Research Article The Effect of Anthelmintic Treatment on...

Research ArticleThe Effect of Anthelmintic Treatment onCoccidia Oocyst Shedding in a Wild Mammal Host withIntermittent Cestode Infection

Radovan Vaacuteclav and Jana BlaDekovaacute

Institute of Zoology Slovak Academy of Sciences Dubravska Cesta 9 845 06 Bratislava Slovakia

Correspondence should be addressed to Radovan Vaclav radovanvaclavsavbask

Received 25 July 2014 Revised 4 November 2014 Accepted 5 November 2014 Published 20 November 2014

Academic Editor Gadi Borkow

Copyright copy 2014 R Vaclav and J Blazekova This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

While hosts are routinely exploited by a community of parasite species the principles governing host responses towards parasitesare unclear Identifying the health outcomes of coinfections involving helminth macroparasites and microparasites is one area ofimportance for public and domestic animal health For instance it is controversial how deworming programmes affect incidenceand severity of such important microparasite diseases as malaria One problem is that most study systems involve domestic andlaboratory animals with conditions hardly comparable to those of free-living animals Here we study the effect of anthelmintictreatment on coccidia infection intensity in wild Alpine marmots M marmota Our results lend support to the hypothesis thathelminth infection has a positive effect on concurrent microparasite infection However our work also points to the fact thatwithin-host interactions between helminths andmicroparasites are context-dependent and can turn to negative ones once helminthburdens increase Our study suggests that coccidia benefit from intermittent helminth infection in marmots due to the protectiveeffects of helminth infection only during the early phase of the hostrsquos active season Also the marmotrsquos response towards coccidiainfection appears optimal only under no helminth infection when the host immune response towards coccidia would not becompromised thereby pointing to the importance of regular intestinal helminth elimination by marmots just before hibernation

1 Introduction

Under natural conditions hosts are routinely exploited bya community of parasite species [1ndash3] It is not surpris-ing therefore that organisms evolved elaborate responsestowards parasites under the selective pressure of multipleinfection [1 2 4 5] As the complexity of host-parasite andparasite-parasite interactions in natural multiparasite diseasesystems is not sufficiently described the principles governingthese interactions and host responses towards parasites are farfrom clear [6 7]

From an applied perspective identifying the health out-comes of coinfections involving helminthmacroparasites andmicroparasites such as protozoa bacteria or viruses is ofutmost importance to public and domestic animal healthofficials [4 8] This is on one hand because macroparasitecontrol has become widely available and efficient for human

populations and domestic animals On the other hand beforeany such macroparasite control programme is incorporatedin the fight against microparasites its potential effects on thefocal microparasites should be carefully taken into accountFor example despite intensive research it is still controversialwhether deworming in humans leads to decrease or increasein malaria incidence or severity [8]

Current theoretical and empiric work suggests that theoutcome of coinfection can be different even for the sameparasite system (eg [9]) depending on the level of resourcecompetition between macro- and microparasites [4] ormacroparasite burden and pathogenicity [2] The optimalhost immune response towards microparasites evolved notonly under a bias of the immune response towards chronicallyinfective macroparasites [10 11] but also under fluctuatingphysiological states and parasite burdens [6] Although host-parasite interactions have been intensively studied using

Hindawi Publishing Corporatione Scientific World JournalVolume 2014 Article ID 302903 6 pageshttpdxdoiorg1011552014302903

2 The Scientific World Journal

domestic and laboratory animals [5 12] it is questionablehow relevant these studies are for understanding how free-living mammals respond to concurrent macro- and micro-parasite infection [6 7 13 14]

Here we study the effect of anthelmintic treatment oncoccidia infection intensity in the free-living Alpine marmotM marmota latirostris Alpine marmots are commonly coin-fected with a tapeworm Ctenotaenia marmotae and coccidiaEimeria spp [15ndash18] While Eimeria spp persist in marmotsduring hibernation C marmotae is expulsed by marmotsbefore hibernation and reinfects them in the subsequentspring [16] Our study has two goals First we determinethe seasonal pattern in the infection intensity for the twocommonest endoparasites of the Alpine marmot Second wetest the hypotheses on the effect of helminth infection onmicroparasite infection intensity If helminths affect micro-parasites via a reduction in the hostrsquos immune response[4] we predict that the coccidia oocyst shedding rate formarmots exposed to anthelmintic treatment should be lowerthan that for marmots from the control group Alternativelyif helminths affect microparasites via a reduction in thehostrsquos resource levels [4] we predict that the coccidia oocystshedding rate formarmots exposed to anthelmintic treatmentshould be higher than that for marmots from the controlgroup

