Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 1 100,000 infec humans, that known well is It (1). lumen intestinal human the in manni, hart E. coli, E. polecki, E. moshkovskii, E. dispar, E. ( which of eight species, many contain intestinal Human prevalent. and non and pathogen including others but reports case as are homi Microspora as such protozoa these of some on Reports Iran. in man hu different 18 over for transmissions of Cases Introduction Key words:Key currently available on the molecular distribution of intestinal human amoeba Iran. in with infection study present the data, current the also and Iran in amoeba of thatepidemiology molecular of importance suggested is It country. especiallyspecies the in central northern and areas of Iran amoebiasi and molecularstudies,molecularBasedseemsstudythe isonoflow.it theseIranin that protozoa PCR as such methods some by intestinal human of distinctiontheappreciatedbetween prevalence high a show Iran. of parts Iran different in in amoeba performed amoeba intestinal including protozoa pathogenic microscopic Many Abstract Molecular Epidemiology of Human Intestinal in Iran in Amoebas Intestinal Human of Epidemiology Molecular Iranian J Publ Health, Vol. Vol. 4 JPublHealth, Iranian 2 i, aatdu coli Balantidium nis, E. histolytica E. . in wt a ana dah ae f over of rate death annual an with tions Iodamoeba butschlii and Endolimax nana Endolimax and butschlii Iodamoeba Dept. of MedicalDept. of Mycology & Parasitology, School Public Health,of etnl rtza ae en eotd in reported been have protozoa testinal - olwd () Te te hmn intes human other The (2). worldwide ht ass bu 5 mlin ae of cases million 50 about causes that common more are species pathogenic spp, Amoeba, 1 is the only pathological species in species pathological only the is . ylsoa cayetanensis Cyclospora - ae eieilgcl uvy o te rvlne f ua itsia ptoei ad non and pathogenic intestinal human of prevalence the on surveys epidemiological based Dept. of Parasitology, SchoolDept. of of KashanMedicine, of University Sciences,Medical *Corresponding Author: *Corresponding and 1 , No. , * H Hooshyar H * naeb moshkovskii Entameoba - 9 based methods, monoclo methods, based , Molecular , , Entamoeba Sep Such epidemiological studies on are confusing, mainly due to recently to due mainly confusing, are amoebiasis on studies epidemiological Such Entamoeba histolytica (Received 201 2 , , pp. , e pidemiology, Iran Sarcocyctis 10 ) reside ) 16 1 . moshkovskii E. - , P Rostamkhani P , 17 Mar Tel: +98 3615550021Fax:Tel: +98 3615551112 - - 201 - , E. E. dispar nal antibodies and the analysis of isoenzyme typing, however typing, isoenzyme of analysis the and antibodiesnal 2 ; accepted 12 accepted ; including intestinal amoeba, have been performed been have amoeba, intestinal including non and pathogenic tinal intes human of prevalence the on studies logical microscopic many present, to Up and prevention the con planning for important are amoeba, pathogenic especially protozoa, intestinal epidemio the parasitic on data of Molecular infection. epidemiology of concept and ledge intestinal human of proto epidemiology the to thod me this of Application tools. biological molecular techniques laboratory the in developments by driven been has infection zoa proto of epidemiology molecular the in Progress disease (3,4) humans in intestinal cause rarely non considered are amoeba tinal a b cmo aog rnas Cnieig the Considering Iranians. among common be may and trol programs. Tehran University University Tehran of Sciences, Medical Iran Tehran, o (.. mea rvltoie te know the revolutionised amoeba) (e.g. zoa E. moshkovskiiE. 1 , M Rezaian M , s due to rare histolytica iss due E. a infection the in Jul Available at: 201 2 ) . Differentialdiagnosis. can be done 2 http://ijph.tums.ac.ir , such as PCR and other and PCR as such , Review - Kashan, Iran E. disparE. pathogenic - - based epidemio based reviews the data the reviews ahgnc and pathogenic ispredominant Article protozoa, logy of logy . ------Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 hard copies of articles published in scientific jour scientific in published articles of copies hard out carried was the search manual Moreover checked. were articles selected of references search, of sensitivity the maximize To study. tothe words related databases key with transcription Persian and English the was Iranian in strategy searching The egy. "Iran", " keywords: "Mo following e.g. the and Amoeba" amoeba tinal of names the a including terms studies, related all identify combi to attempt an In published up to the 2011. 