23 19-23 23 19 20-21 22 15 f 24-35 21-25 28 24 23-24 28 14 b1 22-32 20-26 24 21 24 24 15 b2 23-24 19-23 21 19 21 25 13 vi-vi 32-34 23-32 30 28 26-27 31 13 ve-ve 54-56 41-49 47 47 42-50 58 13 sci-sci 91-95 80-86 86 86 78-81 100 13 sce-sce 112-120 100-114 109 103 105 130 13 c1-c1 51-60 41-54 50 50 52-53 54 12 d1-d1 40-56 43-52 54 53 47-52 57 13 d2-d2 133-140 106-128 126 116 116 135 13 e1-e1 44-49 34-45 43 42 40 48 12 e2-e2 110-116 84-111 100 92 90 120 12 f-f 56-59 48-60 54 50 52-57 65 11 b1-b1 24-26 19-25 23 22 17-21 25 12 b2-b2 56-71 49-63 58 54 54 70 12 Fig. 2. Abdominal scars left on female Phlebotomus papatasi which was infested by three Eustigmaeus johnstoni mite, Iran, 2010 Fig. 3. Parasitic mite, Eustigmaeus johnstoni, on Phlebotomus papatasi from Iran, 2010, a. Idiosoma, dorsal view, b. Idiosoma, ventral view, c. upper part of Idiosoma, palp and legs, d. Chelicera, e. setae with long branches 96 J Arthropod-Borne Dis, June 2013, 7(1): 94–98 M Badakhshan et al.: The First Report of… http://jad.tums.ac.ir Published Online: April 10, 2013 Discussion Parasitic mite, E. johnstoni has a broad distribution and was reported from different countries, however this is the first report of this parasitic mite from Iran. Members of Stigmaeidae are soil or plant dwelling mites which are regarded as predators of other small arthropods (Swift 1987). Little is known about the relationship between mites and sand flies, whether this association is phoretic or parasitism? But several reports of scars, left by mites on infested sand flies (Mitra and Marta 1953, Abbonec 1970, Lewis and Macfarlane 1982) strengthen the parasitic relationship. However, the nature of Eustigmaeus-sand fly association remains indeterminate. There is no report of males or immature of Eustigmaeus on phlebotominae sandflies. It could be postulated that development and mating of the mentioned mite were done else- where, probably in the habitat where sandflies breed and rest. So, additional work at differ- ent times of the year also in other areas, with investigations of sand fly breeding and rest- ing sites are essential to provide further in- sight to the significance of the mite-sand fly association. Also the exact nature of these parasitic mites infestation is poorly under- stood and further works are required to de- termine if such heavily infected sand flies have reduced longevity. Key to eyeless Eustigmaeus Genus (Zhang and Gerson 1995) 1- Dorsal hysterosomal setae c1 and d1 short, not reaching the bases (alveoli) of any neighboring setae...................................... 2 - Dorsal hysterosomal setae c1 and d1 long, over reaching the bases (alveoli) of at least two neighbouring setae........................... 4 2- Distance between alveoli of dorsal setae pairs, c1-c1, d1-d1 and e1-e1 subequal........... 3 - Distance c1-c1 and d1-d1 subequal, both mach less than e1-e1 (setae e1 laterally dis- placed) ………….…… Eustigmaeus gamma 3- Dorsal setae flat, with short barbs on dis- tal ¾; all intercoxal setae (1a, 3a and 4a) weakly barbed…………...……….. E. lirella - Dorsal setae thin, with long branches throughout; intercoxal setae (1a, 3a, and 4a) with long branches………….... E. johnstoni Acknowledgements The authors wish to thank Dr Zhi-Qiang Zhang and Prof. Leger for their kind collab- oration in providing some key references. This study was financially supported by the Department of Medical Parasitology and En- tomology, College of Medical Sciences, Tarbiat Modares University, also Department of Med- ical Parasitology and Entomology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences. The authors declare that there is no conflict of interests. References Abbonec E (1970) Notes sur les Acarines parasites des Phlebotomus. Cahiers L’office de la Recherche Scientifique et Teqnique Oter-Mer. Entomol Med Parasit. 8: 89–94. Berlese A (1910) Acari nuovi. Manipulus V, VI. Redia 6: 199–234 + Plates XVIII– XXI. Chaudhri WM (1965) New mites of the genus Ledermuelleria. Acarol. 7: 467–486. Eddie AU, Harten A, Magdalena KP (2006) The mites and ticks (Acari) of Yemen: an annotated check-list. Fauna of Ara- bia. 22: 243–286. Fan Q, Zhang ZQ (2005) Fauna of New Zea- land. 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