Asian Pacific Journal of Tropical Medicine (2010)800-803 800

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Document heading Geographical distribution and ecological features of the subspecies in the main zoonotic cutaneous leishmaniasis foci in Mohammad Reza Abai, Mohammad Ali Oshaghi*, Leila Tajedin, Yavar Rassi, Amir Ahmad Akhavan

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

ARTICLE INFO ABSTRACT

Article history: Objective: Rhombomy opimus (R. opimus) To reveal subspecies composition of in Iran. Received 12 July 2010 Methods: In this study, field specimens of the gerbil were collected from all its geographical Received in revised form 27 July 2010 Accepted 15 August 2010 rang in northeastResults: and central parts of Iran and identified on basisR. opimus of morphological sodalis R. characteristics. Results revealed presence of two subspecies of and Available online 20 October 2010 opimus sargadensis in the country. The first subspecies with brown to hazel-nut color and a size R. opimus Keywords: bigger than a typical was found only in Golestan province, an isolated colony which is Rhombomys opimus located in plains of north slopes R.of Alborzopimus Chainsargadensis Mountains of the country, connected with the . The subspecies of with a yellowish color and similar size as R. opimus Great gerbil a typical was found in all other areas of the great gerbil distribution in Iran. There was Ecology a distinct topographicConclusions: difference but similar ectoparasites between colonies of the two subspecies Reservoir in the study area. Further ecological and genetic investigations are required for Leishmaniasis R. opimus R. opimus- Phlebotomusmore detailed papatasi-Leishmania description of the major subspecies range and structure. The Iran association and the ZCL severity and outcome in hosts are discussed.

[2] 1. Introduction recorded from northern and western (Baluchistan)[11]. In the species is very widespread, [12] Rhombomys opimus (R. opimus) and has been recorded from many areas . In , , the great gerbil, is the it has been recorded from and semi-desert habitats main reservoir of zoonotic cutaneous leishmaniasis across southern Mongolia[13]. [1-3] R. opimus (ZCL) in Iran,Leishmania Afghanistan, major and . The rate of is a diurnal, fossorial, colonial often seen infection to , the cause of ZCL, is normally associated with other gerbils. The social structure consists [3-5] high and might raise more thanLeishmania 90% in Leishmaniaendemic areas gerbilli. of family groups that occupy isolated systems of (L.Additionally, gerbilli two Leishmaniaspecies of turanica ,(L. turanica) consisting of one breeding male, from one to seven females, ) and which and juveniles[14]. In China, the species occupies desert to are nonpathogenic to humans but can infect the great gerbil, semi-desert habitats, and is most successful in dry river [12] have been Leishmaniafound in Turkmenistan, Uzbekistan, China, and beds dominated with shrubby vegetation . In South Iran[1,3,6,7]. parasites persist in great gerbils for Asia, it has been recorded from apple orchards and clay- long time up to 25 months[1]. Great gerbils are also known sandy embankments and has been found to occupy Steppe to carry bubonic plague[8] as well as being an agricultural Mountains and upland and sand dunes with scattered pest[9]. vegetation[14]. It constructs large entrance holes to a very This species of gerbil is found in many parts of Central elaborate system that consists of long deep tunnels; Asia such as Turkmenistan and , Iran, Southern nest and food storage chambers[12]. Settlements of the Mongolia, North-western China and Afghanistan. It great gerbil are also associated with the borders of saline is widespread in Iran[3-5], Turkmenistan, Uzbekistan, depressions with plant communities dominated by annual Kazakhstan, and appears to have a smaller distribution in and perennial succulents in the family Chenopodiaceae Climacoptera Salsola Suaeda [14] Kyrgyzstan and Tajikistan[1,10]. In South Asia, it has been ( spp., spp., spp.) ’. Lifespan varies from male to female. The males maximum [15] lifespan is 2 to 3 years and the female lives 3 to 4 years . *Corresponding author: MA Oshaghi, Department of Medical Entomology and Vector R. opimus Control, School of Public Health, Tehran University of Medical Sciences, Tehran lives in large colonies composed of many 14155-6446, Iran. subgroups. These subgroups are believed to be offspring Tel: 0098 21 88951393 of mother/father pairs within the colony. During the winter Fax: 0098 21 88951393 these large groups huddle together to keep warm when E-mail: [email protected] Mohammad Reza Abai et al./Asian Pacific Journal