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Although sev Although 7). , Magiran ), al .: erhn were Searching , Intestinal "," Molecular Epidemiology of Human Intestinal Amoebas Intestinal Human Epidemiology of Molecular larstudies on utd to ducted and - cience based E. se ), ), ------ihd n 93 eaae ti seis rm the from species this harm separated 1993 in lished of basis re the formal On (10). biochemi typing isoenzyme the of analysis and antibodies monoclonal methods, PCR probe, DNA specific as such methods some by differentiated be could species two These species. identical morphologically but distinct cally histolytica histolytica Entamoeba Entamoeba Results scientific rese Iranian by congresses at presented topic this to related ticles ar scientific of abstracts the as well as theses, ate post and projects research of reports nals, E. histolytica/E. dispar histolytica/E. E. of isolates 15 of study comparative preliminary A for com 30% to >1% of rate bet separation no examination with stool microscopy routine the using on out carried been have studies epidemiological of number a Iran In amoebiasis. of epidemiology the in change satisfactory a plays as of diagnosis differential rate accu for tools molecular of use and Identification (13). male homosexuals the among common of prevalence local American, South and Central some Africa, South India, Mexico, Japan, (12). the worldwide than common more much and that cepted histol of separation the since changed has amoebiasis of epidemiology the mentioned, criteria the Given epidemio clinical in important is diag species two the of tion par the while species, pathogenic potentially a . itltc, . dispar E. histolytica, E. is non is oi ad ramn ad lo rm an from also and treatment and nosis plex was reported in different parts of Iran (5). Iran of parts different in reported was plex ytica less commensal less logical point of point view. logical a, im cal, and - - pathogenic ecito of description . dispar E. E. dispar E. Available at: uooia ad eei dt, a data, genetic and munological oee i sm rgos (e.g. regions some in However E. histolytica E. and E. histolytica/E.dispar E. E. dispar E. and Asian Pacific countries) Pacific Asian and sltd rm aaa, by Hamadan, from isolated s h peoiat species predominant the is one. species. Nowadays it is ac is it Nowadays species. en hm A prevalence A them. ween … . dispar E. and archers. an . histolytica E. http://ijph.tums.ac.ir Hence the differentia the Hence . itltc/. dispar histolytica/E. E. d Entamoeba Entamoeba dispar Entamoeba (11). . moshkovskii E. is especially more especially is r to geneti two are E. histolytica E. . histolytica E. a pub was p. such spp. complex - - gradu E. dis E. based dis E. is ------Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 n Hmde (ot o Ia) hwd ht 19 that showed Iran) of (south Hamidieh and of and Rezaian by Hoo made that observation Similar this in study area. previous be dispar were isolates of 95.45% that showed Tehran of dispar tica/E. isolates 21 on study Another (10). his were isolates of dispar of isolates 49 of diagnosis rential the diffe Karaj, Iran central of areas metropolitan major two and Tehran In (16). species two with infectio mixed a had area this in person of 3.7% were isolates of 7.4% region) subtropical and (tropical southern In isolates ofwere 5.9% region) (temperate northern In by bothspecie infected were uals as were isolate identified of 93% were isolate of zone) 3.5% temperate (semi area central In follow: as were areas different in passer cyst in species two inf of ratio The (16). Iran in ers’’ predo that demonstrated study This were 7.9% were isolates 92.1% sho Iran of gions dispar E. of diagnosis differential on Data re subtropical (15). regions other the gions) than and (tropical southern in higher of ratio that s showed Iran).This (southern