2 Materials and Methods

21 Study System We investigated the endoparasites of theAlpinemarmotMmarmota latirostris in an alpine grasslandof the Great Cold Valley in the High Tatra Mountains(49∘101015840N 20∘091015840E) Slovakia in 2011 The study site (ca200 ha) consists of about 16 territories (the number fluctuatesbetween years) of a marmot subpopulation that is underinvestigation since 2010The basic habitat features of the sub-population studied are presented in [19] The Alpine marmotis a true hibernating territorial rodent (Rodentia Sciuridae)living in social units of 2ndash20 individuals [20] The activeseason of the Alpine marmots in the study population inthe High Tatras spans from approximately mid-April to thebeginning of October (Radovan Vaclav unpublished data)

An anoplocephalid cestode Ctenotaenia (Cittotaenia)marmotae (Platyhelminthes Cestoda) is the most prevalentand abundant helminth endoparasite of the Alpine marmotacross its whole range including Slovakia [15 16 18 21ndash23] Tapeworms are composed of successive reproductivelyviable segments proglottids After maturation proglottidscontaining embryonated eggs detach from the tapeworm andare shed with host faeces The weight of the cestode canreach more than 3 of a marmotrsquos total body mass [17]C marmotae is expulsed from the marmot gastrointestinaltract before hibernation in autumn and the parasite recol-onizes its definite host in the subsequent spring [15 16]A variety of oribatid mite species act as intermediate hostsfor the tapeworm [24] with marmots acquiring infection byaccidentally ingesting infected mites [18]

Coccidia from the genus Eimeria are the most commonmicroparasites of the Alpine marmot [15 16] In contrastto C marmotae they do not require intermediate hosts for

transmission and persist inmarmots even during hibernation[17]Moreover in contrast toCmarmotae for which the peakin the proglottid shedding rate takes place in late summerthe peak in the coccidia oocyst shedding rate occurs inearly spring [17] It is assumed that the number of oocystsdischarged and the length of time they are shed depend onthe number of sporulated oocysts in the initial infective dose(eg [25]) If the host becomes exposed again to the samecoccidia the hostrsquos protective immunity is responsible for areduced oocysts shedding rate [25]

22 Experimental Design and Data Collection In spring2011 (late April-mid-May) we captured Alpine marmotsin two-door live-capture traps following a procedure byCohas et al [26] After capture marmots were tranquillizedwith Zoletil 100 (010mLkg) and marked with a numberedear tag and a transponder In order to address the effectof helminth macroparasite infection on coccidia micropar-asites each marmot was administered a broad-spectrumanthelmintic drug combination consisting of Ivermectinumand Praziquantelum A broad-spectrum anthelmintic treat-ment was used to eliminate the potential effects of infectionon coccidia not only by the most prevalent tapeworms butalso by any helminth taxa known to infect Alpine mar-mots [18] We administered 0025mLkg of Ivermectinum(Biomectin 10mgmL) and 01mLkg of Praziquantelum(Bancid 568mgmL) intramuscularly to each marmot fromthe experimental group and 0125mLkg of physiologicalsolution intramuscularly to each marmot from the controlgroupThe treatment was only applied once for eachmarmotThe effect of the treatment was examined considering allsampling units (individual faecal samples) at the level ofmarmot territories with four and three territories beingassigned to the experimental (11 marmots) and control (13marmots) groups respectively

From mid-May we conducted intensive behaviouralobservations to confirm the structure of marmot socialunits within territories and started to collect fresh marmotfaeces at seven territories where all (24) marmots werecaptured and either anthelmintic or physiological solutionhad been administered to them Forty-five faecal sampleswere collected during field visits in May July and Septemberat the specific defecation sites latrines which were usedexclusively by territory members For this study we did notinclude faeces collected frommarmot pastures because it wasnot possible to assign them unambiguously due to territoryoverlaps to individual territories Also juvenile marmotswhich normally emerge during July do not use latrinesand defecate freely within territories (personal observations)Therefore the faeces of untreated juveniles which also can beclassified visually based on their size were not included in ouranalysis

Fresh faecal samples were weighed to the nearest 01 g andstored in vials containing 25 (wv) aqueous K

