Research ArticleA Long Temporal Study of Parasitism in Asexual-SexualPopulations of Carassius gibelio Does the Parasite InfectionSupport Coevolutionary Red Queen Dynamics

Tomaacuteš Pakosta1 Lukaacuteš Vetešniacutek2 and Andrea Šimkovaacute 1

1Department of Botany and Zoology Faculty of Science Masaryk University Kotlarska 2 611 37 Brno Czech Republic2Institute of Vertebrate Biology Academy of Sciences of the Czech Republic vvi Kvetna 8 603 65 Brno Czech Republic

Correspondence should be addressed to Andrea Simkova simkovascimunicz

Received 15 December 2017 Accepted 11 February 2018 Published 11 March 2018

Academic Editor Yann Quilichini

Copyright copy 2018 Tomas Pakosta et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Carassius gibelio is an extraordinary cyprinid species exhibiting both sexual and asexual reproduction We hypothesized thatparasitism selection is one of the potential mechanisms contributing to the coexistence of the two reproductive forms of C gibelioliving in the same habitat We performed a four-year study to investigate the dynamics of parasite infection in C gibelio Accordingto the Red Queen prediction the asexual form is a target of parasite adaptation due to its low genetic variability Both sexualand gynogenetic forms of C gibelio exhibited similar levels of prevalence with monogeneans being the most frequently observedparasite group We observed the temporal dynamics of parasite infection in the last year of investigation when both forms weremore strongly parasitized The sexual form was more parasitized by ectoparasites in the first and last years and less parasitized bynematodes in the last year when compared to the gynogenetic form We found no trend of high parasite infection in gynogeneticmtDNA haplotypes We conclude that Red Queen dynamics is not the mechanism driving parasite infection in sexual-gynogeneticC gibelio over a long time scale Alternatively we suggest that the dynamics of parasite infection in this complex may be generatedby multiple mechanisms

1 Introduction

Thecoexistence of sexual and asexual forms is rarely reportedin vertebrates The stable coexistence of these two formsmay be achieved if there are some disadvantages for asexualscompensating the evolutionary twofold costs of sexual repro-duction [1 2] The coexistence of asexual and sexual repro-duction is documented only in a small number of freshwaterfishes including nine genera [3] In the context of asexualreproduction gynogenesis has a specific constraint as thesperm of males are necessary to induce embryogenesis thatis the eggs of gynogenetic females are stimulated by sperm ofusually conspecific (or alternatively phylogenetically closelyrelated) males to start embryogenesis but the sperm do notfuse with the egg nucleus Thus in such case the asexual andsexual forms are forced to coexist as the persistence of thesexual form is in the direct interest of the gynogenetic form

This means that gynogenetic form (which may be consideredas a ldquoparasiterdquo using the sperm of sexual males) cannotoutcompete the sexual form (which may be considered asldquohostrdquo for gynogenetic form)

The Carassius auratus complex is one of the rare com-plexes exhibiting both gynogenetic and sexual reproductionThis nonnative species complex colonized Europe in thesecond half of the 20th century and is considered to bethe most successful invasive species in European waters [4ndash9] very effectively expanding and adapting to new habitatsdue to its high ecological tolerance [10] Representativesof the C auratus complex entered the Czech hydrologicalsystem by means of migration from the Danube in around1975 [7 11 12] and over the next 20 years the complexspread to the Danube tributaries that is the Morava andDyje Rivers where it produced stable populations [13] Thefirst populations of the C auratus complex were triploid

HindawiBioMed Research InternationalVolume 2018 Article ID 6983740 10 pageshttpsdoiorg10115520186983740

2 BioMed Research International

gynogenetic females [14 15] Fifteen years after the ini-tial invasion however mixed diploid-polyploid populationsincluding sexual diploid males and females gynogenetictriploid females and rarely triploid males or even tetraploidspecimens were reported [16ndash18] In recent times bothreproductive forms have coexisted successfully in the samehabitats The particular success of colonization by formerunisexual gibel carp populations was the result mainly ofgibel carprsquos specific biological properties that is the abilityto reproduce gynogenetically resulting in high reproductiveoutput but was also due to its own invasive activitiesfavourable environmental factors and human activity [14] Itseems that the shift from gynogenesis to sexual reproductionfollowed the stabilization of C gibelio populations in the suc-cessfully colonized habitats In the area of theCzechRepublicthe C auratus complex is represented by four mitochondriallineagesC gibelioC auratusC langsdorfii and the so-calledM line Carassius gibelio is the most commonly occurringform of this complex in the Czech Republic [19]

Several mechanisms have been studied and suggested ascontributing to the coexistence of sexual and asexual formsin fish complexes Some of them have also been hypothesizedand examined with respect to the C auratus complex Therole of parasites in maintaining the coexistence of asexualand sexual reproduction is based on the Red Queen (RQ)hypothesis which predicts antagonistic coevolutionary host-parasite interactions leading to sustained oscillations in theirgenotype frequencies [20 21] Following the RQ hypothesisthe asexual form is due to its low genetic recombination atarget of parasite adaptation [22] as parasites adapt quicklyto common genotypes [23] In contrast the sexual formquickly and effectively produces combinations of parasite-resistant genes and therefore is able to escape infection [2224] In accordance with the RQ hypothesis Hakoyama etal [25] showed that parasitism plays an important role inmaintaining the coexistence of sexual and gynogenetic formsof the C auratus complex They reported higher prevalenceof the trematode species Metagonimus sp in gynogeneticfemales when compared to diploid females in natural habi-tats In accordance with this observation Hakoyama andIwasa [26] concluded that the coexistence of sexual andgynogenetic forms of the C auratus complex is stabilizedby parasites However the mechanisms predicted by the RQhypothesis did not explain the coexistence of sexual andasexual forms in other freshwater fish complex that isCobitistaenia hybrid complex [27] In that case differential habitatpreferences proposed by niche shift hypothesis contributedto the coexistence of both forms In addition Hakoyamaand Iwasa [26] also proposed that the coexistence of Cauratus complex is not strictly maintained by parasites butmay result from interactions between several factors (ieassortative mating female-biased sex allocation differentdispersion capacities and different utilization of niches)Simkova et al [28] examined the parasite load in gynogeneticand sexual C gibelio in the light of the RQ hypothesis Theyapplied the genotyping of major histocompatibility complex(MHC) IIB genes and showed that the most common MHCgenotype of the gynogenetic form is more parasitized thansexual genotypes or rare gynogenetic genotypes In addition

Simkova et al [29] hypothesized that the different invest-ments in condition- growth- and fitness-related traits mayrepresent other mechanisms contributing to the coexistenceof gynogenetic and sexual forms of the C auratus complexHowever they revealed similar growth expressed in terms ofbody size and similar condition factors in gynogenetic andsexual forms and failed to identify the reproductive disad-vantage for the gynogenetic form when investment in repro-duction was measured by gonad weight and estradiol levelThis indicates that similar energy is invested in the reproduc-tion of gynogenetic and sexual females In addition Simkovaet al [29] proposed the existence of a new mechanismpotentially contributing to the coexistence of gynogeneticand sexual forms of the C auratus complex that is thelow aerobic performance in gynogens which may repre-sent a physiological disadvantage balancing the evolutionarycost of sexual reproduction In addition Vetesnık et al[30] identified higher concentrations of triacylglycerols andcholesterol in the blood plasma of gynogenetic femalesindicating a higher metabolic rate and higher energy intakewhen compared to sexuals

Finally the role of immunity in maintaining the coex-istence of sexual and gynogenetic forms of the C auratuscomplex was examined While Hakoyama et al [25] showedlower nonspecific immunity and higher parasite loads ofnonspecific trematode parasite species (Metagonimus sp)in the gynogenetic form of the C auratus complex (theirstudy was performed using gynogenetic C langsdorfii andan unspecified sexual line of C auratus complex) Simkovaet al [28 29] showed no difference in nonspecific immunitybetween the gynogenetic and sexual forms of C gibeliobut higher specific immunity (IgM level) as well as higherparasite loads of host specific monogeneans (Dactylogyrusspp) in the gynogenetic form when compared to the sexualform

As the pattern of parasite infection in fish populationscomposed of asexual and sexual forms had not yet beenstudied over a long time scale we focused this study onthe temporal dynamics of parasite infection The aim wasto perform a long-term study (over four consecutive years)to investigate the composition of parasite communities andparasite load in asexual and sexual forms of C gibelio

2 Materials and Methods

21 Fish Sampling A total of 203 individuals of the Carassiusauratus complex (standard length 2835 plusmn 261 cm) werecaught by electrofishing in the River Dyje near the city ofBreclav (48∘801015840N 16∘841015840E theMorava River basin) Fishweresampled during the same period in each of four consecutiveyears (first two weeks in August 2012ndash2015) After bloodand fin sampling fish were immediately placed in a tankcontaining original water and transported to the laboratoryFor each specimen the total and standard lengths bodyweight and sex were recorded The complete parasitologicaldissection of fish was performed following Ergens and Lom[31] A fin clip of each specimenwas preserved in 90 ethanolfor molecular analyses

BioMed Research International 3

22 Parasite Collection and Identification Monogeneanswere removed from the gills or fins placed into a drop ofwater on a slide coveredwith a coverslip andfixedusing glyc-erine ammonium picrate (GAP) for later analysis [32] Theremaining parasite species were fixed in 70 ethanol Parasitespecimens were examined using an Olympus model BX50light microscope (including an Olympus DP71 digital micro-scope camera) with phase contrast differential interferencecontrast and digital imaging (MicroImage 40 forWindows)

23 Ploidy Determination and Molecular Analyses A fin clipof about 1 cm2 in area was taken from each fish for ploidydetection and fixed in 70 ethanol Before analysis this tissuewas homogenised on a Petri dish in a 2ml solution of CyStainDNA 1 step Partec and relative DNA content was estimatedusing a Partec CCA I continuous flow cytometer (PartecGmbH httpswwwsysmex-parteccom) as was applied pre-viously for ploidymeasurements using blood sample [33 34]Fresh blood of diploid C auratus was obtained by punctureof the caudal blood vessel using a heparinised syringe andwas used as a reference standard Overall for our study 112diploid specimens (comprising 56 females and 56 males)and 91 triploid females were randomly selected The controlregion ofmitochondrial DNA (D-loop) was analysed for eachspecimen DNA was extracted by Dneasy Blood and TissueKit (Qiagen) from fin clips preserved in ethanol PCR andDNA sequencing were performed according to Sambrooket al [35] with small modifications following Vetesnık et al[36] and Papousek et al [37] For amplification the forwardprimer CarU32 (51015840-CCAAAGCCAGAATTCTAAAC-31015840) andthe reverse primer CarL509 (51015840-CATGTGGGGTAATGA-31015840)[37] were used A PCRproduct of 475ndash493 bp in length coversall ETAS domains (representing a high degree of variabilityconsidered useful for the study of intra- and interspeciesevolution) [38] and a part of the central domain in thecontrol region PCR was performed in a final volume of 30 120583lcontaining 1x PCR reaction buffer 15mMMgCl

2 03 120583M of

each primer (CarU32 and CarL509) 02mM dNTPs 15 UTaq polymerase and 100 ng120583l of template gDNA PCR wascarried out using the following steps 1min of denaturationat 95∘C followed by 32 cycles of 45 s at 94∘C 30 s at 50∘Cand 45 s at 72∘C and 10min of final elongation at 72∘C ThePCR products were checked on 15 agarose gel purified byHigh Pure PCR product purification kit (Roche) and directlysequenced using BigDye Terminator Cycle sequencing kit onan ABI 3130 Genetic Analyser (Applied Biosystems USA)The sequences were aligned using MEGA 60 [39] andanalysed using TCS 121 [40] The obtained sequences werecompared with the sequences for the mtDNA haplotypes ofthe C auratus complex available in GenBank The sequencesof mtDNA haplotypes obtained in this study were identicalwith the sequences available in GenBank under the followingaccession numbers FJ167410 FJ167411 FJ167413 FJ167414FJ167416 FJ167420 and FJ167423ndashFJ167425

24 Statistical Analyses General linear models (GLM) wereused to evaluate the threemain effects that is ploidy sex andyear on parasite abundance as well as two interaction effectsthat is ploidy with year and ploidy with body size The

standard body length of fish was included as a covariateParasite abundance was expressed by the following variablestotal parasite abundance (ie the abundance including allparasite specimens) total monogenean abundance (ie theabundance of the most common parasitic group) totalabundance of Nematoda (ie the abundance of the mostcommon endoparasite group) and the abundance of themost common parasitic species or genera that is Dactylo-gyrus spp (gill Monogenea) Gyrodactylus spp (gill and finMonogenea) and Ichthyophthirius multifiliis (Protozoa) Formultiple comparisons the Tukey post hoc test was used Allvariables applied in this study were examined for normaldistribution and homogeneity of variance For data that didnot fit these assumptions log-transformation was appliedStatistical analyses were performed in Statistica for Windows12 StatSoft Inc

