Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 Tehran University of Med of University Tehran Keywords: stage. invasive the during occurred phil The . in reported been never had which ment Conclusion: dose eosinop blood the prisingly, 8 the larvae between eosinophils blood in difference significant a was There detected. was encystment va average in found were The worms The 78.03%. (76.47%). was months) (5 muscles recovery worm in found mostly were they PI, 15 day At fat. dorsal and heart lung, 24 Results: blood with eosinophilia. along of AL3, migration the order tostudy Methods: stage larvae ( gra i migration regarding Information . ABSTRACT Original Article 73 Background: Sciences Public Sciences Larva Migration and Eosinophilia in MiceExperimentally in Eosinophilia and Migration Larva tion by recovery of worms from the whole body of mouse after oral infection with infection oral after mouse of body whole the wormsfrom of recovery by tion hr PI; they were recovered from various organs including liver, mesentery, esophagus, diaphragm, esophagus, mesentery, liver, including organs various from recovered were they PI; hr http:// tums.ac.ir http:// - W. Saksirisampant W. - group (25 larvae). (25 group (average 9.33% (average At 1 hr post infection (PI), the larvae remained in the stomach, thereafter at 3, 5, 7, 10 and 10 7, 5, 3, at thereafter stomach, the in remained larvae the (PI), infection post hr 1 At Mice wer Mice AL3). The pe The AL3). 1 ie ws novd by involved was Liver . 3 2 Gnathostoma . . a tion Dept. of Microbiology,Dept. of of S Faculty Medicine, In Dept. of Pathology,Dept. of Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand Parasitology, Dept. of Faculty of Medicine, University,Chulalongkorn Bangkok, Thailand e orally infected with AL3 and histological study of organs was invest was organs of study histological and AL3 with infected orally e fected With fected i cal + spinigerum 6.25%) and the 15 larvae 15 the and 6.25%) r centage of blood eosinophils was examined in parallel. in of eosinophils blood examined was centage 1,2 hils (average 19.00% (average hils * Iranian J Parasitol: Vol. Parasitol: J Iranian Corresponding author:Corresponding , N. Choomchuay N. , (Received , Larvae, Worm, Larva migration, Eosinophil migration, Larva Worm, Larvae, , G. spinigerum G. causes larva migran in human which is endemic in Southeast in endemic is which human in migran larva causes Iranian J Parasitol J Iranian 17 Gnathostoma spinigerum Gnathostoma Open access Journal at Journal access Open http:// ijpa.tums.ac.ir http:// Mar Thanomsub s limited. In this study, we investigated the parasite m parasite the investigated we study, this In limited. s 201 2 ; accepted + - 3 hogot h suy W dtce lra encyst larva detected We study. the throughout 7 infected groups (average 22.66% (average groups infected , K Kraivichian K , , No. , Email: rinakharinwirot University, Bangkok, Thailand 2.92%) were not higher in the higher infective higher the in higher not were 2.92%) 3 1 , 20 , 21 [email protected] 1 Jun 2 , pp , 201 Available at: . 73 2 the liver at every time point. La point. time every at liver the ) - 3 81 , *B. Wongsatayanon *B. , highest percentage of eosino of percentage highest Iranian Society of Pa of Society Iranian http://ijpa.tums.ac.ir + http:// isp.tums.ac.ir http:// advanced third 11.03%). Su 11.03%). i gated in gated rasito r- r- i- - - l ogy Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 (1). R hispidum t of otherapy as usedeffectiveness chestudyingthemodelsoffor been have Rodents limited. termspathof regarding mation treat of effectiveness the of d the forindicator early philiais animportant hallmark which canbe an (17 infection past and sent differentialrologicalnot canbetests S test.serological positive a eosinophiliaand wa fresh cooked undeand/orraw ofhistory onbased is nosis skin lesion a orbiopsy specimen. presumptiveA fromdiag recovery worm on based is tion infec of diagnosis definite The (16). in observed been has organs visceral volved mo to mild and ocytes, infiltra eosinophilicand/or fatal near cause also othersandsymptoms (9 su pruritus migra i.e. migrans, doc visit usually infection this with Patients (1,7). (AL3) larvae fresh water contain by co (3 America and Europe co also now accidental an hu andetc.) mink ferret,weasel,boar, ( of tode nema a actually is worm The (2,4). Thailand is common portedhumanin infection (2,3). of species Seven Amer South in countries certain and G Introduction Available at: o t n clr