of Tropical Medicine (2010)800-803 801 R. opimus opimus R. opimus sargadensis temperatures become critically low outside. They rely on the were considered as stable underground temperatures (20 to 25曟) which exist in subspecies (Figure 2B/C). their burrows. This species does not hibernate, although its activity is reduced during the winter months[9]. According to the classification of Etemad[16] and [17] Rhombomys opimusGoodwin opimus, four (R. known opimus subspecies opimus) Rhombomysnamed opimus fumicolor (R. opimus fumicolor), ,Rhombomys opimus sargadensis (R. opimus sargadensis) Rhombomys opimus sodalis (R. opimus sodalis) , and R. opimus opimushas been describedR. opimus in itsfurmicolor geographical rang. and were reported from central Asia, eastern region of Caspian Sea, including countries of Turkmenistan, [18] R. opimusUzbekistan,sargadensis KazakhstanR. opimus. Othersodalis two subspecies of and were reported in [16] R. opimus sodalis R. opimus opimus Iran . R. opimus is fumicolorbigger than but smaller than . This subspeciesR. opimus is sargadensisbrown to hazel-nut color. The subspecies of R. opimus has a yellowish color and similar size as a typical . In this paper the distribution of the great gerbil subspecies in Iran was analyzed on the basis of morphological characteristics of the field specimens collected from the Figure 1. R. opimus main areas of its habitats in the country constitute distinct Details of geographical locations where specimens populations. were collected in Iran. 1: Kalaleh (Golestan P.); 2: Damghan (Semnan P.); 3: Ahmad Abad, Shahrood (Semnan P.); 4: Mayamay, Shahrood (Semnan P.); 5: 2. Materials and Methods Esfarayen (Northern Khorasan P.); 6: Shirvan (Khorasan Shomali P.); 7: Badrood (Esfahan P.); and 8: Habibabad (Esfahan P.). 2.1. Sample collection

A Sample collection was performed in spring (April-June) 2008 in five provinces including Esfahan, Fars, Northern- Khorasan, Semnan, and Golestan Ps located in north, north- east, and central regions of the country (Figure 1). Colonies of the gerbils located about 1-1.5 km around the villages. All gerbil colonies, including occupied and unoccupied burrows were checked to select and to mark the active burrows. The active colonies of were identified® and rodents B were captured by setting the Sherman live traps (30 cm伊 C 15 cm伊15 cm wire mesh live-traps) baited with a mixture of cucumbers, tomatoes, and peanut butter. Ecological and biological data such as topographic features, plantations, daily activities, and abundance of colonies was recorded in each region. 2.2. Species and subspecies identification Figure 2. R. opimus R. Photos of two subspecies found in Iran. A/B: opimus sargadensis R. opimus sodalis and C: . The genus and species of the rodents were determined by external characteristics including color, body 3. Results measurements, ears, tail, feet, teeth, and skull[19]. According to classification of Etemad and Goodwin[16,17], the animal is 400 We caught 50 adult specimens including 31 females and one of the largest gerbil species up to mm from head to 19 tail. The upper body is yellowish-orange or dark grayish- males. Details of the captured specimens were shown in Table 1. The most active colonies were found in Habibabad, yellow, matching the sandy deserts they inhabit. Great (P.) gerbil is significantly bigger than most gerbils weighing Esfahan province . In this area, the burrows are dug in about 200 g, having a larger head and small ears than most plain area at sandy-loamy soil. The development of desert gerbils. It has a relatively short tail with a distinctive crest of control projects resulted in suitable’ habitats for increasing long hairs towards the end. Great gerbils have thick, dense, and flourishes of great gerbils population. In general, and soft fur and its body is stocky. The claws are long and observations on ecotype of this animal in the country showed sharp to help it dig its burrows[9]. In addition to the general that great gerbils preferred sandy soils, desert, and semi- characteristic mentioned above, presence of two fine grooves desert areas and subtropical steppes. Their diet mainly consisted of desert bushes and local agricultural crops. on theR. opimus incisor teeth is the most important characteristics for identification. The specimens with hazel-nut In hot seasons, they were more active at dawn and dusk, color, body length ranging from 300 to 320 mm and a tail whereas in winter, they were more active during the middle of the day. length of 135 to 140 R.mm, opimus and backsodalis feet lengths of 43-45 mm were considered as subspecies (Figure 2A). Large parts of the animal territory with a high number