3 Results

Random sampling of the mixed population of C gibelio inthe studied locality over a period of four years revealedthe following composition of the mixed gynogenetic-sexualpopulation 38 of fish were triploid specimens (37 weregynogenetic females and 1 were triploid males) and 63of fish were sexual diploid specimens Molecular analysis ofmtDNA confirmed that all specimens were representativesof the C auratus complex Our study revealed the presenceof 9 variants of 16 mtDNA haplotypes previously describedin the Southern Moravian region of the Czech Republic byPapousek [19] (Table 1) The Carassius gibelio lineage was themost frequent in the studied locality (96) Six mtDNAhaplotype variants of C gibelio were identified G01 G02G04 G05 G07 and G11 with G02 representing the mostcommon haplotype (Table 1) Two of the other haplotypesG01 and G04 were also common For sexual specimens G02and G04 were the most common haplotypes and G05 was araremitochondrial genotype in all four years of investigationFor gynogenetic females differences in haplotype frequenciesamong the four years were observed The most frequenthaplotypes of gynogenetic females reported in each yearwere as follows G01 G02 and G07 in 2012 G02 in 2013and G01 in both 2014 and 2015 Among the 6 C gibeliomtDNA haplotypes 3 haplotypes were found exclusively ingynogenetic females one was found exclusively in sexualspecimens and two were shared by sexual and gynogeneticspecimens The other three lineages C auratus (haplotypeA01) C langsdorfii (haplotype L01) and M line (haplotypeM01) were rare (24 05 and 1 resp) For the followinganalyses only the C gibelio lineage was selected due to thehigh frequency of this form

Parasite communities consisted of the representativesof 8 taxonomic groups (Table 2) A total of 11 species ofMonogenea 1 species of Cestoda 3 species of Trematoda 2species of Nematoda 1 species of Hirudinea 1 species ofMollusca (larval stages termed glochidium) 1 species ofProtozoa and 2 species of Crustacea were identified (Table 2)The parasite infection level (expressed by prevalence abun-dance and intensity of infection) was compared between

BioMed Research International 3

22 Parasite Collection and Identification Monogeneanswere removed from the gills or fins placed into a drop ofwater on a slide coveredwith a coverslip andfixedusing glyc-erine ammonium picrate (GAP) for later analysis [32] Theremaining parasite species were fixed in 70 ethanol Parasitespecimens were examined using an Olympus model BX50light microscope (including an Olympus DP71 digital micro-scope camera) with phase contrast differential interferencecontrast and digital imaging (MicroImage 40 forWindows)

23 Ploidy Determination and Molecular Analyses A fin clipof about 1 cm2 in area was taken from each fish for ploidydetection and fixed in 70 ethanol Before analysis this tissuewas homogenised on a Petri dish in a 2ml solution of CyStainDNA 1 step Partec and relative DNA content was estimatedusing a Partec CCA I continuous flow cytometer (PartecGmbH httpswwwsysmex-parteccom) as was applied pre-viously for ploidymeasurements using blood sample [33 34]Fresh blood of diploid C auratus was obtained by punctureof the caudal blood vessel using a heparinised syringe andwas used as a reference standard Overall for our study 112diploid specimens (comprising 56 females and 56 males)and 91 triploid females were randomly selected The controlregion ofmitochondrial DNA (D-loop) was analysed for eachspecimen DNA was extracted by Dneasy Blood and TissueKit (Qiagen) from fin clips preserved in ethanol PCR andDNA sequencing were performed according to Sambrooket al [35] with small modifications following Vetesnık et al[36] and Papousek et al [37] For amplification the forwardprimer CarU32 (51015840-CCAAAGCCAGAATTCTAAAC-31015840) andthe reverse primer CarL509 (51015840-CATGTGGGGTAATGA-31015840)[37] were used A PCRproduct of 475ndash493 bp in length coversall ETAS domains (representing a high degree of variabilityconsidered useful for the study of intra- and interspeciesevolution) [38] and a part of the central domain in thecontrol region PCR was performed in a final volume of 30 120583lcontaining 1x PCR reaction buffer 15mMMgCl

2 03 120583M of

each primer (CarU32 and CarL509) 02mM dNTPs 15 UTaq polymerase and 100 ng120583l of template gDNA PCR wascarried out using the following steps 1min of denaturationat 95∘C followed by 32 cycles of 45 s at 94∘C 30 s at 50∘Cand 45 s at 72∘C and 10min of final elongation at 72∘C ThePCR products were checked on 15 agarose gel purified byHigh Pure PCR product purification kit (Roche) and directlysequenced using BigDye Terminator Cycle sequencing kit onan ABI 3130 Genetic Analyser (Applied Biosystems USA)The sequences were aligned using MEGA 60 [39] andanalysed using TCS 121 [40] The obtained sequences werecompared with the sequences for the mtDNA haplotypes ofthe C auratus complex available in GenBank The sequencesof mtDNA haplotypes obtained in this study were identicalwith the sequences available in GenBank under the followingaccession numbers FJ167410 FJ167411 FJ167413 FJ167414FJ167416 FJ167420 and FJ167423ndashFJ167425

24 Statistical Analyses General linear models (GLM) wereused to evaluate the threemain effects that is ploidy sex andyear on parasite abundance as well as two interaction effectsthat is ploidy with year and ploidy with body size The

standard body length of fish was included as a covariateParasite abundance was expressed by the following variablestotal parasite abundance (ie the abundance including allparasite specimens) total monogenean abundance (ie theabundance of the most common parasitic group) totalabundance of Nematoda (ie the abundance of the mostcommon endoparasite group) and the abundance of themost common parasitic species or genera that is Dactylo-gyrus spp (gill Monogenea) Gyrodactylus spp (gill and finMonogenea) and Ichthyophthirius multifiliis (Protozoa) Formultiple comparisons the Tukey post hoc test was used Allvariables applied in this study were examined for normaldistribution and homogeneity of variance For data that didnot fit these assumptions log-transformation was appliedStatistical analyses were performed in Statistica for Windows12 StatSoft Inc

3 Results

Random sampling of the mixed population of C gibelio inthe studied locality over a period of four years revealedthe following composition of the mixed gynogenetic-sexualpopulation 38 of fish were triploid specimens (37 weregynogenetic females and 1 were triploid males) and 63of fish were sexual diploid specimens Molecular analysis ofmtDNA confirmed that all specimens were representativesof the C auratus complex Our study revealed the presenceof 9 variants of 16 mtDNA haplotypes previously describedin the Southern Moravian region of the Czech Republic byPapousek [19] (Table 1) The Carassius gibelio lineage was themost frequent in the studied locality (96) Six mtDNAhaplotype variants of C gibelio were identified G01 G02G04 G05 G07 and G11 with G02 representing the mostcommon haplotype (Table 1) Two of the other haplotypesG01 and G04 were also common For sexual specimens G02and G04 were the most common haplotypes and G05 was araremitochondrial genotype in all four years of investigationFor gynogenetic females differences in haplotype frequenciesamong the four years were observed The most frequenthaplotypes of gynogenetic females reported in each yearwere as follows G01 G02 and G07 in 2012 G02 in 2013and G01 in both 2014 and 2015 Among the 6 C gibeliomtDNA haplotypes 3 haplotypes were found exclusively ingynogenetic females one was found exclusively in sexualspecimens and two were shared by sexual and gynogeneticspecimens The other three lineages C auratus (haplotypeA01) C langsdorfii (haplotype L01) and M line (haplotypeM01) were rare (24 05 and 1 resp) For the followinganalyses only the C gibelio lineage was selected due to thehigh frequency of this form

Parasite communities consisted of the representativesof 8 taxonomic groups (Table 2) A total of 11 species ofMonogenea 1 species of Cestoda 3 species of Trematoda 2species of Nematoda 1 species of Hirudinea 1 species ofMollusca (larval stages termed glochidium) 1 species ofProtozoa and 2 species of Crustacea were identified (Table 2)The parasite infection level (expressed by prevalence abun-dance and intensity of infection) was compared between

4 BioMed Research International

Table 1 Haplotypes of the mitochondrial control region (D-loop) in investigated specimens of Carassius auratus complex Total numbers ofspecimens and the numbers of specimens in each of four consecutive years (in parentheses) are shown for each mtDNA haplotype

C auratus complex Haplotype Number of diploids Number of triploids

C gibelio

G01 ndash 43 (1141414)G02 61 (1621159) 27 (7857)G04 34 (10969) 2 (0101)G05 9 (1251) ndashG07 ndash 11 (8201)G11 ndash 8 (4310)

C auratus A01 5 (3110) ndashC langsdorfii L01 1 (0010) ndashC auratusM line M01 2 (0101) ndash

gynogenetic and sexual forms The prevalence of Dactylo-gyrus spp and Gyrodactylus sprostonae (Monogenea) washigh in sexual males and females as well as in gynogeneticfemales (maximum prevalence was reported for 5 Dacty-logyrus species and a prevalence of more than 80 forGyrodactylus) More specifically Dactylogyrus dulkeiti Dintermedius D anchoratus D formosus and D inexpectatusand Gyrodactylus sprostonae represented the ectoparasitespecies reaching the highest intensities of infection in bothforms The prevalence of 5 other ectoparasite species thatis D vastator D baueri G shulmani G longoacuminatusand Ichthyophthirius multifiliis was moderate However theintensity of infection of I multifiliiswas also highThe preva-lence of other parasite species was low (le45) Concern-ing endoparasite species Schulmanela petruschewskii andPhilometroides sanguinea (Nematoda) achieved the highestintensity of infection especially in females (both gynogeneticand sexual) Total parasite abundance as well as monogeneanparasite abundance was lower in gynogenetic G07 haplotypeswhen compared to other haplotypes (MannndashWhitney test119901 = 0018 and 119901 = 0008 resp)

The effects of ploidy sex and year on parasite abundancewere analysed (Table 3) as described in Materials and Meth-ods Total parasite abundance (Figure 1(a)) as well as mono-genean abundance (Figure 1(b)) was significantly affected byyear body size and ploidy (through the interaction effectof ploidy and year) The highest levels of total parasiteabundance and monogenean abundance were reported in2015 the lowest in 2013 Sexual diploidsweremore parasitizedthan gynogenetic females in 2012 (119901 lt 005) and 2015 (119901 lt001) However no significant differences in total parasiteabundance or monogenean abundance were found between2013 and 2014

Dactylogyrus abundancewas significantly affected by yearand body size and there was also a weak but insignificanteffect of ploidy (119901 = 0127) through the interaction effectof ploidy and year (Figure 1(c) Table 3) Dactylogyrusabundance was significantly higher in 2015 compared to thefirst two years of investigation and was also higher in 2014compared to 2013 In 2015 Dactylogyrus abundance wasslightly higher in sexual diploids than in gynogenetic females(119901 = 0046) Gyrodactylus abundance (Figure 1(d)) was

significantly affected by year and ploidy through the inter-action effect of ploidy and year Gyrodactylus abundancereached its highest values in 2015 and was also higher in 2012compared to 2013 In 2012 and 2015 sexual diploids weremore parasitized by Gyrodactylus spp when compared togynogenetic females (119901 lt 001)

The abundance of I multifiliis was significantly affectedby year and body size and there was a small but insignificanteffect of ploidy (119901 = 014) through the interaction effect ofploidy and year (Table 3 Figure 1(e)) The lowest abundanceof I multifiliis was found in 2012 In 2012 and 2015 sexualdiploids were more parasitized by I multifiliis when com-pared to gynogenetic females

The abundance of Nematoda (two nematode species werepooled for the analysis) was significantly affected by yearploidy and body size (Table 3 Figure 1(f)) A significant inter-action effect of ploidy and body size onNematoda abundancewas also foundThe highest Nematoda abundance was foundin 2015 and the lowest in 2013 Overall females were moreparasitized than males (119901 = 0002) and gynogenetic femaleswere more parasitized than sexual diploids (119901 = 0010as evidenced in 2014 and 2015 see Figure 1(f)) Howeverwhen the effect of ploidy was analysed within each year asignificant difference in the abundance of Nematoda betweengynogenetic females and sexual diploids was found only in2015 (119901 = 0004)

4 Discussion

The Red Queen hypothesis describes host-parasite dynamicson the basis of negative parasite mediated frequency depen-dent selection which can represent one of the advantagesof sexual reproduction over asexual reproduction [2 2241 42] Thus we may expect that the asexual form of aspecies which exhibits low genetic variability will suffer fromhigher parasite load when compared to the sexual formwhich exhibits high genetic variability due to recombinationHowever as already mentioned there is no clear empiricalsupport for this hypothesis as studies focussing on parasiteload in coexisting asexual and sexual complexes do not showa consistent pattern In the case of the gynogenetic-sexual

BioMed Research International 5

Table2Prevalenceabu

ndance

(meanplusmnstandard

deviationSD

)andintensity

ofinfection(m

inim

umndashm

axim

um)for

parasitespeciesin

Carassius

gibelioPprevalenceAabu

ndanceand

Iintensity

ofinfection

Parasites

pecies

Diploid

males

Diploid

females

Triploid

females

P(

)A

IP(

)A

IP(

)A

IMean(plusmnSD

)Mean(plusmnSD

)Mean(plusmnSD

)

Mon

ogenea

Dactylogyrusd

ulkeiti

100

5529(6092)

6ndash353

100

5948(5110)

6ndash291

100

4868(3675)

12ndash187

Dactylogyrusintermedius

100

4502(4806)