novmn, og, hematuria cough, involvement, ocular in lt dents are accidental hosts similar to humansaccidentalsimilararehostsdentsto n tion of eosinophils,fibroblastshistiof andtion sumptionof insuffi (7,8). In addition, visceral larva can r can larvavisceraladdition, In (7,8). menin and onre o Suhat sa I has reportedinalsobeen It Asia. Southeast of manycountries in endemic is nathostomiasis n . spinigerum G. o G. goencephalitis(6,14,15).heavy A iee a eegn dsae in disease emerging an sidered genesis,im ot 14. ua infec Human (1,4). host i seis n toe of those and species his http://ijpa.tums.ac.ir os ih subcut with tors e fs cnupin blood consumption, fish ter nipponicum o, a, ie, in leo lion, tiger, , , oy wlig ih an and pain with swelling tory Gnathostoma ing advanced third stage - i - - para Saksirisampant agno 6). Infec 6). 13). The diseaseThe13).could eae dm o i of edema derate muneresponsestillis ciently cook - which 19). Blood eosin Blood 19). ment ie neato in interaction site infec sis and sis parts of Me ofparts ae en re been have Globally tion is caused is tion tions 82) Info (8,20). a s on in found is eu larva neous tween prtween evaluation ed orraw mans are mans i et al et a (1,2). ca (21 in is tion , pard, most .: - x 26). ico Larva Migration and Eosinophilia and Larva Migration m- G. o- n- e- e- e- r- r------l, egig appro weighing old, G. usedWeadult maleSwissalb d previously as with pepsin artificial mice ( eelsinfected of (AL3)obtainedwerelivernatuthe of from of larvae stage third infection Advanced spinigerum Experimental Materials and Methods in parallel. examined eosinophilswas blood percentageof the from The wormsAL3. withinfection oral after body whole of recovery by parasite studymigration to model mouse the used We nml o te aut o Medi of Faculty the Chulalongkorn Un of animals was study approved Thistheby Et 0. Day at infection the to or stainsolution (28).Theyalsowere checked pr Field’s with stained and tube microhematocrit were taken from the ophthalmic vein by using a pe the ce as larvae 25 and 15 8, eosino with infected Blood mice of groupsthree inexamined eosin. were counts and hematoxylin ogyfor processing, sectioning and stain patholformaldehydetosentfixedinwereand a under then examined plates, glass thick two between pressed r separately were and muscles organs The migration. study to order in eosi sacrificed were they intervals time different At blood and la viable 8 with infectedindividually thirty from recoveredhistology were Worms o recovery, Worm hygienic with a de and pellet under an the reared environment, were They gastricintuba by AL3 25 or 15 8, with fed phil counts phil ntage of total white blood cells. Blood smears - held for 3 hr prior to infection. i mal house and fed ad libitum with food libitumwith ad fed and house mal - Monopterus al hoiae tp ae. od was Food water. tap chlorinated n oyabnt shoe polycarbonate in i versity, Bangkok Thail h tro irsoe Tissues microscope. stereo x icsc … mtl 3 g. hy were They g). 34 imately ommitteedealing with ba e oe, lcd and sliced moved, ) by dige by ) i no micenoweeks(8 e cie (27). scribed . spinigerum G. - th r s - vae each. vae tion with tion oe in boxes ree mice ree ingwith and. cine, tion. rally phil 74 n- r- i- - Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 1 Mo 15 d 7 d 5 d 3 d 24 hr 10 hr 7 hr 5 hr 3 hr 1 hr tion Post Time Total 5 Mo 3 Mo sal fat, Mus; fat, sal Uro;Muscle, Urogenital Sub; organ, Subcutaneous tissue, Br; Brain Abbreviations: i n Larva migration Larva Result to be significant different. dent Stu by analysis determined was significanceStatistical Statistical Table 1: Table 75 fe c- - studied Mice No. t test. A value of value A test. s 33 1 1 2 2 2 4 5 4 3 3 2 2 2 Mean percentage of larvae recovery in various locations at different time intervals after infe after intervals time different variousin at recovery locations of larvae percentage Mean St; Stomach, Eo; Esophagus, Li; Liver, Mes; Mesentery, L Liver, Stomach,St; Li; Mes; Eo; Esophagus, Recov worm Total 15(93.75) 16 (100) (78.03) (93.75) (92.50) (81.25) (70.83) (45.83) (62.50) (56.25) (62 (87.50) (87.50) (68.75) 206 11 10 10 11 30 37 26 17 9 7 7 .50) ered P <0.05 was co was <0.05 (40.00) (85.71) (57.14) (80.00) (100) (9.09) 16 12 St 4 6 4 1 ------Iranian J Parasitol: Vol. Parasitol: J Iranian (10.00) (18.18) Eos 1 2 ------n sidered (15.39) (63.64) (70.00) (55.56) (30.00) (14.29) (27.27) (3.33) (8.11) (5.88) (6.67 Li 1 3 4 1 7 7 5 3 1 3 1 - - - tio ) n with 8 larvae 8 n with - (14.28) (13.33) No. wormNo. Mes 1 2 ------7 , No. , (10.00) (28.57) (18.18) ragm (Table 1). cludingesophagus,diaphheartandlung,liver, in organs various to moved they Thereafter, but (13.33%) mos mesenteries and (6.67%) liver migratbeganwormsthe PI, hr 3 (PI).At were stillfound theinstomach hrpostat 1infec larvae All migration. study to used was each larvae 8 withinfected mice of group The Lu 1 2 2 ------3 recovered (%) andrecovered (%) , 20 , wr sil n h soah (80.00%). stomach the in still were t (9.09) 1 He 2 1 ------, pp , u; Lung, Diaphragm, Di; Do He; Heart, Dfat; Available at: . 