2Cr2O7 The

vials were stored at ambient temperature for five days to allowoocyst sporulation while the content of each vial was mixedthoroughly twice every day After five days the aqueouscontent of vials was extracted into clean vials and stored ina cool place and then in the fridge until examination (within

The Scientific World Journal 3

3 months) Oocysts and tapeworm eggs and proglottids wereisolated by flotation in saturated sucrose solution and identi-fied using oil immersion lenses on a compound microscopeand Nomarski differential interference contrast microscopyThe oocyst numbers were quantified using the FLOTACtechnique Because most tapeworm proglottids containingeggs were crushed and did not allow exact quantification therate of tapeworm proglottid shedding was assigned to fourlevels at the scale from 0 to 3 based on the number of unam-biguously identified C marmotae proglottids All coprolog-ical samples were analysed at the University of Veterinaryand Pharmaceutical Sciences in Brno the Czech Republic

23 Data Analysis The effect of anthelmintic treatment onC marmotae and Eimeria spp shedding rates was examinedwith general linear mixed models (GLMM) Both responsevariables C marmotae proglottid shedding rate and log-transformed Eimeria spp oocyst numbers per gram (OPG)and faecal sample were analysed assuming normally dis-tributed errors A visual inspection of model validationgraphs (residuals versus fitted values and Q-Q plot) did notsuggest a clear violation of the assumptions of homogeneityand normality for neithermodel [27] Bothmodels containedtreatmentmonth and the interaction between treatment andmonth as fixed factors The tapeworm proglottid sheddingrate was used as a covariate in the model on Eimeria sppoocyst shedding Because samples collected from the sameterritories are not statistically independent territory identitywas entered as a random factor All tests were conducted withR software [28] using the lme4 package and the lmer function[29]The Type III tests with denominator degrees of freedomwere calculated based on a Satterthwaitersquos approximation andthe least-square means were calculated using the lmerTestpackage [30]The slice tests examining the equality of simpleeffects of one factor for a given level of the other factor werecalculated using the lsmeans package and the glht function[31] with the 119875 values reported being adjusted for multiplecomparisons using the Bonferroni single-step procedure

3 Results

31 Faecal Parasite Community Structure Examining 45marmot faecal samples collected from mid-May to the endof September at seven marmot territories the presence ofeggs or proglottids of a tapeworm Ctenotaenia marmotae(Platyhelminthes Anoplocephalidae) the eggs of a nema-tode Strongyloides sp (Nematoda Strongyloididae) and theoocysts of coccidia Eimeria spp (Apicomplexa Eimeriidae)was detected in 58 (2645 212 1423 and 1010 for MayJuly and September) 9 (445 112 323 and 010 for MayJuly and September) and 80 (3645 912 2023 and 710for May July and September) samples respectively

32 Effect of Anthelmintic Treatment on Tapeworm FaecalShedding Considering the whole active season of marmotsanthelmintic treatment conducted shortly after hibernationdid not significantly lower the total shedding rate ofCtenotae-nia marmotae (effect of treatment 119865

1 464= 024 119875 = 0644

Cten

otae

nia

mar

mot

ae sh

eddi

ng ra

te (plusmn

SE)

25

20

15

10

05

00

minus05May July September

lowast

lowastlowast

lowast

0078

Control groupExperimental group

Figure 1 Effect of anthelmintic treatment on the faecal sheddingrate of Ctenotaenia marmotae proglottids in the Alpine marmotM marmota latirostris The C marmotae shedding rate representsproglottid numbers on the scale from 0 to 3 per gram of afaecal sample as more precise proglottidegg quantification was notpossible due to the physical damage ofmost proglottidsThe symbolsabove lines lowastlowast lowast and 0078 refer to 119875 lt 0001 119875 lt 005 and119875 = 0078 respectively The 119875 values reported are adjusted formultiple comparisons using the Bonferroni single-step procedure

effect of time of season 1198652 3878= 883 119875 lt 0001 and

interaction between treatment and time of season 1198652 3887=

112 119875 = 0333) Yet examination of the interaction betweenexperimental treatment and time of season revealed thatanthelmintic treatment affected the dynamics of tapewormfaecal shedding during specific periodsNamely the sheddingrate increased from July to September but only for marmotsin the control group (effect of time of season sliced by exper-imental treatment difference between July and Septemberfor control group 119875 = 0012 difference between July andSeptember for experimental group119875 = 0637 Figure 1) Alsothe proglottid shedding rate tended to increase from May toJuly but only for marmots in the experimental group (effectof time of season sliced by experimental treatment differencebetweenMay and July for control group119875 = 0631 differencebetween May and July for experimental group 119875 = 0078Figure 1) That is to say while the tapeworm shedding ratefor the control group increased exponentially from May toSeptember the tapeworm shedding rate for the experimentalgroup was sigmoidal increasing abruptly from May to Julyand remaining high thereafter