8ndash301

100

4388(3844

)3ndash214

100

3930(3221)

5ndash196

Dactylogyrusa

nchoratus

100

6202(695

2)4ndash

419

100

6456(6044

)12ndash330

100

5877(5323)

5ndash372

Dactylogyrusformosus

100

1729

(1973)

1ndash118

981902(1861)

0ndash93

100

1668(1301)

1ndash90

Dactylogyrusinexpectatus

100

1788

(2028)

2ndash125

100

1581(1336)

2ndash76

100

1356(1020)

1ndash73

Dactylogyrusv

astator

77242

(345)

0ndash22

81275

(265)

0ndash15

6513

2(181)

0ndash10

Dactylogyrusb

aueri

4679

8(1382)

0ndash66

56523

(806)

0ndash46

63589

(850)

0ndash39

Gyrodactylu

ssprostona

e83

4281(13546

)0ndash

970

8710542

(26780)

0ndash1387

871885(2574)

0ndash180

Gyrodactylu

sshu

lmani

44279

(876

)0ndash

6242

256

(677)

0ndash35

23026

(053

)0ndash

3Gy

rodactylu

slongoacum

inatus

38260

(902)

0ndash62

58542

(1399)

0ndash71

60245

(363)

0ndash23

Paradiplozoonhomoion

8012

(047)

0ndash3

13017

(047)

0ndash2

7008

(030)

0ndash2

Cestoda

Caryoph

yllid

easp

2002

(014

)0-1

4073

(508)

0ndash37

ndashndash

ndash

Trem

atod

aAspidogaste

rsp

ndashndash

ndash8

015

(063)

0ndash4

5015

(073)

0ndash5

Diplosto

mum

sp

ndashndash

ndashndash

ndashndash

1001

(010

)0-1

Digenea

sp

15037

(124)

0ndash8

8023

(087)

0ndash5

7009

(035

)0ndash

2

Nem

atod

aPh

ilometroidessanguinea

23092

(294)

0ndash20

25240

(700)

0ndash46

4579

8(179

4)0ndash

103

Schu

lmanela

petru

schewskii

17346

(1340

)0ndash

7231

1027(4230)

0ndash299

22489

(297

1)0ndash

280

Hiru

dinea

Piscico

lageom

etra

2002

(014

)0-1

2002

(014

)0-1

4004

(020)

0-1

Mollusca

Glochidium

(Uniolarvalstages)

8008

(027)

0-1

6010

(045)

0ndash3

3005

(031

)0ndash

2Protozoa

Ichthyophthirius

multifi

liis

581152

(3258)

0ndash162

65656

(905)

0ndash36

4490

9(2695)

0ndash192

Crustacea

Ergasilus

sieboldi

15031

(089)

0ndash5

21062

(162)

0ndash8

13032

(097)

0ndash6

Argulusfoliaceus

13013

(034)

0-1

15017

(043)

0ndash2

18029

(094)

0ndash8

6 BioMed Research International

Table 3The effects of ploidy sex year and body size on parasite abundance (transformed in log) Parasite abundance was expressed by totalparasite abundance abundance of the most common parasitic groups genera or species in Carassius gibelio The statistically significant 119901values are shown in bold

Dependent variable Predicted variables 119865 119901 Total 119865 Total 119901

Total parasites

Year 14432 lt0001

10038 lt0001

Sex 0002 0960Ploidy 0002 0964

Body size 11572 0001Year lowast ploidy 3556 0015

Ploidy lowast body size 0001 0979

Monogenea

Year 13720 lt0001

9119 lt0001

Sex 0010 0922Ploidy 0206 0651

Body size 6660 0011Year lowast ploidy 4063 0008

Ploidy lowast body size 0279 0598

Dactylogyrus

Year 8387 lt0001

5530 lt0001

Sex 0482 0488Ploidy 0051 0822

Body size 6851 0010Year lowast ploidy 1927 0127

Ploidy lowast body size 0069 0794

Gyrodactylus

Year 18599 lt0001

9557 lt0001

Sex 0110 0740Ploidy 0038 0846

Body size 1446 0231Year lowast ploidy 4489 0005

Ploidy lowast body size 0073 0787

Ichthyophthirius multifiliis

Year 11887 lt0001

8268 lt0001

Sex 0210 0647Ploidy 0151 0698

Body size 8936 0003Year lowast ploidy 1847 0140

Ploidy lowast body size 0191 0662

Nematoda

Year 12201 lt0001

8217 lt0001

Sex 1884 0172Ploidy 7624 0006

Body size 14008 lt0001Year lowast ploidy 1191 0315

Ploidy lowast body size 7690 0006

complex including sexual Poeciliopsis monacha and the twocoexisting gynogenetic triploid clones of P 2monacha-lucidaLively et al [22] reported a higher accumulation of themetac-ercaria stage of the trematode Uvulifer sp causing black spotdisease in the most common triploid clone when comparedto sexual species and the less common gynogenetic clone ondifferent temporal and spatial scales (ie two different yearsof collection and three different pools) Infection by themetacercaria stage of the trematode Metagonimus sp alsoforming a black spot on the fish epidermis was analysed inthe gynogenetic-sexual complex of Carassius auratus langs-dorfiimdashC a burgeri byHakoyama et al [25] In this complexthe gynogenetic form is infrequent They suggested that thehigher parasite load in the gynogenetic formwas in part due

to the lower nonspecific immune reaction in the gynogeneticform compared to the sexual form In contrast no consistentpattern in parasite load between asexual Poecilia formosa andsexual P latipinna was observed on the spatial scale [43] Intheir study four populations were studiedmdashtwo exhibitingno difference in parasite load between asexual and sexualspecies one population exhibiting high overall parasite loadamong asexual species and the last exhibiting a greatermicro-parasite load among sexual species

In our study we revealed the same composition of para-site communities in both sexual and gynogenetic forms of Cgibelio that is a high number of parasite species with Mono-genea (ectoparasite group exhibiting generally high degree ofhost specificity) predominant and similar levels of prevalence

BioMed Research International 7

Tota

l par

asite

abun

danc

e

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(a)

Tota

l Mon

ogen

ea ab

unda

nce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(b)

Dac

tylo

gyru

sabu

ndan

ce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(c)

Gyro

dact

ylusa

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(d)

I m

ultifi

liis a

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(e)

Nem

atod

a abu

ndan

ce

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(f)

Figure 1 Parasite abundance (transformed in log) in sexual diploid form (F2n diploid females and M2n diploid males) and gynogenetictriploid females (F3n) Parasite abundance is expressed using the total parasites (a) Monogenea (b) Dactylogyrus species (c) Gyrodactylusspecies (d) Ichthyophthirius multifiliis (e) and Nematoda (f)

8 BioMed Research International

Surprisingly some monogenean species exhibited a trend ofhigher maximum intensity of infection in sexual specimenswhen compared to gynogenetic femalesThis trend was mostevident for Gyrodactylus sprostonae representing monoge-neans with a viviparous strategy (and very fast reproductionrate) and lower degree of host specificity when compared tooviparous and highly host specific gill monogeneans ofDactylogyrus On the other hand gynogenetic females tendedto be more parasitized with respect to the total numberof nematodes (but see below) However within-year com-parisons revealed higher monogenean abundance (for bothDactylogyrus and Gyrodactylus species) in the sexual formwhen compared to the gynogenetic form partially for 2012and particularly for 2015 when the overall level of para-site infection was the highest No obvious differences inmonogenean abundance were found in 2013 and in 2014Dactylogyrus abundance among gynogenetic females eventended to be slightly higher than Dactylogyrus abundanceamong the sexual form (though the difference was not signif-icant) Concerning endoparasites Nematoda were the mostabundant parasite group with two species reported in thisstudymdashPhilometroides sanguinea (parasite species restrictedto Carassius species) and Schulmanela petruschewskii (para-sitizing a wide range of fish species) The gynogenetic formwas more parasitized by nematodes than the sexual formHowever it seems that this result was generated by theeffect of body size as gynogenetic females reached greaterbody sizes than sexual specimens During all four years ofinvestigation gynogenetic females represented 30ndash40 ofthe mixed population sexual females 35ndash40 and sexualmales 22ndash30As a result the sexual formwasmore frequentin the habitat than the asexual form which may partiallyexplain the observed pattern of parasite infection Thecoexisting gynogenetic-sexual C gibelio complex was previ-ously investigated by Simkova et al [28] They found thatthere was a small difference in parasite load between sexualand gynogenetic forms (gynogenetic females suffered fromslightly higher parasite infection) However in their studythe selected mixed diploid-triploid population of C gibeliowas collected at two-time points (two consecutive years) withcoexisting gynogenetic females representing 51ndash54 of thepopulation sexual females representing 20ndash24 and sexualmales representing 20ndash22 A different ratio of sexual andasexual forms of C gibelio in a given population a differentpopulation density or the influence of unexamined abiotic orbiotic factors may potentially explain the difference betweenthe results observed in the present study and those obtainedpreviously by Simkova et al [28] as in the latter study thesampling of C gibelio was performed in a different localityIn addition in the study by Simkova et al [28] Gyrodactylusparasites represented only a small fraction of the observedparasite communities while Dactylogyrus parasites (with anoviparous strategy a slower reproduction rate and higherdegree of host specificity when compared to Gyrodactylus)predominated It is also possible that in some localities thecoexistence of gynogenetic and sexual forms of C gibelio isnot solely maintained by parasites and that some othermechanisms may be involved or even act more strongly

than parasites It is also possible that phylogenetically closelyrelated cyprinid species (ieCarassius carassius andCyprinuscarpio) living in sympatry with C gibelio act as suitable hostspecies for the transmission of host specific parasites in somelocalities However their occurrence was extremely rare inour system studiedThe general validity of the RQ hypothesiswith respect to how the coexistence of gynogenetic and sexualforms of C gibelio is maintained needs to be verified byexamining a wider range of mixed populations with differentratios of gynogenetic and sexual forms likely reflecting thespecific stages of evolution of this complex

Simkova et al [28] also performed genotyping usingthe major histocompatibility complex genes (MHC IIB) ofgynogenetic and sexual specimens They hypothesized andconfirmed that the most common gynogenetic clone is thetarget of parasite selection that is they showed that the mostfrequentMHCclones of gynogeneticC gibelio aremore para-sitized than sexual specimens or gynogenetic specimens withrare MHC genotypes In the present study no data concern-ing MHC genotype diversity was available Here we appliedan analysis ofmitochondrial haplotype diversity (based onD-loop sequence analysis) Concerning the mtDNA haplotypesin C gibelio in the Czech Republic previously studied byPapousek [19] G02 was the most common In the localitysampled for this study G02 was also the most common hap-lotype reported in the total sample of C gibelio and was morefrequently reported in sexual diploids than in gynogenetictriploid females In accordance with Papousek [19] weidentified G05 haplotype only in sexual diploids and G11 onlyin gynogenetic females However our data revealed G01 asthe most frequent haplotype in gynogenetic females (47 ofspecimens) In contrast to Papousek [19] we failed to identifythis haplotype in sexual diploid form Concerning overallhaplotype diversity the gynogenetic form exhibited a highernumber of haplotypes when compared to the sexual formWhen comparing haplotype frequencies among the fouryears similar frequencies of haplotypes were identified forthe sexual form while some changes in haplotype frequencywere observed for gynogenetic females However thesechanges were not commensurate with changes in parasiteinfection among years Parasite abundance was similar in allgynogenetic sexual and mixed haplotypes except for onerare gynogenetic haplotype namely G07 which was less par-asitized than other haplotypesThis less parasitized haplotypewas present mainly in fish sampled in 2012

5 Conclusions

In the present long-term study we investigated the composi-tions of parasite communities and levels of parasite infectionin coexisting gynogenetic and sexual forms of the cyprinidC gibelio representing the most common form of the Cauratus complex in the Czech Republic Gynogenetic andsexual forms shared almost all parasite species and overallexhibited similar levels of parasite infection However thelevel of parasite infection was variable for both forms overthe four years of investigation potentially as a result of sometemporal variability of multiple abiotic andor biotic factors

BioMed Research International 9

We found higher mtDNA haplotype diversity in the gyno-genetic form when compared to the sexual form Our resultsindicate that changes in the frequencies of mitochondrialgenotypes across years do not explain the observed temporalvariability in parasite infection Different patterns of parasiteinfection in sexual and gynogenetic forms of C gibelio werereported when comparisons were made within individualyears a finding which may potentially be associated withthe dynamics of host-parasite interactions However theobserved dynamics of parasite infection does not support theRQ hypothesis as we failed to find a significantly higher levelof parasite infection in the asexual form in any single year ofinvestigation With respect to future studies focused on thedynamics of host-parasite interactions we suggest the need toanalyse other genetic markers especially those related to hostimmunity

Conflicts of Interest

The authors declare no conflicts of interest regarding thepublication of this paper

Authorsrsquo Contributions

Andrea Simkova designed the study Tomas Pakosta analysedthe data Lukas Vetesnık collected fish material All authorswere involved in drafting and revising the manuscript Allauthors read and approved the final version of the manu-script