73 (10.00) (18.18) Di - 1 2 ------81 (33.33) (10.00) Dfat (5.88) 1 3 1 ------Location http://ijpa.tums.ac.ir (86.67) (83.78) (80.76) (76.47) (36.36) (20.00) (11.11) Mus 26 31 21 13 4 2 1 ------(5.88) Uro 1 ------(3.33) (2.70) (5.88) ing to ing Sub 1 1 1 ------tion c- - - (6.67) (5.40) (3.85) Br 2 2 1 ------r- Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 small lymph small inflamm and chronic larvae tissue fibrous thick the by surrounded that were showed thereafter sections in shown as PI month one and 20 Day during observed was cyst one larvae a gross The shown). not (Data PI 11 Day in observed first was organs invading larva around infiltration cell inflammatory The organs ed recov of larva Encysted C) p anterior of up the B) Close larva Whole A) recovered larva(AL3) third stage vanced 1: Fig. bulb ( head onthe hooklets of rows four with larva stage third were they o Mo sacrifice. phology to prior abnormal observed no was death However, brain. the in found be could larva the PI, months 5 and 3 ani 3 only involve subcutaneous demonstrated mice, infected 33 the all Of s invaded worms the PI, 15 Day At experiment. the throughout infection after point time every at observed be could and PI hr 3 at liver the in appear to ed star larvae Noteworthy, (20.00%). abdominal diaph in recovered liv the in observed still were larvae the of 70.00% PI, 5 Day At organs. other to migrated all almost Then stomach. the in still were worms few a PI, hr 24 and hr 3 During Available at: ered during 1 month PI from the inva the from PI month 1 during ered encystments with one larva coiled inside coiled larva one with encystments mg of Image mi f o cytes seen in various organs for organs various in seen cytes http://ijpa.tums.ac.ir rtd om idctd that indicated worms grated buaeu tsu (5.88%). tissue ubcutaneous a oy el, predominantl cells, tory am 1.0) n i the in and (10.00%) ragm ntotm spinigerum Gnathostoma Saksirisampant ntotm spinig Gnathostoma Fig. Fig art/head bulb art/head C Histological 1C. .1A, 1B) (4). .1A, p pearance of pearance er and first and er ment. At 1, At ment. et al et t or ity .: mals e rum a Larva Migration and Eosinophilia and Larva Migration d- d- r- t- y others (data notshown). others (data ( liver diaphragm, example, P infec vae (9.33% group fected diffe significant was There AL3. 15 with infected those from 47% was study this in percentage highest The eos blood 0% from ranged (aver infection the to prior mice in eosinophils blood of percentage The eosinophilia Blood larva surrounding the ous tissue fibr dense showing PI, months 5 at Liver B) ma infla showing PI, 20 Day at Diaphragm A) spinigerum thostoma 2: Fig. - value = 0.001)(Fig.3). = value tory cell tory age 1.50%, age Histological sections of encysted of sections Histological t ed group (average 22.66% (average group ed inophils reached a peak at Day 13 PI. 13 Day at peak a reached inophils s infiltration r ence between the 8 larvae in larvae 8 the between ence + larva in larva in organs: 1.38%). After i After 1.38%). … + 6.25%) and the 15 lar 15 the and 6.25%) Fig. A 2) and 2B) 2A, n fection, the fection, + 11 .03%, Gna - 4% m- 76 - - - - Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 8,31). We therefore used 8 larvae as a feeding a as larvae 8 used therefore We 8,31). 1 only was humans in found r common (ranged 40% 13.33% only from was recovery worm age ave the that reported studies These therapy. chem of effectiveness the of exper studies mental in used were rabbit) and de mouse (rat, past the Over adulthood. to mature to unable were They 1). (Fig. stage larvae third advanced were recovered worms of host definitive a not is mouse the that confirmed study Our migration Larva Discussion la 15 25 the and between signif statistically no was there ( animals infected di icant seen not 19.00% (average were eosinophils blood pe of higher centage a studied, was group larvae) (25 infe higher the when Interestingly, 77 Fig. 3: f ference between the 8 and 25 larvae 25 and 8 the between ference Percentage of blood eosinophil at different time intervals of mice i of mice intervals time different at of eosinophil blood Percentage e otd idn o wo of finding ported - 40% (32,0. h most The (23,29,30). 