33 Effect of Anthelmintic Treatment on Coccidia Faecal Shed-ding While the shedding rate of tapeworm proglottids andEimeria spp oocysts was negatively associated (estimate plusmnSE = ndash102 plusmn 049 119905

3712= ndash207 119875 = 0045) we did not find a

significant difference in the shedding rate of oocysts betweenanthelmintic and control groups (effect of treatment119865

1 562=

0452 119875 = 0528 effect of time of season 1198652 3646= 348


May July September

lowast

Eim

eria

spp

shed

ding

rate

(plusmnSE

)

8

7

6

5

4

3

2

1

0


Figure 2 Effect of anthelmintic treatment on the faecal sheddingrate of Eimeria spp oocysts in the Alpine marmot M mar-mota latirostris The Eimeria spp shedding rate represents log-transformed oocyst numbers per gram of a faecal sample Thesymbol above line lowast refers to 119875 lt 005 The 119875 value reported isadjusted for multiple comparisons using the Bonferroni single-stepprocedure

119875 = 0041 and interaction between treatment and time ofseason 119865

2 3751= 1417 119875 = 0255) Nevertheless similarly

as for the dynamics of tapeworm shedding we found that theshedding rate of Eimeria spp oocysts increased from May toJuly but only for marmots in the experimental group (effectof time of season sliced by experimental treatment differencebetweenMay and July for control group119875 = 0858 differencebetween May and July for experimental group 119875 = 0036Figure 2)

4 Discussion

Our experimental results suggest that helminth infectionhas a protective effect on the reproductive rates of coc-cidia microparasites in free-living Alpine marmots (hereafterreferred to as marmots) Specifically we found that whenhelminth infection was postponed in the beginning of themarmot active season via anthelmintic treatment an increasein the shedding rate of Eimeria spp oocysts also was post-poned Interestingly a correlative result revealed a negativeassociation between the infection intensity of a tapewormmacroparasite C marmotae and the coccidia microparasitesof the genus Eimeria indicating that under increased hostexploitation tapeworms negatively affect coccidia reproduc-tive rates

We stress that our deworming treatment affected onlyexisting helminth infections because the shedding rates oftapeworm proglottids were comparable between experimen-tal and controlmarmot groups from themid active season (cf[32]) Thus our treatment can be considered as a postpone-ment rather than prevention of helminth infection and larger

sample size would unlikely lead to revealing a significanteffect of deworming treatment on tapeworm infection duringthewhole season Importantly this study shows thatmarmotsof the study population in the High Tatras started to shed theproglottids of C marmotae already in the second half of MayAssuming the prepatent period of C marmotae to be 39ndash50days [17] this study indicates thatmarmots can acquire infec-tions with C marmotae already during early April that isafter hibernation but before emergence This finding sug-gesting that the first tapeworm infections take place in theunderground burrow system is important because themech-anism of host infections with Anoplocephalid tapeworms isstill unclear [19]

Similar to previous studies conducted elsewhere [15ndash17]we found that Eimeria spp prevalence is high in marmots inthe High Tatras However compared to a previous study byCallait et al [17] which revealed the peak in coccidia oocystshedding during early spring we found that the Eimeria sppoocyst shedding rates in control marmots remained highthrough summer It is possible that this difference is dueto higher coccidia diversity in the High Tatras and a corre-spondingly longer time taking marmots to acquire resistanceagainst this complex parasite group In addition this patterncan be due to a longer time takingmarmots in theHighTatrasto accumulate or sustain growing helminths thereby allowingcoccidia to reproduce for a longer timewithout elevated com-petition for host resourcesThe importance of coccidia diver-sity and seasonal changes in tapeworm burdens warrantsfurther studies

Our work lends support to the hypothesis [4] thathelminth infection has a protective effect on concurrentmicroparasite infections via modulated host immunity Nev-ertheless our work also points to the fact that within-hostinteractions between helminths and microparasites can becontext-dependent [2] and they can change to negative onesonce helminth parasites mature and start to reproduce Thisis feasible in our study system because both tapeworms Cmarmotae and coccidia Eimeria spp share the same bodysections of their marmot hosts