of Specimens with brown to yellowish color and similar size as populations were found both in the south (Esfahan P.) and Mohammad Reza Abai et al./Asian Pacific Journal of Tropical Medicine (2010)800-803 802 Table 1 R. opimus Details of subspecies specimens collected in different locations in Iran. Subspecies No. Male/female Province District Village R.opimus sodalis 5 2/3 Golestan Kalaleh Ghareh Gol Gharbi, Aziz-Abad, Allah nor R.opimus sargadensis 4 2/2 Northern- Khorasan Esfarayen Kalateh-Shor, Emarat, Toy R.opimus sargadensis 1 1/0 Northern- Khorasan Shirvan Hossein-abad, Shorak R.opimus sargadensis 5 2/3 Semnan Shahrood Ahmad-Abad, Mayamey R.opimus sargadensis 11 5/6 Semnan Damghan Hasan-Abad, Soltanieh Ghasem-Abad R.opimus sargadensis 7 2/5 Esfahan Badrood Badrood R.opimus sargadensis 17 5/12 Esfahan Habibabad Habibabad -- 0 0/0 Fars Neyriz Neyriz, Lar Total 50 19/31 5 9 17

north (Semnan P.) of Dashte-Kavir desert in center of Iran. TNF-毩, TNF-毬, IL-4, IL-10 and IFN-毭 might play role [20-22] Observations on the site of colonies showed that the great in severity and outcome of the diseaseL. major . There is also a gerbils dug their burrows near the agricultural lands e.g. correlation or specificity between strainsL. major and their sugar beet, cotton, corn, clover, barley, grape, pistachio, local natural animal hosts; hence, specific strain alfalfa, wheat farms and causes damage to crops. These can cause infection only in their proven regional hosts[23]. It territories showed small differences in burrow density but also correlated with the apparent specific resistance of local the number of gerbils was more stable in the south (Esfahan natural animalL. major reservoir hosts to cross-infection with foreign P.), in comparison to the north. Less density was found in strains of from other regions. This informationL. major Northern-Khorasan P., near the north-easternR. limitopimus of its should be determined by comparison studies on the distribution in Iran. There was no trace of in strains present in each great gerbil subspecies. Further Fars P. in southern limit of distribution of this species in genetic studies on these two subspecies could provide Iran. In northern slope of Alborz Mountain Chain (AMC), valuable information in epidemiology of the disease and can Golestan P., also less number of great gerbils was captured. be applied to human leishmaniasis and could lead to the In this area, the burrows of great gerbils were mostly located development of novel and specifically targeted therapeutic at foothills. This kind of habitat is a new record on the approaches. ecological habitats used by this animal. In this area, small This study demonstrated difference in density of the great part of populations was seen near wheat and barley farms. gerbil in different regions. This different density depends According R.to theopimus classification mentionedR. opimus in sodalis material andR. on many ecological features such as food abundance, opimusmethod, sargadensis two subspecies, and distribution and environmental factors and human R. opimusweresargadensis found in the great gerbil territories interventions. It shows that climate forcing synchronizes the studied. , with brown to yellowish dynamics of gerbils over large geographical areas[24]. They 2A/B (Figure ) was the dominant subspecies and found in suggested that based upon predictedR. opimus climate changes during all territories including Esfahan, Semnan, and Northern- the next century the densities of will be increased Khorasan Provinces around the Dashte-KavirR. desert,opimus and diseases such plague epizootics may become more sodalislocated in southern slope of AMC. Interestingly, frequent in central Asia. with hazel-nut color was found only in Golestan P. Sand flies which are vectors of the leishmaniasis use in northern slope of AMC. This province is connected to the burrows of great gerbil as the main resting and breeding northern neighboring country Turkmenistan, and seems to sites.Phlebotomus Great gerbil is the favorite host for blood-feeding be the southern limit of this subspecies in the region. L.of major sand flies. In all old World regions where Phlebotomus exists,R.opimus-P. it is transmitted papatasi-L. by sand majorflies of the genus 4. Discussion . association in Central Asia, Iran and Afghanistan are stable well- [25-28] R. opimus described zoonotic systems R.. Theopimus Leishmania In this study we found two R.subspecies opimus sodalisin two parasite can live in some tissues of for a long distinct geographical regions of Iran. R. opimus time, practically, thorough the lifetime of great gerbil sargadensiswas present in northern slope of AMC whereas without