74.00%) + P - 2.92%). There was a signi a was There 2.92%). . spinigerum G. value = 0.001). = value r vae - Vol. Parasitol: J Iranian infected groups. infected - i c cant difference cant wrs (1,6 worms 3 ds rodents ades, ic al the all since m numbers rm c tivity dose tivity However, o- r- r- f- i- - - larvae 7 , No. , could be found in the liver at every time point time every at liver the in found be could (24 destination first as liver the reach to larvae the for necessary not was f move other nipponicum with infected Gnathostoma animals studied who investig other with concurred results Our liver, esophagus, from diaphragm. and worm heart lung, PI, recovered hr be 5 could At viscera. many through migrate freely and wall stomach results the penetrate Our 1). (Table du that showed infection oral after day first the during hours different at covery a were sacrif first the er, (21 sites other to migrating tually 7 of period a for remain would worms where organ first the was liver the that suggested they and liver to stomach spinigerum G. Prev gration. m larval of study for recovered 2 be could that least at of estimate an with dose 3 , 20 , 1 fter 1 fter reely from stomach to many viscera. It viscera. many to stomach from reely 2 , pp , Atr neto, h lra could larvae the infection, After . Available at: . - 73 i pce, i.e. species, r worm studied We PI. week 2 ous investigators showed that all that showed investigators ous - larvae 81 r ing 3 ing i il ae i toe studies those in dates cial n - fected with 8, 15 15 25 8, and with fected 5 hr PI, the worm could worm the PI, hr 5 ut is mv from move first must - http://ijpa.tums.ac.ir eve before days 14 . hispidum G. - 6. u larvae Our 26). - 23). Howe 23). - worms 3 and a tors G. n- v- e- i- s Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 doses to 8, 15 and 25 larvae for study of blood of study for larvae 25 and 15 8, to doses parenchym liver di severe showed Autopsy shown). 2 day on died five infection, la 30 of level high with mice seven the Of eosinophilia Blood host d ordertoevade in (su superficially more move m keep humans in larvae the that observed been never has humans in human encystment worm with individuals) G known. well is gnathostomiasis, infected in 50% subc of frequencies inve high the contrast, In other 24,29,30). by rats and rabbits Larval infected the in 2). observed also was encystment (Fig. i infiltration chronic cell moderate flammatory to mild with fibrosis others. and liver tissue, subcutaneous diaphragm, as such sues ti multiple in detected were larvae Encysted mostly (76.47% larval of site involvement major the were muscles ment, (5.88 PI on 15 tissues Day subcutaneous in larvae found the first addition, were In 33. of total a among mice 3 only in migration larvae subcutaneous (21 rabbits and rats mice, of models animal in others of findings an mice our in volvements i eye detect not could We (6,7,10,12,13,15). others and eye system, nervous central tract, studies urogenital lung, liver, Previous including organs visceral var to (32). migrate can parasites that factors showed other mechan defense and host infection, of of dose and a duration is by regulation influenced hosts process complex infected The in population helminth observations. vious 45.58% (ranged 78.03% was mice our in ave found worm The recovered. worms’ the of numbers different with study the throughout Available at: utaneous migratory swelling (approximate swelling migratory utaneous found by Day 15 and months 15 5 PI. Day found by - 100%) which was higher than in pr in than higher was which 100%) ) (,,,13,4. e hypoth We (1,6,8,31,33,34). . http://ijpa.tums.ac.ir . hrfr, e eue the reduced we Therefore, a. They were enclosed by dense by enclosed were They ) Truhu te exper the Throughout %). - 3 2, 0. e found We 30). 