Based on our results we suggest that coccidia benefitfrom intermittent helminth infection in marmots due to theprotective effects of helminth infection during the first phaseof the host active season In turn from the host perspectivethis study indicates that marmot immune response towardscontinuous coccidia infection would be optimal only underno helminth infection when the host immune responseis not compromised by a downregulated Th1-like response(eg [4 5]) Even if this situation is hardly possible undernatural conditions we propose that intestinal worm expul-sion by marmots regularly taking place just before hiberna-tion might represent not only intestinal self-cure of worminfections [15 17] but also an optimal strategy to controlmicroparasite infection intensity during an important partof the host life cycle shortly after hibernation

Coinfection withmultiple parasites or parasite strains hasan important bearing on parasite virulence because undercompetition parasites are thought to increase host resourceexploitation [7] Our study suggests that coccidia virulencemight not be heightened in marmot populations if tapeworm


infections are low or occur later in the season because compe-tition between the two parasites groups is likely to be low (cf[33]) In contrast Eimeria virulence can be higher in seasonsor marmot populations exhibiting higher worm burdenor earlier worm infections when interspecific competitionamong parasites is likely to be elevated Finally interstraincompetition also can affect parasite virulence in marmotsbecause we detected a complex of distinct morphotypes ofEimeria spp in faecal samples A future work needs to estab-lish whether specific Eimeria morphotypes prevail underdifferent tapeworm burdens

5 Conclusion

This experimental work demonstrates that correlational stud-ies as well as those studies limiting their scope to macro-parasite prevalence are not sufficient to depict the complex-ity of macroparasite-microparasite interactions Our resultssuggest that the synergic effects of tapeworm infection onEimeria spp microparasites change into antagonistic onesunder increased host exploitation by grown and reproducingmacroparasites We suggest that studying host-parasite andparasite-parasite interactions in free-ranging host speciessuch as marmots showing intermittent helminth infec-tions would be rewarding because this system mimics theone involving human populations receiving intermittentanthelmintic treatment while concurrently fighting micro-parasite diseases such as malaria

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

Theauthors are indebted toDr DavidModry and his team forhelp with coccidia analysis Dr Martina Miterpakova helpedwith helminth identification Four anonymous reviewershelped to improve this paper Lrsquoudovıt Zahor and the staffof the Zbojnıcka chata cabin provided invaluable logisticsupport during the field season Dr Lrsquoubomır Vidlicka helpedwith sample logisticsThis studywas funded by aVEGAgrantof the Ministry of Education Science Research and Sport ofthe Slovak Republic (no 2017610)The study was conductedunder approval from the Ministry of Environment of theSlovak Republic (no 49832010-211)

References

[1] F E G Cox ldquoConcomitant infections parasites and immuneresponsesrdquo Parasitology vol 122 supplement pp S23ndashS382001

[2] A Fenton ldquoDances with worms the ecological and evo-lutionary impacts of deworming on coinfecting pathogensrdquoParasitology vol 140 no 9 pp 1119ndash1132 2013

[3] T N Petney and R H Andrews ldquoMultiparasite communitiesin animals and humans frequency structure and pathogenic

significancerdquo International Journal for Parasitology vol 28 no3 pp 377ndash393 1998

[4] A L Graham ldquoEcological rules governing helminth-micro-parasite coinfectionrdquo Proceedings of the National Academy ofSciences of the United States of America vol 105 no 2 pp 566ndash570 2008

[5] E Moreau and A Chauvin ldquoImmunity against helminthsinteractions with the host and the intercurrent infectionsrdquo Jour-nal of Biomedicine and Biotechnology vol 2010 Article ID428593 9 pages 2010

[6] ZMWeil L BMartin II andR JNelson ldquoInteractions amongimmune endocrine and behavioural response to infectionrdquo inMicrommals and Microparasites From Evolutionary Ecology toManagement S Morand B R Krasnov and R Poulin Eds pp443ndash474 Springer Tokyo Japan 2006

[7] S Bordes and S Morand ldquoThe impact of multiple infections onwild animal hosts a reviewrdquo Infection Ecology amp Epidemiologyvol 1 Article ID 7346 2011

[8] M Nacher ldquoInteractions between worms and malaria goodworms or badwormsrdquoMalaria Journal vol 10 article 259 2011