Acknowledgments

The study was funded by the Czech Science FoundationProject no P505120375 Andrea Simkova and TomasPakosta were partially funded by the Czech Science Founda-tion ECIP project no P50512G112 The authors are grate-ful to M Benovics L Gettova M L Cervenka Kicinja KKoukalova V Krasnovyd C Rahmouni and G Vagnerovafor their help with fish dissection and parasite collection andK Civanova and S Stierandova for their help with molecularanalyses They would like to thank K Halacka from theInstitute of Vertebrate Biology Academy of Sciences of theCzech Republic for his help with fish sampling They alsothank Matthew Nicholls for English language revision of themanuscript

References

[1] J M Smith The Evolution of Sex Cambridge University PressCambridge UK 1978

[2] G BellTheMasterpiece of Nature the Evolution and Genetics ofSexuality University of California Press Berkeley Calif USA1982

[3] D K Lamatsch andM Stock ldquoSperm-dependent parthenogen-esis and hybridogenesis in teleost fishesrdquo in Lost Sex I SchonK Martens and P Dijk Eds pp 399ndash432 Springer DordrechtNetherlands 2009

[4] F W Allendorf ldquoEcological and Genetic Effects of Fish Intro-ductions Synthesis and Recommendationsrdquo Canadian Journalof Fisheries andAquatic Sciences vol 48 no S1 pp 178ndash181 1991

[5] P B Moyle and T Light ldquoFish invasions in California Doabiotic factors determine successrdquo Ecology vol 77 no 6 pp1666ndash1670 1996

[6] I G Cowx ldquoIntroduction of fish species into european freshwaters Economic successes or ecological disasters rdquo BFPP -Bulletin Francais de la Peche et de la Protection des MilieuxAquatiques no 344-345 pp 57ndash77 1997

[7] S Lusk V Luskova and K Halacka ldquoPrussian carp ndash 25 yearssince its natural introductionrdquo in Proceedings of the 3rd CzechConference of Ichthyology pp 135ndash140 1998

[8] J A Szczerbowski P M Banarescu and H J Poepke ldquoTheFreshwater Fishes of Europe Cyprinidae 2 Part III Carassiusto Cyprinusrdquo AULA-Verlag vol 5 no 3 pp 5ndash41 2002

[9] S Lusk and V Luskova ldquoInvasive fish species in the CzechRepublicrdquo in Proceedings of the 8th Czech Conference of Ichthy-ology pp 135ndash140 Czech 2005

[10] E Garcıa-Berthou C Alcaraz Q Pou-Rovira L Zamora GCoenders and C Feo ldquoIntroduction pathways and establish-ment rates of invasive aquatic species in Europerdquo CanadianJournal of Fisheries and Aquatic Sciences vol 62 no 2 pp 453ndash463 2005

[11] S Lusk V Barus andV Vesely ldquoOn the occurrence ofCarassiusauratus in the Morava river drainage areardquo Folia Zoologica vol4 no 26 pp 377ndash381 1977

[12] S Lusk V Luskova and L Hanel ldquoAlien fish species in theCzech Republic and their impact on the native fish faunardquo FoliaZoologica vol 59 no 1 pp 57ndash72 2010

[13] S Lusk J Kosco V Luskova K Halacka and P Kosuth ldquoAlienfish species in the floodplains of theDyje and the Bodrog riversrdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 199ndash205 2004

[14] S Lusk V Barus and A Kirka ldquoCurrent spreading andimportance of the giebel (Carassius auratus gibelio Bloch) inCzechoslovakiardquo Czech Journal of Animal Science vol 25 no11 pp 871ndash878 1980 (Czech)

[15] V Barus and O Oliva Fauna of Czech Republic and SlovakRepublic Petromyzontes and Osteichtyes Academia PragueCzech Republic 1995

[16] V Luskova K Halacka L Vetesnık and S Lusk ldquoChanges ofploidy and sexuality status of lsquoCarassius auratusrsquo populationsin the drainage area of the River Dyje (Czech Republic)rdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 165ndash171 2004

[17] V Luskova S Lusk K Halacka and L Vetesnık ldquoCarassiusauratus gibeliomdashThe most successful invasive fish in waters ofthe Czech RepublicrdquoRussian Journal of Biological Invasions vol1 no 3 pp 176ndash180 2010

[18] L Vetesnık K Halacka V Luskova and S Lusk ldquoErythrocyteprofile of diploid and triploid silver crucian carp (Carassiusauratus)rdquo Acta Veterinaria Brno vol 75 no 2 pp 203ndash2072006

[19] I Papousek Molecular-genetic analyses of Carassius species incentral Europe [PhD thesis] Masaryk University Faculty ofScience Brno Czech Republic 2008

[20] L Van Valen ldquoA new evolutionary lawrdquo Evolutionary Theoryvol 1 pp 1ndash30 1973

[21] J Jaenike ldquoAn hypothesis to account for the maintenance ofsex within populationsrdquo EvolutionaryTheory vol 3 pp 191ndash1941978

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

International Journal of

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Anatomy Research International

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GenomicsInternational Journal of

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Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

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Hindawiwwwhindawicom Volume 2018

BioMed Research International

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Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

BioMed Research International 5

Table2Prevalenceabu

ndance

(meanplusmnstandard

deviationSD

)andintensity

ofinfection(m

inim

umndashm

axim

um)for

parasitespeciesin

Carassius

gibelioPprevalenceAabu

ndanceand

Iintensity

ofinfection

Parasites

pecies

Diploid

males

Diploid

females

Triploid

females

P(

)A

IP(

)A

IP(

)A

IMean(plusmnSD

)Mean(plusmnSD

)Mean(plusmnSD

)

Mon

ogenea

Dactylogyrusd

ulkeiti

100

5529(6092)

6ndash353

100

5948(5110)

6ndash291

100

4868(3675)

12ndash187

Dactylogyrusintermedius

100

4502(4806)

8ndash301

100

4388(3844

)3ndash214

100

3930(3221)

5ndash196

Dactylogyrusa

nchoratus

100

6202(695

2)4ndash

419

100

6456(6044

)12ndash330

100

5877(5323)

5ndash372

Dactylogyrusformosus

100

1729

(1973)

1ndash118

981902(1861)

0ndash93

100

1668(1301)

1ndash90

Dactylogyrusinexpectatus

100

1788

(2028)

2ndash125

100

1581(1336)

2ndash76

100

1356(1020)

1ndash73

Dactylogyrusv

astator

77242

(345)

0ndash22

81275

(265)

0ndash15

6513

2(181)

0ndash10

Dactylogyrusb

aueri

4679

8(1382)

0ndash66

56523

(806)

0ndash46

63589

(850)

0ndash39

Gyrodactylu

ssprostona

e83

4281(13546

)0ndash

970

8710542

(26780)

0ndash1387

871885(2574)

0ndash180

Gyrodactylu

sshu

lmani

44279

(876

)0ndash

6242

256

(677)

0ndash35

23026

(053

)0ndash

3Gy

rodactylu

slongoacum

inatus

38260

(902)

0ndash62

58542

(1399)

0ndash71

60245

(363)

0ndash23

Paradiplozoonhomoion

8012

(047)

0ndash3

13017

(047)

0ndash2

7008

(030)

0ndash2

Cestoda

Caryoph

yllid

easp

2002

(014

)0-1

4073

(508)

0ndash37

ndashndash

ndash

Trem

atod

aAspidogaste

rsp

ndashndash

ndash8

015

(063)

0ndash4

5015

(073)

0ndash5

Diplosto

mum

sp

ndashndash

ndashndash

ndashndash

1001

(010

)0-1

Digenea

sp

15037

(124)

0ndash8

8023

(087)

0ndash5

7009

(035

)0ndash

2

Nem

atod

aPh

ilometroidessanguinea

23092

(294)

0ndash20

25240

(700)

0ndash46

4579

8(179

4)0ndash

103

Schu

lmanela

petru

schewskii

17346

(1340

)0ndash

7231

1027(4230)

0ndash299

22489

(297

1)0ndash

280

Hiru

dinea

Piscico

lageom

etra

2002

(014

)0-1

2002

(014

)0-1

4004

(020)

0-1

Mollusca

Glochidium

(Uniolarvalstages)

8008

(027)

0-1

6010

(045)

0ndash3

3005

(031

)0ndash

2Protozoa

Ichthyophthirius

multifi

liis

581152

(3258)

0ndash162

65656

(905)

0ndash36

4490

9(2695)

0ndash192

Crustacea

Ergasilus

sieboldi

15031

(089)

0ndash5

21062

(162)

0ndash8

13032

(097)

0ndash6

Argulusfoliaceus

13013

(034)

0-1

15017

(043)

0ndash2

18029

(094)

0ndash8

6 BioMed Research International

Table 3The effects of ploidy sex year and body size on parasite abundance (transformed in log) Parasite abundance was expressed by totalparasite abundance abundance of the most common parasitic groups genera or species in Carassius gibelio The statistically significant 119901values are shown in bold

Dependent variable Predicted variables 119865 119901 Total 119865 Total 119901

Total parasites

Year 14432 lt0001

10038 lt0001

Sex 0002 0960Ploidy 0002 0964

Body size 11572 0001Year lowast ploidy 3556 0015

Ploidy lowast body size 0001 0979

Monogenea

Year 13720 lt0001

9119 lt0001

Sex 0010 0922Ploidy 0206 0651

Body size 6660 0011Year lowast ploidy 4063 0008

Ploidy lowast body size 0279 0598

Dactylogyrus

Year 8387 lt0001

5530 lt0001

Sex 0482 0488Ploidy 0051 0822

Body size 6851 0010Year lowast ploidy 1927 0127

Ploidy lowast body size 0069 0794

Gyrodactylus

Year 18599 lt0001

9557 lt0001

Sex 0110 0740Ploidy 0038 0846

Body size 1446 0231Year lowast ploidy 4489 0005

Ploidy lowast body size 0073 0787

Ichthyophthirius multifiliis

Year 11887 lt0001

8268 lt0001

Sex 0210 0647Ploidy 0151 0698

Body size 8936 0003Year lowast ploidy 1847 0140

Ploidy lowast body size 0191 0662

Nematoda

Year 12201 lt0001

8217 lt0001

Sex 1884 0172Ploidy 7624 0006

Body size 14008 lt0001Year lowast ploidy 1191 0315

Ploidy lowast body size 7690 0006

complex including sexual Poeciliopsis monacha and the twocoexisting gynogenetic triploid clones of P 2monacha-lucidaLively et al [22] reported a higher accumulation of themetac-ercaria stage of the trematode Uvulifer sp causing black spotdisease in the most common triploid clone when comparedto sexual species and the less common gynogenetic clone ondifferent temporal and spatial scales (ie two different yearsof collection and three different pools) Infection by themetacercaria stage of the trematode Metagonimus sp alsoforming a black spot on the fish epidermis was analysed inthe gynogenetic-sexual complex of Carassius auratus langs-dorfiimdashC a burgeri byHakoyama et al [25] In this complexthe gynogenetic form is infrequent They suggested that thehigher parasite load in the gynogenetic formwas in part due

to the lower nonspecific immune reaction in the gynogeneticform compared to the sexual form In contrast no consistentpattern in parasite load between asexual Poecilia formosa andsexual P latipinna was observed on the spatial scale [43] Intheir study four populations were studiedmdashtwo exhibitingno difference in parasite load between asexual and sexualspecies one population exhibiting high overall parasite loadamong asexual species and the last exhibiting a greatermicro-parasite load among sexual species

In our study we revealed the same composition of para-site communities in both sexual and gynogenetic forms of Cgibelio that is a high number of parasite species with Mono-genea (ectoparasite group exhibiting generally high degree ofhost specificity) predominant and similar levels of prevalence

BioMed Research International 7

Tota

l par

asite

abun

danc

e

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(a)

Tota

l Mon

ogen

ea ab

unda

nce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(b)

Dac

tylo

gyru

sabu

ndan

ce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(c)

Gyro

dact

ylusa

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(d)

I m

ultifi

liis a

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(e)

Nem

atod

a abu

ndan

ce

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(f)

Figure 1 Parasite abundance (transformed in log) in sexual diploid form (F2n diploid females and M2n diploid males) and gynogenetictriploid females (F3n) Parasite abundance is expressed using the total parasites (a) Monogenea (b) Dactylogyrus species (c) Gyrodactylusspecies (d) Ichthyophthirius multifiliis (e) and Nematoda (f)