29, 23, - e was which 86.67%) Saksirisampant fenses (31). ti are with agrees this d b cutaneous tissue) cutaneous - P (aa not (data PI 8 s iaos (22 tigators i nathostoma grating and grating s ruption of ruption ae total rage et al et e sized .: r i ous ism vae Larva Migration and Eosinophilia and Larva Migration n- n- e- s- i- - (average 22.66% la (average 15 the in than higher 19.00% e (average sinophilia blood study, our in used was larvae) dose (25 infective higher the when Surprisingly, eosin blood maximum 6% only Rats i stimulation. eosinophil for protein site pa of amount the affect loads infected larval ferent larva 15 Di group. infected larva 8 the the in than group in higher was percentage count average the However, group. ed infec AL3 15 the in detected was which 47% hig The PI. 13 day on was seen animals infected our in eosinophils o blood in then peak The was served. weeks subsequent the during eosinophilia blood of Fluctuation eosinophilia. ht bod cells cases in only patients in decrease eosinophils blood white which 5% than p 70% about gnathostomiasis, human In lation. stim eosinophilic for allergen amount low a e the to due burden parasite low indicate not does count eosinophil low a that suggest findings Our PI. months2 within levels low to d gradually it plateau, a activation. eosinophil eosin blood the after However, governing events ry regulat the of understanding advance to and kno our expand to help could infected animals, dose parasite high with eosinophils which by spinigerum mechanisms of Elucidation (37,38). fections and chronic in o which cells immune of depression in i CD4 of subset interleukin on pendent d is infection parasitic by induced sinophilia e to differentiation and tion bone cells stem the pluripotential marrow or cells responsiveness progenitor of hematopoietic point maximum a to n n a fected with 3 larvae of larvae 3 with fected ovmn o te Th the of volvement tients demonstrate tients et perce hest n nue lw rdcin of production low induces cystment of worms, thus causing thus worms, of cystment n /or heavy dose of parasitic i parasitic of dose heavy /or ae n lo esnpis was eosinophils blood in tage + may exceed 50% of circulating of 50% exceed may helper T cells T helper + 11.03%). This may be due be may This 11.03%). … blood eosinophils blood - - e rsos cn result can response 2 poue by produced 5 rae ad dropped and creased G. nipponicum G. (1,8,20,31). r a ifce group infected vae osinophil + Ti mgt be might This . 2.9 o (36). Excessive (36). philia reached philia 2%) was not was 2%) o o prolifer to hla (25). philia (35). showed c w curred higher B ledge Th lood E r 78 G. of o- n- o- o- b- u- - e- a- a- f- t- 2 Downloaded from http://journals.tums.ac.ir/ on Tuesday, October 09, 2012 References interest. t that declare authors The T Mr. and Sarachana Tewarit Mr. indebt are We su technique histological and suggestion Srinakh Medicine, of Faculty Anatomy, of Pansir Department nsuwan, Professor thank We Thailand. 10330, Bangkok Thailand, of Council Research N by supported partly was study This Acknowledgements for model animal orstudies. oftreatment other drug evaluation poorer them made have th fact the be not fu for topic be should encystment stimulate that Mechanisms encysted. parasites the when level du occur could eosinophil blood high a addition, infective the on depending plateau a reaches eosinophilia blood the in level dose The high worm in seen experience is philia not eosin blood high study, our do In encystment. humans trast, co In occurred. encystment larva and hosts accid are mice that confirmed We study. infe post hr 3 at tected d was involvement Liver viscera. various to infection conclusion, In decl and stage du proportion greater in pr sent be might eosinophilia stimulated which of removal worms surgical or treatment drug after 79 a r r ei Srbosti o te nml handle. animal the for Siriboonsitti wekit ther study and have practical value. It may It value. practical have and study ther n te naie tg ad rp o low a to drop and stage invasive the ing 1 81,1. h ptnil components potential The (8,10,31). . Gnathostoma h Fcly f h toia Medicine, tropical the of Faculty the Sucharit S. editor. The In:25 review. gnathostomiasis.human A DaengsvangS. am Press; 19 Mahidol University. Bangkok: Krung s ine in the e the in ine our finding indicated that after that indicated finding our a t ave nytd n mice in encysted larvae at rinwirot University for her for University rinwirot 86. av cn m can larva c Gnathostomaspinig n P here is no conflict of conflict no is here in hogot the throughout tion . 124 cysted stage cysted - Iranian J Parasitol: Vol. Parasitol: J Iranian netd animals. infected r - ing the invasive the ing 47. th Anniversary of i rt freely grate oe In dose. (39). e rum Pha i a p tional port. ental and o- n- e- e- i- - 7 , No. , 3 , 20 , 1 5 4 3 2 11 10 9 8 7 6 2 ...... , pp , . . n importeding disease. ChiodiniPL. 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