[9] G Karadjian D Berrebi N Dogna et al ldquoCo-infectionrestrains Litomosoides sigmodontis filarial load and plasmodialP yoelii but not P chabaudi parasitaemia in micerdquo Parasite vol21 article 16 2014

[10] O O Barriga ldquoEvidence and mechanisms of immunosup-pression in tick infestationsrdquo Genetic Analysis BiomolecularEngineering vol 15 no 3ndash5 pp 139ndash142 1999

[11] J A Jackson I M Friberg S Little and J E Bradley ldquoReviewseries on helminths immune modulation and the hygienehypothesis immunity against helminths and immunologicalphenomena in modern human populations coevolutionarylegaciesrdquo Immunology vol 126 pp 18ndash27 2009

[12] W C Gause J F Urban Jr and M J Stadecker ldquoThe immuneresponse to parasitic helminths insights from murine modelsrdquoTrends in Immunology vol 24 no 5 pp 269ndash277 2003

[13] S C L Knowles A Fenton O L Petchey T R Jones R Barberand A B Pedersen ldquoStability of within-host-parasite commu-nities in a wild mammal systemrdquo Proceedings of the RoyalSociety B vol 280 no 1762 Article ID 20130598 2013

[14] A B Pedersen and J Antonovics ldquoAnthelmintic treatmentalters the parasite community in a wild mouse hostrdquo BiologyLetters vol 9 no 4 Article ID 20130205 2013

[15] B Bassano B Sabatier L Rossi and E Macchi ldquoParasitic faunaof the intestinal tracts of Marmota marmota in the westernAlpsrdquo in Proceedings of 1st International Symposium on AlpineMarmot and on GenusMarmota B Bassano P Durio U Gallo-Orsi and E Macchi Eds pp 13ndash24 Torino Italy 1992

[16] M P Callait and D Gauthier ldquoParasite adaptations to hiberna-tion in Alpine marmots (Marmota marmota)rdquo in Proceedings ofthe 11th International Hibernation Symposium Life in the ColdG Heldmaier and M Klingenspor Eds pp 139ndash146 SpringerJungholz Austria 2000

[17] M P Callait D Gauthier and C Prudrsquohomme ldquoAlpine mar-mots and their digestive parasites infection kinetic and para-sitic strategyrdquo in Holarctic Marmots As a Factor of BiodiversityAbstracts of the 3rd International Conference on Marmots K BArmitage andV Y Rumiantsev Eds pp 113ndash122 ABFMoscowRussia 1997

[18] M Preleuthner ldquoParasiten des Alpenmurmeltieres (Marmotamarmota) Systematik Entwicklung Verbreitungrdquo Stapfia vol146 pp 77ndash92 1999


[19] R Vaclav and S Kaluz ldquoThe effect of herbivore faeces on theedaphic mite community implications for tapeworm transmis-sionrdquo Experimental and Applied Acarology vol 62 no 3 pp377ndash390 2014

[20] C Perrin D Allaine and M Le Berre ldquoSocio-spatial organi-zation and activity distribution of the alpine marmotMarmotamarmota preliminary resultsrdquo Ethology vol 93 pp 21ndash30 1993

[21] C Gortazar J Herrero A Garcıa-Serrano J Lucientes and DF Luco ldquoPreliminary data on the parasitic fauna of the digestivesystem of Marmota marmota in the Western Pyreneesrdquo inBiodiversity in Marmots M Le Berre R Ramousse and LLe Guelte Eds pp 105ndash108 International Marmot NetworkMoscow Russia 1996

[22] M T Manfredi E Zanin and A P Rizzoli ldquoHelminth commu-nity on alpine marmotsrdquo in Proceedings of the 1st InternationalSymposium on AlpineMarmot and GenusMarmota B BassanoP Durio U Gallo-Orsi and E Macchi Eds pp 203ndash207Torino Italy 1992

[23] F Tenora ldquoDiscovery of a tapeworm Ctenotaenia marmotaeFrolich 1802Railliet 1893 in CSSRrdquo Foolia Zoologica vol 10 p396 1961

[24] E Ebermann ldquoOribatiden (Oribatei Acari) als Zwischenwirtedes Murmeltier-Bandwurmes Ctenotaenia marmotae (Frolich 1802)rdquo Parasitology Research vol 50 pp 303ndash312 1976

[25] D W Duszynski and S J Upton ldquoCyclospora EimeriaIsospora and Cryptosporidium spprdquo in Parasitic Diseases ofWildAnimalsWM SamuelM J Pybus andAA Kocan Edspp 416ndash459 Iowa State University Press Ames Iowa USA2001