Ahwaz of areas rural in seen was (8.3%) rate highest the Iran),while tral prevalence (cen Ardekan and Yazd Tehran, in minimum 0.6% was rate The respectively. Iran, of part southern and northern central, for 4.6% and with tion infec of prevalence the showed Iran southern and dispar histolytica/E. dispar all that showed technique PCR tolytica E. histolytica/E. dispar histolytica/E. E. Available at: Available . histolytic E. ha(06,sn PCR shyar(2006),using E. histolytica E. minant species found among the ‘‘cyst pass ‘‘cyst the among found species minant (14). A field study on the distribution of distribution the on study field A (14). and only one isolate (4.55%) was found to found was (4.55%) isolate one only and sn PCR using . One person (2%) had a mixed infection mixed a had (2%) person One . y oeua methods molecular by E histolytica/E. dispar histolytica/E. E . histolytica E. ope i feh to b PR in PCR by stool fresh in complex a 1) Ti fnig s iia with similar is finding This (17). E. dispar E. E. histolytica E. http://ijph.tums.ac.ir - FP eeld ht 6 (93.9%) 46 that revealed RFLP n 9.% f isolates of 94.1% and cyst passers in northern, central, northern, in passers cyst e ta a oa nme of number total a that wed E. . dispar E. . itltc/. dispar histolytica/E. E. d ispar r ie ifcin (16). infection mixed or , while only 2 (4.1%) 2 only while , n ua aes f Ahwaz of areas rural in - FP n 1 isolates 21 on RFLP n 88.9% and and 3.5% of individ of 3.5% and Iranian J Publ Health, Vol. 4 Vol. Health, J Publ Iranian n te remaining the and the isolates were isolates the a 07% 3.9% 0.78%, was s. . dispar E. E. histolytica E. ection rate with rate ection n ifrn re different in E. histolytica/E. E. . histolytica E. . histoly E. E. dispar. E. E. dispar E. s the is tudy was and E. E. E. E. n ------, . 1 , , No. h 8 smls containing samples 88 the of all that showed Iran, of part (Golestan) eastern north and Azerbaijan), (West northwestern tan), asympto of diagnosis the for PCR and kit detection antigen stool of study comparative A (19). all at there detected not was as them of six fied, histolytica/E. E. microscopical eight on (2009) al. et highi Hag by electrophoresis, PCR/gel by study A (18). the showed them of were none and isolated samples 16 that showed bad Gon of local areas rural and urban in A study prevalence infection(5). mixed showed (4.76%) ple for positive (4.76%) s hr te ae lcrpoei pten and patterns electrophoretic mobil same the share that strains amoeba of group a as defined is deme zymo A (HK). hexokinase and (GPI) isomerase phosphogluc (ME), enzyme malic were isoenzymes chosen The to the between used distinguish gel starch in electrophoresis Isoenzyme region. this in study more to need and Iran of parts other is finding were them .dispar histolytica/E E. 11 Bakhtisry), and (Chaharmahal City Chelgard in samples stool 655 From study this (2011).In al. et dispa that vealed re Iran in amoeba on study molecular only The 1:8 and 1:2.7 (16). Coast) (Ivory 1:46 and (Bangladesh) (Philippines), 1:8.5 ratios: the similar found have countries other in studies based the that than frequent more that much showed (2004) al. et Hooshyar to due amoebiasis and Iran in species predominant that showed studies these of All for negative cysts were am 9 E. histolytica E. , ples (90.48%) were positive for positive were (90.48%) ples Sep r iy nrh f rn uig C method PCR using Iran) of (north City ity for these enzymes (22). forenzymes these ity is a new study that published by Pestechian by published that study new a is 201 ai cs passers cyst matic E. histolytica/E. dispar histolytica/E. E. 2 nie h peiu rpre dt in data reported previous the unlike , pp. , E. histolytica E. E. histolytica E. mts (G) guoe phosphate glucose (PGM), omutase is a rare infection in Iran (Table1). Iran in infection rare a is 10 dispar - 17 Entamoeba E. histolytica E. E. dispar E. ape i Zhdn identi Zahedan in samples E. histolytica E. bevd B PR 0 of 10 PCR By observed. was more prevalent than prevalent more was n 1 and E. histolytica E. . histolytica E. rm etr (Luris western from . itltc/. dispar histolytica/E. E. whereas ratio is 1: is ratio E. dispar E. E. E. histolytica p ifcin in infection sp. and another sam another and E. dis E. (20). E. dispar E. and E. histolytica E. . dispar E. ad also and , 2) This (21). 