8 BioMed Research International

Surprisingly some monogenean species exhibited a trend ofhigher maximum intensity of infection in sexual specimenswhen compared to gynogenetic femalesThis trend was mostevident for Gyrodactylus sprostonae representing monoge-neans with a viviparous strategy (and very fast reproductionrate) and lower degree of host specificity when compared tooviparous and highly host specific gill monogeneans ofDactylogyrus On the other hand gynogenetic females tendedto be more parasitized with respect to the total numberof nematodes (but see below) However within-year com-parisons revealed higher monogenean abundance (for bothDactylogyrus and Gyrodactylus species) in the sexual formwhen compared to the gynogenetic form partially for 2012and particularly for 2015 when the overall level of para-site infection was the highest No obvious differences inmonogenean abundance were found in 2013 and in 2014Dactylogyrus abundance among gynogenetic females eventended to be slightly higher than Dactylogyrus abundanceamong the sexual form (though the difference was not signif-icant) Concerning endoparasites Nematoda were the mostabundant parasite group with two species reported in thisstudymdashPhilometroides sanguinea (parasite species restrictedto Carassius species) and Schulmanela petruschewskii (para-sitizing a wide range of fish species) The gynogenetic formwas more parasitized by nematodes than the sexual formHowever it seems that this result was generated by theeffect of body size as gynogenetic females reached greaterbody sizes than sexual specimens During all four years ofinvestigation gynogenetic females represented 30ndash40 ofthe mixed population sexual females 35ndash40 and sexualmales 22ndash30As a result the sexual formwasmore frequentin the habitat than the asexual form which may partiallyexplain the observed pattern of parasite infection Thecoexisting gynogenetic-sexual C gibelio complex was previ-ously investigated by Simkova et al [28] They found thatthere was a small difference in parasite load between sexualand gynogenetic forms (gynogenetic females suffered fromslightly higher parasite infection) However in their studythe selected mixed diploid-triploid population of C gibeliowas collected at two-time points (two consecutive years) withcoexisting gynogenetic females representing 51ndash54 of thepopulation sexual females representing 20ndash24 and sexualmales representing 20ndash22 A different ratio of sexual andasexual forms of C gibelio in a given population a differentpopulation density or the influence of unexamined abiotic orbiotic factors may potentially explain the difference betweenthe results observed in the present study and those obtainedpreviously by Simkova et al [28] as in the latter study thesampling of C gibelio was performed in a different localityIn addition in the study by Simkova et al [28] Gyrodactylusparasites represented only a small fraction of the observedparasite communities while Dactylogyrus parasites (with anoviparous strategy a slower reproduction rate and higherdegree of host specificity when compared to Gyrodactylus)predominated It is also possible that in some localities thecoexistence of gynogenetic and sexual forms of C gibelio isnot solely maintained by parasites and that some othermechanisms may be involved or even act more strongly

than parasites It is also possible that phylogenetically closelyrelated cyprinid species (ieCarassius carassius andCyprinuscarpio) living in sympatry with C gibelio act as suitable hostspecies for the transmission of host specific parasites in somelocalities However their occurrence was extremely rare inour system studiedThe general validity of the RQ hypothesiswith respect to how the coexistence of gynogenetic and sexualforms of C gibelio is maintained needs to be verified byexamining a wider range of mixed populations with differentratios of gynogenetic and sexual forms likely reflecting thespecific stages of evolution of this complex

Simkova et al [28] also performed genotyping usingthe major histocompatibility complex genes (MHC IIB) ofgynogenetic and sexual specimens They hypothesized andconfirmed that the most common gynogenetic clone is thetarget of parasite selection that is they showed that the mostfrequentMHCclones of gynogeneticC gibelio aremore para-sitized than sexual specimens or gynogenetic specimens withrare MHC genotypes In the present study no data concern-ing MHC genotype diversity was available Here we appliedan analysis ofmitochondrial haplotype diversity (based onD-loop sequence analysis) Concerning the mtDNA haplotypesin C gibelio in the Czech Republic previously studied byPapousek [19] G02 was the most common In the localitysampled for this study G02 was also the most common hap-lotype reported in the total sample of C gibelio and was morefrequently reported in sexual diploids than in gynogenetictriploid females In accordance with Papousek [19] weidentified G05 haplotype only in sexual diploids and G11 onlyin gynogenetic females However our data revealed G01 asthe most frequent haplotype in gynogenetic females (47 ofspecimens) In contrast to Papousek [19] we failed to identifythis haplotype in sexual diploid form Concerning overallhaplotype diversity the gynogenetic form exhibited a highernumber of haplotypes when compared to the sexual formWhen comparing haplotype frequencies among the fouryears similar frequencies of haplotypes were identified forthe sexual form while some changes in haplotype frequencywere observed for gynogenetic females However thesechanges were not commensurate with changes in parasiteinfection among years Parasite abundance was similar in allgynogenetic sexual and mixed haplotypes except for onerare gynogenetic haplotype namely G07 which was less par-asitized than other haplotypesThis less parasitized haplotypewas present mainly in fish sampled in 2012

5 Conclusions

In the present long-term study we investigated the composi-tions of parasite communities and levels of parasite infectionin coexisting gynogenetic and sexual forms of the cyprinidC gibelio representing the most common form of the Cauratus complex in the Czech Republic Gynogenetic andsexual forms shared almost all parasite species and overallexhibited similar levels of parasite infection However thelevel of parasite infection was variable for both forms overthe four years of investigation potentially as a result of sometemporal variability of multiple abiotic andor biotic factors

BioMed Research International 9

We found higher mtDNA haplotype diversity in the gyno-genetic form when compared to the sexual form Our resultsindicate that changes in the frequencies of mitochondrialgenotypes across years do not explain the observed temporalvariability in parasite infection Different patterns of parasiteinfection in sexual and gynogenetic forms of C gibelio werereported when comparisons were made within individualyears a finding which may potentially be associated withthe dynamics of host-parasite interactions However theobserved dynamics of parasite infection does not support theRQ hypothesis as we failed to find a significantly higher levelof parasite infection in the asexual form in any single year ofinvestigation With respect to future studies focused on thedynamics of host-parasite interactions we suggest the need toanalyse other genetic markers especially those related to hostimmunity

Conflicts of Interest

The authors declare no conflicts of interest regarding thepublication of this paper

Authorsrsquo Contributions

Andrea Simkova designed the study Tomas Pakosta analysedthe data Lukas Vetesnık collected fish material All authorswere involved in drafting and revising the manuscript Allauthors read and approved the final version of the manu-script

Acknowledgments

The study was funded by the Czech Science FoundationProject no P505120375 Andrea Simkova and TomasPakosta were partially funded by the Czech Science Founda-tion ECIP project no P50512G112 The authors are grate-ful to M Benovics L Gettova M L Cervenka Kicinja KKoukalova V Krasnovyd C Rahmouni and G Vagnerovafor their help with fish dissection and parasite collection andK Civanova and S Stierandova for their help with molecularanalyses They would like to thank K Halacka from theInstitute of Vertebrate Biology Academy of Sciences of theCzech Republic for his help with fish sampling They alsothank Matthew Nicholls for English language revision of themanuscript

References

[1] J M Smith The Evolution of Sex Cambridge University PressCambridge UK 1978

[2] G BellTheMasterpiece of Nature the Evolution and Genetics ofSexuality University of California Press Berkeley Calif USA1982

[3] D K Lamatsch andM Stock ldquoSperm-dependent parthenogen-esis and hybridogenesis in teleost fishesrdquo in Lost Sex I SchonK Martens and P Dijk Eds pp 399ndash432 Springer DordrechtNetherlands 2009

[4] F W Allendorf ldquoEcological and Genetic Effects of Fish Intro-ductions Synthesis and Recommendationsrdquo Canadian Journalof Fisheries andAquatic Sciences vol 48 no S1 pp 178ndash181 1991

[5] P B Moyle and T Light ldquoFish invasions in California Doabiotic factors determine successrdquo Ecology vol 77 no 6 pp1666ndash1670 1996

[6] I G Cowx ldquoIntroduction of fish species into european freshwaters Economic successes or ecological disasters rdquo BFPP -Bulletin Francais de la Peche et de la Protection des MilieuxAquatiques no 344-345 pp 57ndash77 1997

[7] S Lusk V Luskova and K Halacka ldquoPrussian carp ndash 25 yearssince its natural introductionrdquo in Proceedings of the 3rd CzechConference of Ichthyology pp 135ndash140 1998

[8] J A Szczerbowski P M Banarescu and H J Poepke ldquoTheFreshwater Fishes of Europe Cyprinidae 2 Part III Carassiusto Cyprinusrdquo AULA-Verlag vol 5 no 3 pp 5ndash41 2002

[9] S Lusk and V Luskova ldquoInvasive fish species in the CzechRepublicrdquo in Proceedings of the 8th Czech Conference of Ichthy-ology pp 135ndash140 Czech 2005

[10] E Garcıa-Berthou C Alcaraz Q Pou-Rovira L Zamora GCoenders and C Feo ldquoIntroduction pathways and establish-ment rates of invasive aquatic species in Europerdquo CanadianJournal of Fisheries and Aquatic Sciences vol 62 no 2 pp 453ndash463 2005

[11] S Lusk V Barus andV Vesely ldquoOn the occurrence ofCarassiusauratus in the Morava river drainage areardquo Folia Zoologica vol4 no 26 pp 377ndash381 1977

[12] S Lusk V Luskova and L Hanel ldquoAlien fish species in theCzech Republic and their impact on the native fish faunardquo FoliaZoologica vol 59 no 1 pp 57ndash72 2010

[13] S Lusk J Kosco V Luskova K Halacka and P Kosuth ldquoAlienfish species in the floodplains of theDyje and the Bodrog riversrdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 199ndash205 2004

[14] S Lusk V Barus and A Kirka ldquoCurrent spreading andimportance of the giebel (Carassius auratus gibelio Bloch) inCzechoslovakiardquo Czech Journal of Animal Science vol 25 no11 pp 871ndash878 1980 (Czech)

[15] V Barus and O Oliva Fauna of Czech Republic and SlovakRepublic Petromyzontes and Osteichtyes Academia PragueCzech Republic 1995

[16] V Luskova K Halacka L Vetesnık and S Lusk ldquoChanges ofploidy and sexuality status of lsquoCarassius auratusrsquo populationsin the drainage area of the River Dyje (Czech Republic)rdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 165ndash171 2004

[17] V Luskova S Lusk K Halacka and L Vetesnık ldquoCarassiusauratus gibeliomdashThe most successful invasive fish in waters ofthe Czech RepublicrdquoRussian Journal of Biological Invasions vol1 no 3 pp 176ndash180 2010

[18] L Vetesnık K Halacka V Luskova and S Lusk ldquoErythrocyteprofile of diploid and triploid silver crucian carp (Carassiusauratus)rdquo Acta Veterinaria Brno vol 75 no 2 pp 203ndash2072006

[19] I Papousek Molecular-genetic analyses of Carassius species incentral Europe [PhD thesis] Masaryk University Faculty ofScience Brno Czech Republic 2008

[20] L Van Valen ldquoA new evolutionary lawrdquo Evolutionary Theoryvol 1 pp 1ndash30 1973

[21] J Jaenike ldquoAn hypothesis to account for the maintenance ofsex within populationsrdquo EvolutionaryTheory vol 3 pp 191ndash1941978

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

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Submit your manuscripts atwwwhindawicom

6 BioMed Research International

Table 3The effects of ploidy sex year and body size on parasite abundance (transformed in log) Parasite abundance was expressed by totalparasite abundance abundance of the most common parasitic groups genera or species in Carassius gibelio The statistically significant 119901values are shown in bold

Dependent variable Predicted variables 119865 119901 Total 119865 Total 119901

Total parasites

Year 14432 lt0001

10038 lt0001

Sex 0002 0960Ploidy 0002 0964

Body size 11572 0001Year lowast ploidy 3556 0015

Ploidy lowast body size 0001 0979

Monogenea

Year 13720 lt0001

9119 lt0001

Sex 0010 0922Ploidy 0206 0651

Body size 6660 0011Year lowast ploidy 4063 0008

Ploidy lowast body size 0279 0598

Dactylogyrus

Year 8387 lt0001

5530 lt0001

Sex 0482 0488Ploidy 0051 0822

Body size 6851 0010Year lowast ploidy 1927 0127

Ploidy lowast body size 0069 0794

Gyrodactylus

Year 18599 lt0001

9557 lt0001

Sex 0110 0740Ploidy 0038 0846

Body size 1446 0231Year lowast ploidy 4489 0005

Ploidy lowast body size 0073 0787

Ichthyophthirius multifiliis

Year 11887 lt0001

8268 lt0001

Sex 0210 0647Ploidy 0151 0698

Body size 8936 0003Year lowast ploidy 1847 0140

Ploidy lowast body size 0191 0662

Nematoda

Year 12201 lt0001

8217 lt0001

Sex 1884 0172Ploidy 7624 0006

Body size 14008 lt0001Year lowast ploidy 1191 0315

Ploidy lowast body size 7690 0006

complex including sexual Poeciliopsis monacha and the twocoexisting gynogenetic triploid clones of P 2monacha-lucidaLively et al [22] reported a higher accumulation of themetac-ercaria stage of the trematode Uvulifer sp causing black spotdisease in the most common triploid clone when comparedto sexual species and the less common gynogenetic clone ondifferent temporal and spatial scales (ie two different yearsof collection and three different pools) Infection by themetacercaria stage of the trematode Metagonimus sp alsoforming a black spot on the fish epidermis was analysed inthe gynogenetic-sexual complex of Carassius auratus langs-dorfiimdashC a burgeri byHakoyama et al [25] In this complexthe gynogenetic form is infrequent They suggested that thehigher parasite load in the gynogenetic formwas in part due

to the lower nonspecific immune reaction in the gynogeneticform compared to the sexual form In contrast no consistentpattern in parasite load between asexual Poecilia formosa andsexual P latipinna was observed on the spatial scale [43] Intheir study four populations were studiedmdashtwo exhibitingno difference in parasite load between asexual and sexualspecies one population exhibiting high overall parasite loadamong asexual species and the last exhibiting a greatermicro-parasite load among sexual species