[26] A Cohas N G Yoccoz C Bonenfant et al ldquoThe geneticsimilarity between pair members influences the frequency ofextrapair paternity in alpine marmotsrdquo Animal Behaviour vol76 no 1 pp 87ndash95 2008

[27] A F Zuur E N Ieno N J Walker A A Saveliev and G MSmith Mixed Effects Models and Extensions in Ecology with RSpringer New York NY USA 2009

[28] R Core Team R A Language and Environment for StatisticalComputing R Foundation for Statistical Computing ViennaAustria 2014 httpwwwR-projectorg

[29] D Bates M Maechler B Bolker and S Walker ldquolme4 Linearmixed-effects models using Eigen and S4 R package version 11-6rdquo 2014 httpCRANR-projectorgpackage=lme4

[30] A Kuznetsova P B Brockhoff and R H B ChristensenldquolmerTest Tests for random and fixed effects for linear mixedeffect models (lmer objects of lme4 package) R package version20-6rdquo 2014 httpCRANR-projectorgpackage=lmerTest

[31] R V Lenth ldquolsmeans Least-Squares Means R package version200-5rdquo 2014 httpCRANR-projectorgpackage

[32] M P Callait ldquoMarmotte alpine et endoparasites biologie desparasites et comparaison delinfestation sur differents sitesrdquo inProceedings of the 4eme Journee dEtude sur la Marmotte AlpineR Ramousse and M Le Berre Eds pp 53ndash58 InternationalMarmot Network Lyon France 1997

[33] S Schjoslashrring and J C Koella ldquoSub-lethal effects of pathogenscan lead to the evolution of lower virulence in multiple infec-tionsrdquo Proceedings of the Royal Society B Biological Sciences vol270 pp 189ndash193 2003

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


domestic and laboratory animals [5 12] it is questionablehow relevant these studies are for understanding how free-living mammals respond to concurrent macro- and micro-parasite infection [6 7 13 14]

Here we study the effect of anthelmintic treatment oncoccidia infection intensity in the free-living Alpine marmotM marmota latirostris Alpine marmots are commonly coin-fected with a tapeworm Ctenotaenia marmotae and coccidiaEimeria spp [15ndash18] While Eimeria spp persist in marmotsduring hibernation C marmotae is expulsed by marmotsbefore hibernation and reinfects them in the subsequentspring [16] Our study has two goals First we determinethe seasonal pattern in the infection intensity for the twocommonest endoparasites of the Alpine marmot Second wetest the hypotheses on the effect of helminth infection onmicroparasite infection intensity If helminths affect micro-parasites via a reduction in the hostrsquos immune response[4] we predict that the coccidia oocyst shedding rate formarmots exposed to anthelmintic treatment should be lowerthan that for marmots from the control group Alternativelyif helminths affect microparasites via a reduction in thehostrsquos resource levels [4] we predict that the coccidia oocystshedding rate formarmots exposed to anthelmintic treatmentshould be higher than that for marmots from the controlgroup

2 Materials and Methods

21 Study System We investigated the endoparasites of theAlpinemarmotMmarmota latirostris in an alpine grasslandof the Great Cold Valley in the High Tatra Mountains(49∘101015840N 20∘091015840E) Slovakia in 2011 The study site (ca200 ha) consists of about 16 territories (the number fluctuatesbetween years) of a marmot subpopulation that is underinvestigation since 2010The basic habitat features of the sub-population studied are presented in [19] The Alpine marmotis a true hibernating territorial rodent (Rodentia Sciuridae)living in social units of 2ndash20 individuals [20] The activeseason of the Alpine marmots in the study population inthe High Tatras spans from approximately mid-April to thebeginning of October (Radovan Vaclav unpublished data)

An anoplocephalid cestode Ctenotaenia (Cittotaenia)marmotae (Platyhelminthes Cestoda) is the most prevalentand abundant helminth endoparasite of the Alpine marmotacross its whole range including Slovakia [15 16 18 21ndash23] Tapeworms are composed of successive reproductivelyviable segments proglottids After maturation proglottidscontaining embryonated eggs detach from the tapeworm andare shed with host faeces The weight of the cestode canreach more than 3 of a marmotrsquos total body mass [17]C marmotae is expulsed from the marmot gastrointestinaltract before hibernation in autumn and the parasite recol-onizes its definite host in the subsequent spring [15 16]A variety of oribatid mite species act as intermediate hostsfor the tapeworm [24] with marmots acquiring infection byaccidentally ingesting infected mites [18]

