MORPHOLOGICAL IDENTIFICATION OF SCORPION SPECIES FROM JAZAN AND AL-MEDINA AL-MUNAWARA REGIONS, SAUDI ARABIA

The present work is a complementary contribution to the comprehensive study of the scorpion sting syndrome in Saudi Arabia. It deals with the identification and determination of medically important scorpions and the other ones, which were collected from two regions (Jazan and Al-Medina Al-Munawara), based on their morphology (the molecular phylogeny and venom characteristics will appear in subsequent publications). The specimens collected from those two regions were brought to the Research Center laboratories in several batches. Morphological identification of the collected specimens was done employing identification keys. There were 646 specimens collected from Jazan Region. A single species, Nebo hierichonticus (Family Diplocentridae), and five genera with four identified species, Parabuthus liosoma, Hottentotta jayakari (salei?), Compsobuthus werneri, Leiurus quinquestriatus (Vachoniolus globimanus?), Vachoniolus spp. (other species) and Orthochirus innesi (Family Buthidae), were classified as extant scorpions in the region. Three hundred and ninety-six specimens from Al-Medina Al-Munawara Region were categorized into eight groups; four of them were identified. Three buthids, Leiurus quinquestriatus, Androctonus crassicauda, Orthochirus innesi and one scorpion specimen, Scorpio maurus, were identified and classified as extant scorpions in this region. The other four species are still not completely identified. They are Vachoniolus (Buthacus minipectinibus?) globimanus? (Unidentified-1), Compsobuthus spp (arabicus?) (Unidentified-2), Compsobuthus spp (werneri?) (Unidentified-3) and a single specimen of Androctonus spp (australis?) (Unidentified4), all of which belong to the family Buthidae.

A decade ago, there were only nine recognized families (56) but the recent increase indicates a considerable activity in high-level (subfamilies and upper) scorpion taxonomy.The result was the discovery of entirely new families such as Pseudochactidae (19); elevating in rank existing subfamilies, such as Euscorpiinae, Superstitioniinae (62), Heteroscorpioninae (33), Hemiscorpiinae and Urodacinae (36,44), and creating new taxa at family rank for genera formerly placed in other families such as Microcharmidae and Troglotayosicidae (33)(34)(35).
Rank and single phyla of some newly created families lead to dispute (36,(44)(45)(46)(47) on family status of Hadogenidae and Lisposomidae.Relevant to the systematics of extant scorpion families is the recent progress in reassessing of existing fossil scorpion taxa and description of new ones.Kjellesvig-Waering (25) revised all fossil scorpions.The majority of which belong to now extinct lineages at the level of suborders and infraorders.This is why especially important are the recent discoveries of Cretaceous orthostern scorpions from Brazil (5,6), Lebanon (37), Burma (38,54), and France (39).
The studies of Prendini and Wheeler (48) summarized the works on classification of recent (extant) scorpions during the past 25 years.Scorpion families were seven (30), the number was raised to 13 (61), reduced to nine (43,56), reverted to 13 (27), again raised to 16 (14), further increased to 20 (36), then reduced to 14 (57), raised to 18 (48) and back to 13 (15, 58).These works were thoroughly analyzed and professionally criticized by Prendini and his colleague Wheeler (48), suggesting that a centralized record of taxa might be the only solution for ensuring quality control in the taxonomy and future publishing.Fet and Soleglad (15) have rejected this criticism and reverted to their previous classification.
Previous scorpion studies and surveys in Arabia (1,2,65) confirmed the presence of only two species (Androctonus crassicauda and Leiurus quinquestriatus) in Al-  (65).In fact, these two species of Al-Medina Al-Munawara Region are the most toxic and hence medically important species (24, 55).New synonyms were found in the taxonomy of some families of Old World scorpions ( 9), but Fet and Braunwalder (10) have tackled the preparation of accurate identification keys dealing with current problems in taxonomy and biogeography of the Eastern Mediterranean area.Some recent studies were provided concerning the scorpion fauna of the Middle East (66).
The family Buthidae, with 48 genera and over 500 species, is the largest and most widespread of the scorpion families (12,43).Its members occupy all six faunal regions, but the greatest generic diversity occurs in the Old World, especially in the Afrotropical Region (22 genera) and in southern portions of the Palaearctic Region (23 genera).Although there have been many attempts to recognize subfamilies in Buthidae, none appears satisfactory (60).However, the interested reader is referred to other sources of information (4,26,28,31,42,49,68), in addition to websites of scorpiology (3, 15, 22, 67).
The family Diplocentridae had been represented by a single species in Saudi Arabia so far, but this family has some features and relationships in common with the family Scorpionidae (43) and now is considered part of it, according to recent arguments of classification (48).
The family Scorpionidae includes seven genera distributed among three subfamilies.
Scorpionidae contains some of the world's largest living scorpions.Heterometrus (Gigantometrus) swammerdami reaches lengths up to 16.8cm (7) The aims of the present work are to confirm the already reported scorpions as well as to record the newly identified ones in Jazan and Al-Medina Al-Munawara Regions, Saudi Arabia.

Scorpions
Professional hunters, who were specifically recruited for this job, hunted down and collected the scorpions from the wild, infested areas of Jazan and Al-Medina Al-Munawara Regions.Jazan Region is on the eastern coast of the Red Sea in the southwest of Saudi Arabia and Al-Medina Al-Munawara Region is in the northwest, not far from the eastern coast of the Red Sea (Figure 1).Scorpion specimens were brought to the Riyadh Military Hospital (RMH), Research Center Laboratories in groups (batches).