In our study we revealed the same composition of para-site communities in both sexual and gynogenetic forms of Cgibelio that is a high number of parasite species with Mono-genea (ectoparasite group exhibiting generally high degree ofhost specificity) predominant and similar levels of prevalence

BioMed Research International 7

Tota

l par

asite

abun

danc

e

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(a)

Tota

l Mon

ogen

ea ab

unda

nce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(b)

Dac

tylo

gyru

sabu

ndan

ce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(c)

Gyro

dact

ylusa

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(d)

I m

ultifi

liis a

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(e)

Nem

atod

a abu

ndan

ce

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(f)

Figure 1 Parasite abundance (transformed in log) in sexual diploid form (F2n diploid females and M2n diploid males) and gynogenetictriploid females (F3n) Parasite abundance is expressed using the total parasites (a) Monogenea (b) Dactylogyrus species (c) Gyrodactylusspecies (d) Ichthyophthirius multifiliis (e) and Nematoda (f)

8 BioMed Research International

Surprisingly some monogenean species exhibited a trend ofhigher maximum intensity of infection in sexual specimenswhen compared to gynogenetic femalesThis trend was mostevident for Gyrodactylus sprostonae representing monoge-neans with a viviparous strategy (and very fast reproductionrate) and lower degree of host specificity when compared tooviparous and highly host specific gill monogeneans ofDactylogyrus On the other hand gynogenetic females tendedto be more parasitized with respect to the total numberof nematodes (but see below) However within-year com-parisons revealed higher monogenean abundance (for bothDactylogyrus and Gyrodactylus species) in the sexual formwhen compared to the gynogenetic form partially for 2012and particularly for 2015 when the overall level of para-site infection was the highest No obvious differences inmonogenean abundance were found in 2013 and in 2014Dactylogyrus abundance among gynogenetic females eventended to be slightly higher than Dactylogyrus abundanceamong the sexual form (though the difference was not signif-icant) Concerning endoparasites Nematoda were the mostabundant parasite group with two species reported in thisstudymdashPhilometroides sanguinea (parasite species restrictedto Carassius species) and Schulmanela petruschewskii (para-sitizing a wide range of fish species) The gynogenetic formwas more parasitized by nematodes than the sexual formHowever it seems that this result was generated by theeffect of body size as gynogenetic females reached greaterbody sizes than sexual specimens During all four years ofinvestigation gynogenetic females represented 30ndash40 ofthe mixed population sexual females 35ndash40 and sexualmales 22ndash30As a result the sexual formwasmore frequentin the habitat than the asexual form which may partiallyexplain the observed pattern of parasite infection Thecoexisting gynogenetic-sexual C gibelio complex was previ-ously investigated by Simkova et al [28] They found thatthere was a small difference in parasite load between sexualand gynogenetic forms (gynogenetic females suffered fromslightly higher parasite infection) However in their studythe selected mixed diploid-triploid population of C gibeliowas collected at two-time points (two consecutive years) withcoexisting gynogenetic females representing 51ndash54 of thepopulation sexual females representing 20ndash24 and sexualmales representing 20ndash22 A different ratio of sexual andasexual forms of C gibelio in a given population a differentpopulation density or the influence of unexamined abiotic orbiotic factors may potentially explain the difference betweenthe results observed in the present study and those obtainedpreviously by Simkova et al [28] as in the latter study thesampling of C gibelio was performed in a different localityIn addition in the study by Simkova et al [28] Gyrodactylusparasites represented only a small fraction of the observedparasite communities while Dactylogyrus parasites (with anoviparous strategy a slower reproduction rate and higherdegree of host specificity when compared to Gyrodactylus)predominated It is also possible that in some localities thecoexistence of gynogenetic and sexual forms of C gibelio isnot solely maintained by parasites and that some othermechanisms may be involved or even act more strongly

than parasites It is also possible that phylogenetically closelyrelated cyprinid species (ieCarassius carassius andCyprinuscarpio) living in sympatry with C gibelio act as suitable hostspecies for the transmission of host specific parasites in somelocalities However their occurrence was extremely rare inour system studiedThe general validity of the RQ hypothesiswith respect to how the coexistence of gynogenetic and sexualforms of C gibelio is maintained needs to be verified byexamining a wider range of mixed populations with differentratios of gynogenetic and sexual forms likely reflecting thespecific stages of evolution of this complex

Simkova et al [28] also performed genotyping usingthe major histocompatibility complex genes (MHC IIB) ofgynogenetic and sexual specimens They hypothesized andconfirmed that the most common gynogenetic clone is thetarget of parasite selection that is they showed that the mostfrequentMHCclones of gynogeneticC gibelio aremore para-sitized than sexual specimens or gynogenetic specimens withrare MHC genotypes In the present study no data concern-ing MHC genotype diversity was available Here we appliedan analysis ofmitochondrial haplotype diversity (based onD-loop sequence analysis) Concerning the mtDNA haplotypesin C gibelio in the Czech Republic previously studied byPapousek [19] G02 was the most common In the localitysampled for this study G02 was also the most common hap-lotype reported in the total sample of C gibelio and was morefrequently reported in sexual diploids than in gynogenetictriploid females In accordance with Papousek [19] weidentified G05 haplotype only in sexual diploids and G11 onlyin gynogenetic females However our data revealed G01 asthe most frequent haplotype in gynogenetic females (47 ofspecimens) In contrast to Papousek [19] we failed to identifythis haplotype in sexual diploid form Concerning overallhaplotype diversity the gynogenetic form exhibited a highernumber of haplotypes when compared to the sexual formWhen comparing haplotype frequencies among the fouryears similar frequencies of haplotypes were identified forthe sexual form while some changes in haplotype frequencywere observed for gynogenetic females However thesechanges were not commensurate with changes in parasiteinfection among years Parasite abundance was similar in allgynogenetic sexual and mixed haplotypes except for onerare gynogenetic haplotype namely G07 which was less par-asitized than other haplotypesThis less parasitized haplotypewas present mainly in fish sampled in 2012

5 Conclusions

In the present long-term study we investigated the composi-tions of parasite communities and levels of parasite infectionin coexisting gynogenetic and sexual forms of the cyprinidC gibelio representing the most common form of the Cauratus complex in the Czech Republic Gynogenetic andsexual forms shared almost all parasite species and overallexhibited similar levels of parasite infection However thelevel of parasite infection was variable for both forms overthe four years of investigation potentially as a result of sometemporal variability of multiple abiotic andor biotic factors

BioMed Research International 9

We found higher mtDNA haplotype diversity in the gyno-genetic form when compared to the sexual form Our resultsindicate that changes in the frequencies of mitochondrialgenotypes across years do not explain the observed temporalvariability in parasite infection Different patterns of parasiteinfection in sexual and gynogenetic forms of C gibelio werereported when comparisons were made within individualyears a finding which may potentially be associated withthe dynamics of host-parasite interactions However theobserved dynamics of parasite infection does not support theRQ hypothesis as we failed to find a significantly higher levelof parasite infection in the asexual form in any single year ofinvestigation With respect to future studies focused on thedynamics of host-parasite interactions we suggest the need toanalyse other genetic markers especially those related to hostimmunity

Conflicts of Interest

The authors declare no conflicts of interest regarding thepublication of this paper

Authorsrsquo Contributions

Andrea Simkova designed the study Tomas Pakosta analysedthe data Lukas Vetesnık collected fish material All authorswere involved in drafting and revising the manuscript Allauthors read and approved the final version of the manu-script

Acknowledgments

The study was funded by the Czech Science FoundationProject no P505120375 Andrea Simkova and TomasPakosta were partially funded by the Czech Science Founda-tion ECIP project no P50512G112 The authors are grate-ful to M Benovics L Gettova M L Cervenka Kicinja KKoukalova V Krasnovyd C Rahmouni and G Vagnerovafor their help with fish dissection and parasite collection andK Civanova and S Stierandova for their help with molecularanalyses They would like to thank K Halacka from theInstitute of Vertebrate Biology Academy of Sciences of theCzech Republic for his help with fish sampling They alsothank Matthew Nicholls for English language revision of themanuscript

References

[1] J M Smith The Evolution of Sex Cambridge University PressCambridge UK 1978

[2] G BellTheMasterpiece of Nature the Evolution and Genetics ofSexuality University of California Press Berkeley Calif USA1982

[3] D K Lamatsch andM Stock ldquoSperm-dependent parthenogen-esis and hybridogenesis in teleost fishesrdquo in Lost Sex I SchonK Martens and P Dijk Eds pp 399ndash432 Springer DordrechtNetherlands 2009

[4] F W Allendorf ldquoEcological and Genetic Effects of Fish Intro-ductions Synthesis and Recommendationsrdquo Canadian Journalof Fisheries andAquatic Sciences vol 48 no S1 pp 178ndash181 1991

[5] P B Moyle and T Light ldquoFish invasions in California Doabiotic factors determine successrdquo Ecology vol 77 no 6 pp1666ndash1670 1996

[6] I G Cowx ldquoIntroduction of fish species into european freshwaters Economic successes or ecological disasters rdquo BFPP -Bulletin Francais de la Peche et de la Protection des MilieuxAquatiques no 344-345 pp 57ndash77 1997

[7] S Lusk V Luskova and K Halacka ldquoPrussian carp ndash 25 yearssince its natural introductionrdquo in Proceedings of the 3rd CzechConference of Ichthyology pp 135ndash140 1998

[8] J A Szczerbowski P M Banarescu and H J Poepke ldquoTheFreshwater Fishes of Europe Cyprinidae 2 Part III Carassiusto Cyprinusrdquo AULA-Verlag vol 5 no 3 pp 5ndash41 2002

[9] S Lusk and V Luskova ldquoInvasive fish species in the CzechRepublicrdquo in Proceedings of the 8th Czech Conference of Ichthy-ology pp 135ndash140 Czech 2005

[10] E Garcıa-Berthou C Alcaraz Q Pou-Rovira L Zamora GCoenders and C Feo ldquoIntroduction pathways and establish-ment rates of invasive aquatic species in Europerdquo CanadianJournal of Fisheries and Aquatic Sciences vol 62 no 2 pp 453ndash463 2005

[11] S Lusk V Barus andV Vesely ldquoOn the occurrence ofCarassiusauratus in the Morava river drainage areardquo Folia Zoologica vol4 no 26 pp 377ndash381 1977

[12] S Lusk V Luskova and L Hanel ldquoAlien fish species in theCzech Republic and their impact on the native fish faunardquo FoliaZoologica vol 59 no 1 pp 57ndash72 2010

[13] S Lusk J Kosco V Luskova K Halacka and P Kosuth ldquoAlienfish species in the floodplains of theDyje and the Bodrog riversrdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 199ndash205 2004

[14] S Lusk V Barus and A Kirka ldquoCurrent spreading andimportance of the giebel (Carassius auratus gibelio Bloch) inCzechoslovakiardquo Czech Journal of Animal Science vol 25 no11 pp 871ndash878 1980 (Czech)

[15] V Barus and O Oliva Fauna of Czech Republic and SlovakRepublic Petromyzontes and Osteichtyes Academia PragueCzech Republic 1995

[16] V Luskova K Halacka L Vetesnık and S Lusk ldquoChanges ofploidy and sexuality status of lsquoCarassius auratusrsquo populationsin the drainage area of the River Dyje (Czech Republic)rdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 165ndash171 2004

[17] V Luskova S Lusk K Halacka and L Vetesnık ldquoCarassiusauratus gibeliomdashThe most successful invasive fish in waters ofthe Czech RepublicrdquoRussian Journal of Biological Invasions vol1 no 3 pp 176ndash180 2010

[18] L Vetesnık K Halacka V Luskova and S Lusk ldquoErythrocyteprofile of diploid and triploid silver crucian carp (Carassiusauratus)rdquo Acta Veterinaria Brno vol 75 no 2 pp 203ndash2072006

[19] I Papousek Molecular-genetic analyses of Carassius species incentral Europe [PhD thesis] Masaryk University Faculty ofScience Brno Czech Republic 2008

[20] L Van Valen ldquoA new evolutionary lawrdquo Evolutionary Theoryvol 1 pp 1ndash30 1973

[21] J Jaenike ldquoAn hypothesis to account for the maintenance ofsex within populationsrdquo EvolutionaryTheory vol 3 pp 191ndash1941978

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

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Hindawiwwwhindawicom Volume 2018

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GenomicsInternational Journal of

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Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

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Hindawiwwwhindawicom Volume 2018

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Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

BioMed Research International 7

Tota

l par

asite

abun

danc

e

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(a)

Tota

l Mon

ogen

ea ab

unda

nce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(b)

Dac

tylo

gyru

sabu

ndan

ce

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n30

28

26

24

22

20

18

Mean－Ｈ plusmn ３

(c)

Gyro

dact

ylusa

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(d)

I m

ultifi

liis a

bund

ance

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(e)

Nem

atod

a abu

ndan

ce

24

20

16

12

08

04

00

2012

_F 2

n2012

_F3

n2012

_M2

n2013

_F 2

n2013

_F 3

n2013

_M 2

n2014

_F 2

n2014

_F 3

n2014

_M 2

n2015

_F 2

n2015

_F 3

n2015

_M 2

n

Mean－Ｈ plusmn ３

(f)