losing their virulence and availability for vectors. has occupied all territories in the southern slope Substantial part of the great gerbil population lives for more of AMC positioned around the central desert of the country. than 1 year, which is of great importance in epizootiology of AMC It seems the acts as a natural barrier between these two cutaneous’ leishmaniasis (the agent is preserved safely in the subspecies preventing gene flow between them. These two populations during the 7-8 month-long absence subspecies serve as the natural animal reservoir hosts of ZCL of active vectors). Great gerbil is a diurnal herbivorous in their territories[4,5]. animal, while most of the other species of this subfamily Presence of two subspecies in the country with noticeable are nocturnal seed-eating forms. As a diurnal animal the variations in morphology and ecology suggests potential great gerbil is guided by sight and has small thick conchae, genetic variation between them which might affect on ZCL while other gerbils are nocturnal species that are guided disease severity and outcome. Identification of genetic by hearing and have large thin conchae. That is important variation particularly the genes that determine various because sand flies usually pierce skin of the upper edge of outcomes such as developing asymptomatic,Leishmania symptomatic major (L. the conchae while feeding on gerbils, where short and sparse ormajor) severe disease upon infection with hairs make the skin more available for the short proboscis is highly recommended. Some researchers indicated of sand flies. Apparently thick ears of great gerbils contain that a number of genes of hosts (human or animals) including more blood and interstitial fluid, compared to the thin ears Mohammad Reza Abai et al./Asian Pacific Journal of Tropical Medicine (2010)800-803 803 of other gerbils, and sand flies prefer to feedR. on opimus the first. In J,et al. Spatial epidemiology in zoonotic parasitic diseases: insights general, most of the ecological features of favor gained at the 1st InternationalParasites Symposium Vectors on2 Geospatial Health in the existence of sand flies and the leishmanias in direct or Lijiang, China, 2007. 2009; :10. [8] Zhang YJ, Dai X. StudyZhonghua on the Liu situation Xing Bing of plague Xue Za in Zhi Junggar indirect way.L. Thatmajor is evidenced by complete coincidence of Basin of China study. 2008; the area of in the old world with the area of great 29 : 136-44.Walkers of the world gerbils, the latter being the only principal carrier of the [9] Nowak R. . 6th ed. Baltimore: Johns leishmanias. Hopkins University Press; 1999. Rhombomys opimus The relative effects of host subspecies and localityR. opimus on [10]Shar S, Lkhagvasuren D, Molur S. . IUCN red ectoparasite assemblages harbouredet by al the list of threatened species version 2, 2009. [Online]. Available populations were assessed by Tajedin [29]. They found from: www.iucnredlist.org. [Accessed on 29 September 2010]. Xenopsylla nuttalli [11]Molur S, Srinivasulu C, Srinivasulu B, Walker S, Nameer PO, only one fleaOrnithonussus species of bacoti and one mite Status of non-volant small mammals. Conservation species of across the area study. The AssessmentRavikumar andL. Management Plan (CAMP) workshop report . India: flea was found on both subspecies in all parts of the study R. opimus Coimbatore; 2005.The mammals of China areasodalis whereas the mite was found only on southern [12]Smith A, Xie Y. . Princeton, New Jersey: populations. This suggests that locality has particular Princeton University Press; 2008. ’ effect on the mite species but R. flea opimus subspecies. [13]Mallon DP. Rev The mammals15 of the Mongolian People s Republic. As a conclusion, because is the major principal 1985; : 71-102. carrier of the leishmanias and plays an important role in the [14]Rogovin K, Randall JA, Kolosova I , MoshkinRhombomys M. Social opimuscorrelates existence of the parasitic system of cutaneous leishmaniasis of stress in adult males of the great gerbil, Horm Behav ,43 in in the Old World such as Iran, additional field observations years of high and low population densities. 2003; : plus genetic structure studies are required for more detailed 132-9. Mammal species of the world [15]taxonomicWilson DE, and Reeder geographic DM. reference : A description of the great gerbil range, subspecies, rate L.of . 