Maintenance of Living Scorpions and Preservation of Died Scorpions
Living scorpions were kept in plastic boxes with water ad libitum and fed with mealworms every fortnight.Yellow mealworms were supplied from a local animal facility, raised with the beetle Tenebrio molitor Linnaeus (William, 40, 41).Died scorpions were preserved in 70-96% ethanol and stored in our laboratory for morphological identification.

Morphological Examination and Diagnosis of Scorpions
Sub-grouping and crude morphological identification (names) of well-known groups were done visually, based on experience.The full identification and classification attributes of the other unidentified scorpions were done by running the taxonomic keys of Vachon (65) and Polis (43), employing microscopy.More recent reviews and

Scorpions
All specimens of scorpions were morphologically divided into groups that resemble each other.Taxonomically, scorpions were categorized into the following higher classification: Phylum: Arthropoda Class: Arachnida Order: Scorpiones (Scorpionida)

Morphological Identification and Diagnosis of Scorpion Genera and Species according to Regions Region 1 (Jazan Region):
Five batches were collected from Jazan Region that gave a total count of 646 specimens of scorpions (Table 2).
Existence of the identified genera (Parabuthus, Hottentotta, Compsobuthus, Orthochirus and Nebo) was reconfirmed by verification of the keys of Sissom (56) and Vachon (65).Species of Nebo and Hottentotta were neither mentioned by Sissom nor by Vachon in these keys.Hence, these two genera were further comparatively verified with the fauna specimens and figures of Oman, which is bordering the region.Nebo hierichonticus and Hottentotta jayakari were recognized, although the latter species demands further verification to the subspecies (salei?).
The unidentified groups await further microscopic differentiation, but suggestive names were submitted.

Region 2 (Al-Medina Al-Munawara Region):
There were seven groups in the three collected batches from Al-Medina Al-Munawara Region, giving 396 scorpion specimens (Table 3).The existence of four groups was confirmed and they were identified to species according to the keys of Sissom and Vachon (56,65).
All these groups belonged to the representative medically important family Buthidae.
It is interesting to observe the situation about the combined and overlapped coexistence of species of these two families (Buthidae and Scorpionidae) with regard to their toxicity, according to previous reports (8,24,55).Region.Further collections and morphological assessments will be carried out in the next surveys and works to reconfirm the common scorpions endemic in these two regions and the bordering ones.

Figure 1
Figure 1 Map of the Kingdom of Saudi Arabia showing the two study areas of Jazan and Al-Medina Al-Munawara Regions (reformed from Saudi Maps).

Figure 3
Figure 3 Dissected parts of the pedipalp showing type A basic trichobothrial pattern.According to Sissom (54). A. dorsal view of femur.B. dorsal view of patella.C. external view of patella.D. ventral view of patella.E. external view of chela.F. internal view of chela.G. ventral view of chela.d: dorsal; i: internal; e: external; b: basal; sb: sub basal; m: medial; v: ventral; st: subterminal; t: terminal.
Androctonus crassicauda andLeiurus quinquestriatus, which are considered highly venomous(8), added to Androctonus spp.(australis?), which is considered the most important species in the Old World, are ones of the most toxic scorpions of the world compared with the relatively less toxic and smaller sized ones (22).Hence, new species are now recognized to be extant in the present surveyed regions, without any previous records, based on the available data submitted by the mentioned investigators.Chronic problems of scorpion classification are now the subject of DNA-based, modern molecular phylogenetic technique applications.These techniques are opening the way for identification methodologies that can deal with the scorpion fauna of the most complicated parts of the world, especially the Eastern Mediterranean and the Middle East(10, 18).Prendini and Wheeler(48) have analyzed recent works of the last three decades and proposed solutions for problems of scorpion higher taxonomy and online publishing concerning classification.The molecular data analyses that are combined with the detailed morphological investigations and exhaustive geographic sampling(10)(11)(12) 16) are dealt with elsewhere in future works.The preliminary morphological determination of the present survey confirmed the existence of eight species in Al-Medina Al-Munawara and seven species in Jazan

Table 1 .
(43)HOLOGICAL IDENTIFICATION OF SCORPION SPECIES FROM JAZAN AND AL-MEDINA AL-MUNAWARA REGIONS, SAUDI ARABIA.J. Venom.Anim.Toxins incl.Trop.Dis., 2007, 13, 4, p.  826online taxonomy were not fully implemented here, due to the ongoing works and arguments tracing this matter.Numbers, basic patterns and nomenclature of scorpion trichobothria and their carinae, indicating their locations on the surfaces of body and limbs.According to Polis(43).
(43,56,65)mari et al.keys of Vachon(65)deal with the same phylogenetical levels (identifying the species and subspecies), but limited to those extant in the Arabian Peninsula.These keys are also reliable in dealing with juvenile scorpions.The trichobothrial patterns(43,56,65)were the backbone in identification (Table1, Figures2 and 3), assisted by the employment of light (Stereo zoom dissection) microscopy.d: dorsal; i: internal; e: external; b: basal; sb: sub basal; m: medial; v: ventral; st: subterminal; t: terminal.