Coccidia from the genus Eimeria are the most commonmicroparasites of the Alpine marmot [15 16] In contrastto C marmotae they do not require intermediate hosts for

transmission and persist inmarmots even during hibernation[17]Moreover in contrast toCmarmotae for which the peakin the proglottid shedding rate takes place in late summerthe peak in the coccidia oocyst shedding rate occurs inearly spring [17] It is assumed that the number of oocystsdischarged and the length of time they are shed depend onthe number of sporulated oocysts in the initial infective dose(eg [25]) If the host becomes exposed again to the samecoccidia the hostrsquos protective immunity is responsible for areduced oocysts shedding rate [25]

22 Experimental Design and Data Collection In spring2011 (late April-mid-May) we captured Alpine marmotsin two-door live-capture traps following a procedure byCohas et al [26] After capture marmots were tranquillizedwith Zoletil 100 (010mLkg) and marked with a numberedear tag and a transponder In order to address the effectof helminth macroparasite infection on coccidia micropar-asites each marmot was administered a broad-spectrumanthelmintic drug combination consisting of Ivermectinumand Praziquantelum A broad-spectrum anthelmintic treat-ment was used to eliminate the potential effects of infectionon coccidia not only by the most prevalent tapeworms butalso by any helminth taxa known to infect Alpine mar-mots [18] We administered 0025mLkg of Ivermectinum(Biomectin 10mgmL) and 01mLkg of Praziquantelum(Bancid 568mgmL) intramuscularly to each marmot fromthe experimental group and 0125mLkg of physiologicalsolution intramuscularly to each marmot from the controlgroupThe treatment was only applied once for eachmarmotThe effect of the treatment was examined considering allsampling units (individual faecal samples) at the level ofmarmot territories with four and three territories beingassigned to the experimental (11 marmots) and control (13marmots) groups respectively

From mid-May we conducted intensive behaviouralobservations to confirm the structure of marmot socialunits within territories and started to collect fresh marmotfaeces at seven territories where all (24) marmots werecaptured and either anthelmintic or physiological solutionhad been administered to them Forty-five faecal sampleswere collected during field visits in May July and Septemberat the specific defecation sites latrines which were usedexclusively by territory members For this study we did notinclude faeces collected frommarmot pastures because it wasnot possible to assign them unambiguously due to territoryoverlaps to individual territories Also juvenile marmotswhich normally emerge during July do not use latrinesand defecate freely within territories (personal observations)Therefore the faeces of untreated juveniles which also can beclassified visually based on their size were not included in ouranalysis

Fresh faecal samples were weighed to the nearest 01 g andstored in vials containing 25 (wv) aqueous K

2Cr2O7 The

vials were stored at ambient temperature for five days to allowoocyst sporulation while the content of each vial was mixedthoroughly twice every day After five days the aqueouscontent of vials was extracted into clean vials and stored ina cool place and then in the fridge until examination (within




3 Results



1 464= 024 119875 = 0644

Cten

otae

nia

mar

mot

ae sh

eddi

ng ra

te (plusmn

SE)

25

20

15

10

05

00


lowast

lowastlowast

lowast

0078









1 562=



May July September

lowast

Eim

eria

spp

shed

ding

rate

(plusmnSE

)

8

7

6

5

4

3

2

1

0






4 Discussion










5 Conclusion




Acknowledgments


References











































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




3 Results



1 464= 024 119875 = 0644

Cten

otae

nia

mar

mot

ae sh

eddi

ng ra

te (plusmn

SE)

25

20

15

10

05

00


lowast

lowastlowast

lowast

0078









1 562=



May July September

lowast

Eim

eria

spp

shed

ding

rate

(plusmnSE

)

8

7

6

5

4

3

2

1

0






4 Discussion










5 Conclusion




Acknowledgments


References











































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


May July September

lowast

Eim

eria

spp

shed

ding

rate

(plusmnSE

)

8

7

6

5

4

3

2

1

0






4 Discussion










5 Conclusion




Acknowledgments


References











































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



5 Conclusion




Acknowledgments


References











































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology
























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Research Article The Effect of Anthelmintic Treatment on...

Documents

Transcript of Research Article The Effect of Anthelmintic Treatment on...