12. Field 12. par E. dispar E. . dispar E. - positive pattern s the is 12 ; one ; E. is ------. Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 ysis anal Zymodem antigen PCR & detection Stool Methods basedPCR Diagnosis Method of et f DNA of ment to belong were rest XX) and dems:II,VI,VII,X,XI,XII,XIV th be pattern to zymodems proved Eight isolates. human from differ 23 of totally of the Twenty of (12). zymodemes20 than more of examination possi the was the characteris simultaneous By 13 tic banding pattern of PGM and GPI, it GPI, and PGM of pattern banding tic - ble to classify the amoeba in to one to in amoeba the classify to ble 2002 2004 2006 1999 1999 s o the of ose - Year ae tcnqe, isoenzyme techniques, based 2006 2001 2006 2011 2010 2008 2006 2003 2001 2001 E. dispar E. - - - - - 2005 2008 2007 2001 2001 Table 1: Table n zmdms described zymodemes ent Hooshyar . Tehran,Zahedan,Gon bayijan and Golestan Tehran and Tehran Kazerun Lurestan,West AzarLurestan,West Different regionsDifferent of Ahwaz &Hamidieh Prior to the develop the to Prior . histolytica E. Region studyRegion of Prevalence of Prevalence Northern regionNorthern Southern Southern region Tehran& Tehran& Karaj Central region Khorramabad et South South Iran al Hamadan Chelgerd Zahedan Gonbad Tehran Tehran Tehran .: (Iran) Iran bad Molecular Epidemiology of Human Intestinal Amoebas Intestinal Human Epidemiology of Molecular , while the while , - one out one ( zymo E. histolytica - - - No ofNo cases 101 21 49 15 23 88 10 16 53 21 16 8 2 8 8 showed that all of them were zymodem I (23, 24). I (23, zymodem were of them all that showed (AS2IR dispar E. of isolates dispar 8 of pattern zymodems on Study of diagnosis species. amoeba differential and infection amoebic consi (zymo and E. his E. Prevalence ratePrevalence (%) E. dispar E. dered the gold standard for the diagnosis of diagnosis the for standard gold the dered 90 10 from Hooshyar et al. (2001) study and two and study (2001) al. et Hooshyar from ee aayi o clue aobe was amoebae cultured of analysis deme) 4/55 95/45 3.54 91/37 - 7/4 88/93/7 26 74 4/8 90/54/7 and AS16IR) by Haghighi (2006) in Japan in (2006) Haghighi by AS16IR) and 3/5 4/1 93/92 4/9 92/13 5/9 94/1 ------E. dis E. slts rm ern n Kazeroon and Tehran from isolates 100 100 100 93/75 100 100 In Iran 100 100 9 Available at: 3 Mixed - 3/5 ------… http://ijph.tums.ac.ir Nazemalhoseini Nazemalhoseini etal. (18) Solaymani Hooshyar etal. (23,24) Rezaian and Hooshyar Nazemalhosseini etal. Kheirandish etal.(28) Pesthechian etal.(2 Pesthechian Hooshyar etal. (10) Hooshyar etal. (16) Hooshyar etal. (16) Hooshyar etal. (16) Hooshyar etal. (16) Hooshyar etal. (22) Haghighi A (29.24) Haghighi etal. (19) Sahebani etal. (25) Fallah M etal.Fallah (14) M Arthurs ( Arthurs Reference et al.(20) (27) - (5) Mohammadi No. et al. (17) ) 1 E. ) Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 ih gastrointestinal presented with age of years 50 and 31 between tients (34). Also Bangladesh from children young in ported preva (33 kovskii tica from it distinguish that characteristics all emetine), (e.g. drugs antiamoebic some to resistant rant waste and (12). worldwide water clean sewage, in mon free a primarily considered is organism This available. diagno thropho from moshkovskii moshkovskii Entamoeba Entamoeba metronidazole with fully patie the the Finally months. to prior months 4 presen and States, United the Sistan the in P born was who tient Solaymani by cerebros tica provide can of case and first The diagnosis. rapid amoebiasis intestinal extra of increa are PCR methods Molecular (4). autopsy or biopsy of time the at organism the of detection or thods me serological by made often most is and portant im is amoebiasis intestinal extra the of Diagnosis of zymodem predominant the dispar E. of I zymodeme that suggested we results these on dispar E. histolytica E. as (74%) samples 17 and diagno were (26%) ples (2002 al. et Sahebani by medium Robinson's in cultured Iran, of south of isolate 23 of analysis Isoenzyme rov , oee a eet td rpre a high a reported study recent a however ), Available at: Available and - , can be