Figure 1 Parasite abundance (transformed in log) in sexual diploid form (F2n diploid females and M2n diploid males) and gynogenetictriploid females (F3n) Parasite abundance is expressed using the total parasites (a) Monogenea (b) Dactylogyrus species (c) Gyrodactylusspecies (d) Ichthyophthirius multifiliis (e) and Nematoda (f)

8 BioMed Research International

Surprisingly some monogenean species exhibited a trend ofhigher maximum intensity of infection in sexual specimenswhen compared to gynogenetic femalesThis trend was mostevident for Gyrodactylus sprostonae representing monoge-neans with a viviparous strategy (and very fast reproductionrate) and lower degree of host specificity when compared tooviparous and highly host specific gill monogeneans ofDactylogyrus On the other hand gynogenetic females tendedto be more parasitized with respect to the total numberof nematodes (but see below) However within-year com-parisons revealed higher monogenean abundance (for bothDactylogyrus and Gyrodactylus species) in the sexual formwhen compared to the gynogenetic form partially for 2012and particularly for 2015 when the overall level of para-site infection was the highest No obvious differences inmonogenean abundance were found in 2013 and in 2014Dactylogyrus abundance among gynogenetic females eventended to be slightly higher than Dactylogyrus abundanceamong the sexual form (though the difference was not signif-icant) Concerning endoparasites Nematoda were the mostabundant parasite group with two species reported in thisstudymdashPhilometroides sanguinea (parasite species restrictedto Carassius species) and Schulmanela petruschewskii (para-sitizing a wide range of fish species) The gynogenetic formwas more parasitized by nematodes than the sexual formHowever it seems that this result was generated by theeffect of body size as gynogenetic females reached greaterbody sizes than sexual specimens During all four years ofinvestigation gynogenetic females represented 30ndash40 ofthe mixed population sexual females 35ndash40 and sexualmales 22ndash30As a result the sexual formwasmore frequentin the habitat than the asexual form which may partiallyexplain the observed pattern of parasite infection Thecoexisting gynogenetic-sexual C gibelio complex was previ-ously investigated by Simkova et al [28] They found thatthere was a small difference in parasite load between sexualand gynogenetic forms (gynogenetic females suffered fromslightly higher parasite infection) However in their studythe selected mixed diploid-triploid population of C gibeliowas collected at two-time points (two consecutive years) withcoexisting gynogenetic females representing 51ndash54 of thepopulation sexual females representing 20ndash24 and sexualmales representing 20ndash22 A different ratio of sexual andasexual forms of C gibelio in a given population a differentpopulation density or the influence of unexamined abiotic orbiotic factors may potentially explain the difference betweenthe results observed in the present study and those obtainedpreviously by Simkova et al [28] as in the latter study thesampling of C gibelio was performed in a different localityIn addition in the study by Simkova et al [28] Gyrodactylusparasites represented only a small fraction of the observedparasite communities while Dactylogyrus parasites (with anoviparous strategy a slower reproduction rate and higherdegree of host specificity when compared to Gyrodactylus)predominated It is also possible that in some localities thecoexistence of gynogenetic and sexual forms of C gibelio isnot solely maintained by parasites and that some othermechanisms may be involved or even act more strongly

than parasites It is also possible that phylogenetically closelyrelated cyprinid species (ieCarassius carassius andCyprinuscarpio) living in sympatry with C gibelio act as suitable hostspecies for the transmission of host specific parasites in somelocalities However their occurrence was extremely rare inour system studiedThe general validity of the RQ hypothesiswith respect to how the coexistence of gynogenetic and sexualforms of C gibelio is maintained needs to be verified byexamining a wider range of mixed populations with differentratios of gynogenetic and sexual forms likely reflecting thespecific stages of evolution of this complex

Simkova et al [28] also performed genotyping usingthe major histocompatibility complex genes (MHC IIB) ofgynogenetic and sexual specimens They hypothesized andconfirmed that the most common gynogenetic clone is thetarget of parasite selection that is they showed that the mostfrequentMHCclones of gynogeneticC gibelio aremore para-sitized than sexual specimens or gynogenetic specimens withrare MHC genotypes In the present study no data concern-ing MHC genotype diversity was available Here we appliedan analysis ofmitochondrial haplotype diversity (based onD-loop sequence analysis) Concerning the mtDNA haplotypesin C gibelio in the Czech Republic previously studied byPapousek [19] G02 was the most common In the localitysampled for this study G02 was also the most common hap-lotype reported in the total sample of C gibelio and was morefrequently reported in sexual diploids than in gynogenetictriploid females In accordance with Papousek [19] weidentified G05 haplotype only in sexual diploids and G11 onlyin gynogenetic females However our data revealed G01 asthe most frequent haplotype in gynogenetic females (47 ofspecimens) In contrast to Papousek [19] we failed to identifythis haplotype in sexual diploid form Concerning overallhaplotype diversity the gynogenetic form exhibited a highernumber of haplotypes when compared to the sexual formWhen comparing haplotype frequencies among the fouryears similar frequencies of haplotypes were identified forthe sexual form while some changes in haplotype frequencywere observed for gynogenetic females However thesechanges were not commensurate with changes in parasiteinfection among years Parasite abundance was similar in allgynogenetic sexual and mixed haplotypes except for onerare gynogenetic haplotype namely G07 which was less par-asitized than other haplotypesThis less parasitized haplotypewas present mainly in fish sampled in 2012

5 Conclusions

In the present long-term study we investigated the composi-tions of parasite communities and levels of parasite infectionin coexisting gynogenetic and sexual forms of the cyprinidC gibelio representing the most common form of the Cauratus complex in the Czech Republic Gynogenetic andsexual forms shared almost all parasite species and overallexhibited similar levels of parasite infection However thelevel of parasite infection was variable for both forms overthe four years of investigation potentially as a result of sometemporal variability of multiple abiotic andor biotic factors

BioMed Research International 9

We found higher mtDNA haplotype diversity in the gyno-genetic form when compared to the sexual form Our resultsindicate that changes in the frequencies of mitochondrialgenotypes across years do not explain the observed temporalvariability in parasite infection Different patterns of parasiteinfection in sexual and gynogenetic forms of C gibelio werereported when comparisons were made within individualyears a finding which may potentially be associated withthe dynamics of host-parasite interactions However theobserved dynamics of parasite infection does not support theRQ hypothesis as we failed to find a significantly higher levelof parasite infection in the asexual form in any single year ofinvestigation With respect to future studies focused on thedynamics of host-parasite interactions we suggest the need toanalyse other genetic markers especially those related to hostimmunity

Conflicts of Interest

The authors declare no conflicts of interest regarding thepublication of this paper

Authorsrsquo Contributions

Andrea Simkova designed the study Tomas Pakosta analysedthe data Lukas Vetesnık collected fish material All authorswere involved in drafting and revising the manuscript Allauthors read and approved the final version of the manu-script

Acknowledgments

The study was funded by the Czech Science FoundationProject no P505120375 Andrea Simkova and TomasPakosta were partially funded by the Czech Science Founda-tion ECIP project no P50512G112 The authors are grate-ful to M Benovics L Gettova M L Cervenka Kicinja KKoukalova V Krasnovyd C Rahmouni and G Vagnerovafor their help with fish dissection and parasite collection andK Civanova and S Stierandova for their help with molecularanalyses They would like to thank K Halacka from theInstitute of Vertebrate Biology Academy of Sciences of theCzech Republic for his help with fish sampling They alsothank Matthew Nicholls for English language revision of themanuscript

References

[1] J M Smith The Evolution of Sex Cambridge University PressCambridge UK 1978

[2] G BellTheMasterpiece of Nature the Evolution and Genetics ofSexuality University of California Press Berkeley Calif USA1982

[3] D K Lamatsch andM Stock ldquoSperm-dependent parthenogen-esis and hybridogenesis in teleost fishesrdquo in Lost Sex I SchonK Martens and P Dijk Eds pp 399ndash432 Springer DordrechtNetherlands 2009

[4] F W Allendorf ldquoEcological and Genetic Effects of Fish Intro-ductions Synthesis and Recommendationsrdquo Canadian Journalof Fisheries andAquatic Sciences vol 48 no S1 pp 178ndash181 1991

[5] P B Moyle and T Light ldquoFish invasions in California Doabiotic factors determine successrdquo Ecology vol 77 no 6 pp1666ndash1670 1996

[6] I G Cowx ldquoIntroduction of fish species into european freshwaters Economic successes or ecological disasters rdquo BFPP -Bulletin Francais de la Peche et de la Protection des MilieuxAquatiques no 344-345 pp 57ndash77 1997

[7] S Lusk V Luskova and K Halacka ldquoPrussian carp ndash 25 yearssince its natural introductionrdquo in Proceedings of the 3rd CzechConference of Ichthyology pp 135ndash140 1998

[8] J A Szczerbowski P M Banarescu and H J Poepke ldquoTheFreshwater Fishes of Europe Cyprinidae 2 Part III Carassiusto Cyprinusrdquo AULA-Verlag vol 5 no 3 pp 5ndash41 2002

[9] S Lusk and V Luskova ldquoInvasive fish species in the CzechRepublicrdquo in Proceedings of the 8th Czech Conference of Ichthy-ology pp 135ndash140 Czech 2005

[10] E Garcıa-Berthou C Alcaraz Q Pou-Rovira L Zamora GCoenders and C Feo ldquoIntroduction pathways and establish-ment rates of invasive aquatic species in Europerdquo CanadianJournal of Fisheries and Aquatic Sciences vol 62 no 2 pp 453ndash463 2005

[11] S Lusk V Barus andV Vesely ldquoOn the occurrence ofCarassiusauratus in the Morava river drainage areardquo Folia Zoologica vol4 no 26 pp 377ndash381 1977

[12] S Lusk V Luskova and L Hanel ldquoAlien fish species in theCzech Republic and their impact on the native fish faunardquo FoliaZoologica vol 59 no 1 pp 57ndash72 2010

[13] S Lusk J Kosco V Luskova K Halacka and P Kosuth ldquoAlienfish species in the floodplains of theDyje and the Bodrog riversrdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 199ndash205 2004

[14] S Lusk V Barus and A Kirka ldquoCurrent spreading andimportance of the giebel (Carassius auratus gibelio Bloch) inCzechoslovakiardquo Czech Journal of Animal Science vol 25 no11 pp 871ndash878 1980 (Czech)

[15] V Barus and O Oliva Fauna of Czech Republic and SlovakRepublic Petromyzontes and Osteichtyes Academia PragueCzech Republic 1995

[16] V Luskova K Halacka L Vetesnık and S Lusk ldquoChanges ofploidy and sexuality status of lsquoCarassius auratusrsquo populationsin the drainage area of the River Dyje (Czech Republic)rdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 165ndash171 2004

[17] V Luskova S Lusk K Halacka and L Vetesnık ldquoCarassiusauratus gibeliomdashThe most successful invasive fish in waters ofthe Czech RepublicrdquoRussian Journal of Biological Invasions vol1 no 3 pp 176ndash180 2010

[18] L Vetesnık K Halacka V Luskova and S Lusk ldquoErythrocyteprofile of diploid and triploid silver crucian carp (Carassiusauratus)rdquo Acta Veterinaria Brno vol 75 no 2 pp 203ndash2072006

[19] I Papousek Molecular-genetic analyses of Carassius species incentral Europe [PhD thesis] Masaryk University Faculty ofScience Brno Czech Republic 2008

[20] L Van Valen ldquoA new evolutionary lawrdquo Evolutionary Theoryvol 1 pp 1ndash30 1973

[21] J Jaenike ldquoAn hypothesis to account for the maintenance ofsex within populationsrdquo EvolutionaryTheory vol 3 pp 191ndash1941978