2nd ed. Washington and leishmaniasis infection, its collaboration with specific major London: Smithsonian InstitutionRodents Press;1993. and their identification keys strain over its large geographical range. [16]Etemad A. Iranian mammals: . Conflict of interest statement 1st ed. Publication of National Association of natural resources and human environment protection. Tehran: Tehran University Publication; 1978. We declare that we have no conflict of interest. [17]Goodwin GG.American Mammals Museum collected Novitates by the Legendre1082 1938 Iran expedition. Key to rodents.1940; Fauna: 1-17.of the USSR Acknowledgments [18]Vinogradov BS, Argiropulo AI. . Jerusalem: AIsrael field Program guide for for Scientific identifying Translations; of Iranian 1968, p.desert 230. [19]mammaliansZiaei H. We thank the staff of Health Centers in Esfahan, Fars, . Tehran: Iranian Environment Organization; 1996. Northern-Khorasan, Semnan, and Golestan Provinces, [20]Kamali-Sarvestani E, Rasouli M, Mortazavi H, Gharesi-Fard B. (TUMS) and staff of School of Public of Health for their Cytokine gene polymorphisms andCytokine susceptibility 35 to cutaneous contributions and assistance in collection of great gerbils leishmaniasis in Iranian patients. 2006; : 159-65. and their help in maintenance of field caught rodents at [21]Salhi A, Rodrigues VJr, Santoro F, Dessein H, Romano A, animal house respectively. This study was supported by Castellano LR, et al. Immunological and genetic evidence for a grants 87-03-27-7836 from Tehran University of Medical crucial role of IL-10 in cutaneousJ Immunol lesions in180 humans infected with Sciences to Mohammad A. Oshaghi. Leishmania braziliensis. 2008; : 6139-48. [22]Sakthianandeswaren A, FooteTrends SJ, Parasitol Handman E. 25The role of host genetics in leishmaniasis. 2009; : 383-91. References [23]Elfari M, Schnur LF, Strelkova MV, Eisenberger CL, Jacobson RL, Greenblatt LeishmaniaCL, et al. Genetic major and biological diversity among populations of . Microbes Infect from7 Central Asia, the Middle [1] Strelkova MV, Eliseev LN, Ponirovsky EN, Dergacheva TI, East and Africa 2005; : 93-103. AnnacharyevaRhombomys DK, opimus Erokhin PI, et al. Mixed leishmanialLeishmania infections [24]Kausrud KL, Viljugrein H, Frigessi A, Begon M, Leirs H, inmajor : a key to the persistence Annof Trop Med Dubyanskiy V, et al. Climatically driven synchrony of gerbil from one transmission season to the next. Proc Biol Sci Parasitol 95 populations274 allows large-scale plague outbreaks. 2001; : 811-9. 2007; : 1963-9. [2] Faulde M, Schrader J, Heyl G, Amirih M, Hoerauf A. Zoonotic [25]Oshaghi MA, Rasolian M, LeishmaniaShirzadi MR, tropica Mohtarami F, Doosti S. cutaneous leishmaniasis outbreak in Mazar-eInt Sharif,J Med Microbiolnorthern First report on isolation of from sandflies of a Afghanistan:298 an epidemiological evaluation. Expclassical Parasito urban cutaneous126 leishmaniasis focus in southern Iran. 2008; : 543-50. l 2010; (4): 445-50. [3] Akhavan AA, Mirhendi H, Khamesipour A, Alimohammadian [26]Oshaghi MA, Yaghobi-Ershadi MR, Abbassi M, LeishmaniaParvizi P, MH, Rassi Y, Bates P, et al. Leishmania species: Detection and majorAkhavan AA, Rahimi Foroshani A, et al. Detection of identificationExp by Parasitol nested PCR assay126 from skin samples of Iranian in J naturallyPub Health infected37 sand flies using semi nested-PCR. reservoirs. 2010a; (4): 552-6. 2008; : 59-64. [4] Rassi Y, Abai MR, Javadian E, Rafizadeh S, Imamian H, [27]Rassi Y, Oshaghi MA, Azani SM, Abaie MR, Rafizadeh S, Mohebali M, et al. Molecular data on vectors and reservoirBull Soc hosts Pathol of MohebaiLeishmania M, et al. majorMolecular Leishmaniadetection of turanica Leishmania infection Exotzoonotic cutaneous101 leishmaniasis in central Iran. due to andVector Borne Zoonotic Dis in the vectors 2008a; : 425-8. and reservoir host in Iran. 2010. [Epub [5] Rassi Y, Sofizadeh A, Abai MR, OshaghiLeishmania MA, Rafizadeh major S, ahead of print] Mohebail M, et al. Molecular detection of in the [28]Akhavan AA, Yaghoobi-Ershadi MR, Khamesipour A, vectors and reservoir hosts of cutaneous Iranian leishmaniasis J Arthropod-Borne in Kalaleh Dis MirhendiLeishmania H, Alimohammadian MH,Rhombomys Rassi Y, opimus et al. Dynamics District,2 Golestan Province, Iran. of infection rates in (Rodentia: 2008b; : 21-7. Gerbillinae) populationBull of anSoc endemic Pathol Exotfocus of zoonotic103 cutaneous [6] Mohebali M, Javadian E, Yaghoobi-Ershadi MR, Akhavan AA, leishmaniasis in Iran. 2010b; : 84-9. Hajjaran H, Abai MR. Characterization of leishmania infection in East [29]Tajedin L, Rassi Y, Oshaghi MA, TelmadarraiyRhombomys Z, Akhavan opimus AA, rodentsMediterr from Health endemic J areas10 of the Islamic Republic of Iran. Abai MR, et al. Study on ectoparasites of , the 2004; : 591-9. main reservoirIranian of zoonotic J Arthropod-Borne cutaneous LeishmaniasisDis 3 in endemic [7]  Zhou XN, Lv S, Yang GJ, Kristensen TK, Bergquist R, Utzinger foci in Iran. 2009; : 41-5.