cultured at room temperature, and is and temperature, room at cultured be can , living amoeba and has been found to be com tobe has found and been amoeba living ince of the southeastern Iran, immigrated to immigrated Iran, southeastern the of ince nehlts igoe b PR f the of PCR by diagnosed encephalitis lence (21.1%) of (21.1%) lence . moshkovskii E. . dispar E. ain f ies hd iie Ia fr 3 for Iran visited had disease of tation sis nowadays some PCR based PCR some nowadays sis a rrl be son o net humans infect to shown been rarely has E. dispar E. ot frs Fr aig differential making For forms. zoite pinal fluid of an Iranian patient was done was patient Iranian an of fluid pinal were I, XVI, XVII and XVIII (25). Based (25). XVIII and XVII XVI, I, were iia t ohr subt other to similar were II and XIV and, zymodemes of zymodemes and, XIV and II were - oamd e a. ( al. et mohammadi singly useful tools for the diagnosis the for tools useful singly (30,31,32). Up to present to Up (30,31,32). and http://ijph.tums.ac.ir a rcnl dtce i pa in detected recently was - ypos n utai (35). Australia in symptoms . histolytica E. sam six that showed 2005) lo s o distinguishable not is also E. moshkovskii E. E. moshkovskii E. sed as sed (26). E. dispar E. t a tetd success treated was nt E. disar E. Iranian J Publ Health, Vol. 4 Vol. Health, J Publ Iranian oia cutis is countries ropical Entamoeba histolytica Entamoeba Entamoeba histoly Entamoeba . Zymodemes of Zymodemes . 20 n t cs or cyst its in - contaminated 08 Entamoeba - has been re been has is osmotole is methods are methods in Iran.in Baluchestan . h pa The ). E. histolyE. E. mosh E. such as such in ------1 , , No. histolytica/E. dispar/E. moshkovskii with dispar/E. histolytica/E. infected person 16 of Out that showed Iran in study one latest A sequencing. by firmed of presence study this In of infection mixed were moshkovs 3.5%) 2( and for positive reported 53(91.4%) 2(3.5%), assay single using examined samples stool re A (20). person healthy a in recorded was 1.1% of with infection of prevalence low a moshkovskii studied other and of parts some in least at that, showed These 1.9%(37). India 24.7% (35), Australia (36), 13% world Africa the South as over such all studies many from reported spe this of isolates human date, To ing light microscopy. light ing the on based were detec done previously surveys all and Iran in amoeba intestinal nonpathogenic the of epidemiology molecular the on data reliable no tica mistak avoid to harm taken be to considered are spp. amoebas these Although 9). 1, (5, Iran around re was >70% to 1% between rate prevalence and A Iran. in human of butchlii Iodamoeba Endoli hartmani, Entamoeba amoeba coli, (e.g. species amoeba intestinal Various intestinal Other of detection culturemethods. by detection the for moshkhovs PCR using Despite of the Iran. in amoeba epidemiology molecular in point important an is This Iranians. with infection that suggest we study of this Considering were moshkovs that samples sixteen micro from showed PCR single Khorramabad, in examination scopic et td i Ia re Iran in study cent 9 te astv aet o amoebia of agents causative the , , Sep tion of cysts or trophozo or cysts of tion E. moshkovskii E. cpcly po scopically max nana max 201 kii kii kii 2 , respectively. One person (1.73%) had (1.73%) person One respectively. , ad 5 9.5) were (93.75%) 15 and , , pp. , may be a true human parasite. In Iran, In parasite. human true a be may n rn n atmt a dn fr its for done was attempt no Iran, in ) are frequently found in the stool the in found frequently are ) 10 - 17 otd n ifrn population different in ported E. disparE. iie 1 62% was (6.25%) 1 sitive, a b cmo aog the among common be may E. histolytica, E. dispar E. histolytica, E. n igoi with diagnosis en otd ht u o 3,825 of out that ported E. moshkovskii E. and es cr sol be should care less, ites in stools by us by stools in ites E. moshkovskiiE. yt b micro by cysts E. moshkovskii E. is a been has cies . dispar E. - i. hr is There sis. the world, the round Entamoeba E. histoly E. was con was - and round 14 PCR (28). (27). E. E. E. E. E. - - - - Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 in. hs a b cniee a a important an as considered be may This nians. on Ira among common be may species this study with tion the of results moshkovs the to According of tion rec in method important very a have typing that enzymes zymo cultures(40).However xenic bacterial in present with interaction are to zymodemes due remaining The origin. amoebal zymo in I zymodem for XIX and XIV II, modem only that (Zy typing for isolate valuable are four zymodems suggested studies subsequent Iran, in of demes Zymo of demes that revealed studies previous Although and of prevalence true the determine to done be should future study the epidemiological molecular Thus, ameobiasis. of the burden predict clinical to important very is community the of prevalence true the of Identification speculation. of matter a became has preva true and nite r a is to due amoebiasis and Iran in species predominant the is dispar E. that seems It study, non with infected actually were stool their in detected cysts with individuals asymptomatic of majority A the dispar E. histolytica, between distinction appreciated recently the to due mainly confusing, is amoebiasis of ogy wo the of parts by examination stool micro on done previously veys re the this to almost due of is condition explanation probable the most The species. on data the since comprehensive no epidemiol are Iran. in There concern health public a still (e.g. are species) infections protozoan Intestinal Discussion 15 - E. dispar E. pathogenic are infection in Iran (16, 18, 20), but the defi the but 20), 18, (16, Iran in infection are demes appear to be of bacterial rather than rather bacterial of be to appear demes scopy, not only in Iran, but also in the other the in also but Iran, in only not scopy, E. dispar E. kii g of ogy E. histolytica E. eaain of separation E. dispar E. n rn e ugs ta hmn infec human that suggest we Iran in , especially in the tropical area of Iran. area in tropical the especially , rld, because the current epidemiol current the because rld, as a separatespecies. as a . dispar E. . dis E. and . histolytica E. were I, XVI, XVII and XVIII and XVII XVI, I, were ec rt of rate lence were II and XIV and, zymo and, XIV and II were E. E. moshkovskii Hooshyar par E. histoly E. E. histolytica E. Bsd n molecular on Based , (23,9.h others )(12,38,39).The E. histolytica E. et n rn especially Iran in tica al liance of all sur all of liance E. histolytica E. .: . Molecular Epidemiology of Human Intestinal Amoebas Intestinal Human Epidemiology of Molecular and . histolytica E. infection in infection E. histolytica E. Entamoeba and only and E. dispar E. dems ogni still E. E. ------Ethical considerationsEthical Iran is necessary. identifica molecular the also and of the as waterborne considered are amoebas these so human, impor intestinal an human of knowledge proto our on Based molecular diag So, controversial. diagnos and difficult be differential that as established well diagno been has It Iran. e molecular the in point References of conflict no interests. is there that declare authors also The and review onthis manuscript. suggestion this for apprecia material the pro vided who authors all to grateful are authors The Acknowledgements authors. the by served ob completely been have etc) redundancy, sion, falsifi data Informed misconduct, plagiarism, Consent, (Including issues Ethical E. hart E. Entamoeba 3 2 1 nosis of infection with these protozoa. these with infection nosis of . . . ain dul pbiain n/r submis and/or publication double cation, zoa epidemiology, water and waste water has water waste and water epidemiology, zoa i o sm hrls hmn mea such amoeba human harmless some of sis i tos a b ueu fr t for useful be can tools tic lr C, imn L (91. Ribosomal (1991). LS Diamond CG, Clark health Public (2011). M Norhayati SA, Tengku Te Amoebas. (2011). M RezaianH, Hooshyar in o r rjad fr i helpful his for A Ardjmand Dr to tion at oe n rnmsin f mea to amoeba of transmission in role tant manni Bi genic’ ‘nonpatho and ‘pathogenic’ of genes RNA 222. in amoebiasis Ma of importance clinical and Sciences Medical of Publica University hran parasites. Investigating the occurrence the Investigating parasites. och Parasitoloch laysia: A review. 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