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

8 BioMed Research International

Surprisingly some monogenean species exhibited a trend ofhigher maximum intensity of infection in sexual specimenswhen compared to gynogenetic femalesThis trend was mostevident for Gyrodactylus sprostonae representing monoge-neans with a viviparous strategy (and very fast reproductionrate) and lower degree of host specificity when compared tooviparous and highly host specific gill monogeneans ofDactylogyrus On the other hand gynogenetic females tendedto be more parasitized with respect to the total numberof nematodes (but see below) However within-year com-parisons revealed higher monogenean abundance (for bothDactylogyrus and Gyrodactylus species) in the sexual formwhen compared to the gynogenetic form partially for 2012and particularly for 2015 when the overall level of para-site infection was the highest No obvious differences inmonogenean abundance were found in 2013 and in 2014Dactylogyrus abundance among gynogenetic females eventended to be slightly higher than Dactylogyrus abundanceamong the sexual form (though the difference was not signif-icant) Concerning endoparasites Nematoda were the mostabundant parasite group with two species reported in thisstudymdashPhilometroides sanguinea (parasite species restrictedto Carassius species) and Schulmanela petruschewskii (para-sitizing a wide range of fish species) The gynogenetic formwas more parasitized by nematodes than the sexual formHowever it seems that this result was generated by theeffect of body size as gynogenetic females reached greaterbody sizes than sexual specimens During all four years ofinvestigation gynogenetic females represented 30ndash40 ofthe mixed population sexual females 35ndash40 and sexualmales 22ndash30As a result the sexual formwasmore frequentin the habitat than the asexual form which may partiallyexplain the observed pattern of parasite infection Thecoexisting gynogenetic-sexual C gibelio complex was previ-ously investigated by Simkova et al [28] They found thatthere was a small difference in parasite load between sexualand gynogenetic forms (gynogenetic females suffered fromslightly higher parasite infection) However in their studythe selected mixed diploid-triploid population of C gibeliowas collected at two-time points (two consecutive years) withcoexisting gynogenetic females representing 51ndash54 of thepopulation sexual females representing 20ndash24 and sexualmales representing 20ndash22 A different ratio of sexual andasexual forms of C gibelio in a given population a differentpopulation density or the influence of unexamined abiotic orbiotic factors may potentially explain the difference betweenthe results observed in the present study and those obtainedpreviously by Simkova et al [28] as in the latter study thesampling of C gibelio was performed in a different localityIn addition in the study by Simkova et al [28] Gyrodactylusparasites represented only a small fraction of the observedparasite communities while Dactylogyrus parasites (with anoviparous strategy a slower reproduction rate and higherdegree of host specificity when compared to Gyrodactylus)predominated It is also possible that in some localities thecoexistence of gynogenetic and sexual forms of C gibelio isnot solely maintained by parasites and that some othermechanisms may be involved or even act more strongly

than parasites It is also possible that phylogenetically closelyrelated cyprinid species (ieCarassius carassius andCyprinuscarpio) living in sympatry with C gibelio act as suitable hostspecies for the transmission of host specific parasites in somelocalities However their occurrence was extremely rare inour system studiedThe general validity of the RQ hypothesiswith respect to how the coexistence of gynogenetic and sexualforms of C gibelio is maintained needs to be verified byexamining a wider range of mixed populations with differentratios of gynogenetic and sexual forms likely reflecting thespecific stages of evolution of this complex

Simkova et al [28] also performed genotyping usingthe major histocompatibility complex genes (MHC IIB) ofgynogenetic and sexual specimens They hypothesized andconfirmed that the most common gynogenetic clone is thetarget of parasite selection that is they showed that the mostfrequentMHCclones of gynogeneticC gibelio aremore para-sitized than sexual specimens or gynogenetic specimens withrare MHC genotypes In the present study no data concern-ing MHC genotype diversity was available Here we appliedan analysis ofmitochondrial haplotype diversity (based onD-loop sequence analysis) Concerning the mtDNA haplotypesin C gibelio in the Czech Republic previously studied byPapousek [19] G02 was the most common In the localitysampled for this study G02 was also the most common hap-lotype reported in the total sample of C gibelio and was morefrequently reported in sexual diploids than in gynogenetictriploid females In accordance with Papousek [19] weidentified G05 haplotype only in sexual diploids and G11 onlyin gynogenetic females However our data revealed G01 asthe most frequent haplotype in gynogenetic females (47 ofspecimens) In contrast to Papousek [19] we failed to identifythis haplotype in sexual diploid form Concerning overallhaplotype diversity the gynogenetic form exhibited a highernumber of haplotypes when compared to the sexual formWhen comparing haplotype frequencies among the fouryears similar frequencies of haplotypes were identified forthe sexual form while some changes in haplotype frequencywere observed for gynogenetic females However thesechanges were not commensurate with changes in parasiteinfection among years Parasite abundance was similar in allgynogenetic sexual and mixed haplotypes except for onerare gynogenetic haplotype namely G07 which was less par-asitized than other haplotypesThis less parasitized haplotypewas present mainly in fish sampled in 2012

5 Conclusions

In the present long-term study we investigated the composi-tions of parasite communities and levels of parasite infectionin coexisting gynogenetic and sexual forms of the cyprinidC gibelio representing the most common form of the Cauratus complex in the Czech Republic Gynogenetic andsexual forms shared almost all parasite species and overallexhibited similar levels of parasite infection However thelevel of parasite infection was variable for both forms overthe four years of investigation potentially as a result of sometemporal variability of multiple abiotic andor biotic factors

BioMed Research International 9

We found higher mtDNA haplotype diversity in the gyno-genetic form when compared to the sexual form Our resultsindicate that changes in the frequencies of mitochondrialgenotypes across years do not explain the observed temporalvariability in parasite infection Different patterns of parasiteinfection in sexual and gynogenetic forms of C gibelio werereported when comparisons were made within individualyears a finding which may potentially be associated withthe dynamics of host-parasite interactions However theobserved dynamics of parasite infection does not support theRQ hypothesis as we failed to find a significantly higher levelof parasite infection in the asexual form in any single year ofinvestigation With respect to future studies focused on thedynamics of host-parasite interactions we suggest the need toanalyse other genetic markers especially those related to hostimmunity

Conflicts of Interest

The authors declare no conflicts of interest regarding thepublication of this paper

Authorsrsquo Contributions

Andrea Simkova designed the study Tomas Pakosta analysedthe data Lukas Vetesnık collected fish material All authorswere involved in drafting and revising the manuscript Allauthors read and approved the final version of the manu-script

Acknowledgments

The study was funded by the Czech Science FoundationProject no P505120375 Andrea Simkova and TomasPakosta were partially funded by the Czech Science Founda-tion ECIP project no P50512G112 The authors are grate-ful to M Benovics L Gettova M L Cervenka Kicinja KKoukalova V Krasnovyd C Rahmouni and G Vagnerovafor their help with fish dissection and parasite collection andK Civanova and S Stierandova for their help with molecularanalyses They would like to thank K Halacka from theInstitute of Vertebrate Biology Academy of Sciences of theCzech Republic for his help with fish sampling They alsothank Matthew Nicholls for English language revision of themanuscript

References

[1] J M Smith The Evolution of Sex Cambridge University PressCambridge UK 1978

[2] G BellTheMasterpiece of Nature the Evolution and Genetics ofSexuality University of California Press Berkeley Calif USA1982

[3] D K Lamatsch andM Stock ldquoSperm-dependent parthenogen-esis and hybridogenesis in teleost fishesrdquo in Lost Sex I SchonK Martens and P Dijk Eds pp 399ndash432 Springer DordrechtNetherlands 2009

[4] F W Allendorf ldquoEcological and Genetic Effects of Fish Intro-ductions Synthesis and Recommendationsrdquo Canadian Journalof Fisheries andAquatic Sciences vol 48 no S1 pp 178ndash181 1991

[5] P B Moyle and T Light ldquoFish invasions in California Doabiotic factors determine successrdquo Ecology vol 77 no 6 pp1666ndash1670 1996

[6] I G Cowx ldquoIntroduction of fish species into european freshwaters Economic successes or ecological disasters rdquo BFPP -Bulletin Francais de la Peche et de la Protection des MilieuxAquatiques no 344-345 pp 57ndash77 1997

[7] S Lusk V Luskova and K Halacka ldquoPrussian carp ndash 25 yearssince its natural introductionrdquo in Proceedings of the 3rd CzechConference of Ichthyology pp 135ndash140 1998

[8] J A Szczerbowski P M Banarescu and H J Poepke ldquoTheFreshwater Fishes of Europe Cyprinidae 2 Part III Carassiusto Cyprinusrdquo AULA-Verlag vol 5 no 3 pp 5ndash41 2002

[9] S Lusk and V Luskova ldquoInvasive fish species in the CzechRepublicrdquo in Proceedings of the 8th Czech Conference of Ichthy-ology pp 135ndash140 Czech 2005

[10] E Garcıa-Berthou C Alcaraz Q Pou-Rovira L Zamora GCoenders and C Feo ldquoIntroduction pathways and establish-ment rates of invasive aquatic species in Europerdquo CanadianJournal of Fisheries and Aquatic Sciences vol 62 no 2 pp 453ndash463 2005

[11] S Lusk V Barus andV Vesely ldquoOn the occurrence ofCarassiusauratus in the Morava river drainage areardquo Folia Zoologica vol4 no 26 pp 377ndash381 1977

[12] S Lusk V Luskova and L Hanel ldquoAlien fish species in theCzech Republic and their impact on the native fish faunardquo FoliaZoologica vol 59 no 1 pp 57ndash72 2010

[13] S Lusk J Kosco V Luskova K Halacka and P Kosuth ldquoAlienfish species in the floodplains of theDyje and the Bodrog riversrdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 199ndash205 2004

[14] S Lusk V Barus and A Kirka ldquoCurrent spreading andimportance of the giebel (Carassius auratus gibelio Bloch) inCzechoslovakiardquo Czech Journal of Animal Science vol 25 no11 pp 871ndash878 1980 (Czech)

[15] V Barus and O Oliva Fauna of Czech Republic and SlovakRepublic Petromyzontes and Osteichtyes Academia PragueCzech Republic 1995

[16] V Luskova K Halacka L Vetesnık and S Lusk ldquoChanges ofploidy and sexuality status of lsquoCarassius auratusrsquo populationsin the drainage area of the River Dyje (Czech Republic)rdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 165ndash171 2004

[17] V Luskova S Lusk K Halacka and L Vetesnık ldquoCarassiusauratus gibeliomdashThe most successful invasive fish in waters ofthe Czech RepublicrdquoRussian Journal of Biological Invasions vol1 no 3 pp 176ndash180 2010

[18] L Vetesnık K Halacka V Luskova and S Lusk ldquoErythrocyteprofile of diploid and triploid silver crucian carp (Carassiusauratus)rdquo Acta Veterinaria Brno vol 75 no 2 pp 203ndash2072006

[19] I Papousek Molecular-genetic analyses of Carassius species incentral Europe [PhD thesis] Masaryk University Faculty ofScience Brno Czech Republic 2008

[20] L Van Valen ldquoA new evolutionary lawrdquo Evolutionary Theoryvol 1 pp 1ndash30 1973

[21] J Jaenike ldquoAn hypothesis to account for the maintenance ofsex within populationsrdquo EvolutionaryTheory vol 3 pp 191ndash1941978

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

BioMed Research International 9

We found higher mtDNA haplotype diversity in the gyno-genetic form when compared to the sexual form Our resultsindicate that changes in the frequencies of mitochondrialgenotypes across years do not explain the observed temporalvariability in parasite infection Different patterns of parasiteinfection in sexual and gynogenetic forms of C gibelio werereported when comparisons were made within individualyears a finding which may potentially be associated withthe dynamics of host-parasite interactions However theobserved dynamics of parasite infection does not support theRQ hypothesis as we failed to find a significantly higher levelof parasite infection in the asexual form in any single year ofinvestigation With respect to future studies focused on thedynamics of host-parasite interactions we suggest the need toanalyse other genetic markers especially those related to hostimmunity

Conflicts of Interest

The authors declare no conflicts of interest regarding thepublication of this paper

Authorsrsquo Contributions

Andrea Simkova designed the study Tomas Pakosta analysedthe data Lukas Vetesnık collected fish material All authorswere involved in drafting and revising the manuscript Allauthors read and approved the final version of the manu-script

Acknowledgments

The study was funded by the Czech Science FoundationProject no P505120375 Andrea Simkova and TomasPakosta were partially funded by the Czech Science Founda-tion ECIP project no P50512G112 The authors are grate-ful to M Benovics L Gettova M L Cervenka Kicinja KKoukalova V Krasnovyd C Rahmouni and G Vagnerovafor their help with fish dissection and parasite collection andK Civanova and S Stierandova for their help with molecularanalyses They would like to thank K Halacka from theInstitute of Vertebrate Biology Academy of Sciences of theCzech Republic for his help with fish sampling They alsothank Matthew Nicholls for English language revision of themanuscript

References

[1] J M Smith The Evolution of Sex Cambridge University PressCambridge UK 1978

[2] G BellTheMasterpiece of Nature the Evolution and Genetics ofSexuality University of California Press Berkeley Calif USA1982

[3] D K Lamatsch andM Stock ldquoSperm-dependent parthenogen-esis and hybridogenesis in teleost fishesrdquo in Lost Sex I SchonK Martens and P Dijk Eds pp 399ndash432 Springer DordrechtNetherlands 2009

[4] F W Allendorf ldquoEcological and Genetic Effects of Fish Intro-ductions Synthesis and Recommendationsrdquo Canadian Journalof Fisheries andAquatic Sciences vol 48 no S1 pp 178ndash181 1991

[5] P B Moyle and T Light ldquoFish invasions in California Doabiotic factors determine successrdquo Ecology vol 77 no 6 pp1666ndash1670 1996

[6] I G Cowx ldquoIntroduction of fish species into european freshwaters Economic successes or ecological disasters rdquo BFPP -Bulletin Francais de la Peche et de la Protection des MilieuxAquatiques no 344-345 pp 57ndash77 1997

[7] S Lusk V Luskova and K Halacka ldquoPrussian carp ndash 25 yearssince its natural introductionrdquo in Proceedings of the 3rd CzechConference of Ichthyology pp 135ndash140 1998

[8] J A Szczerbowski P M Banarescu and H J Poepke ldquoTheFreshwater Fishes of Europe Cyprinidae 2 Part III Carassiusto Cyprinusrdquo AULA-Verlag vol 5 no 3 pp 5ndash41 2002

[9] S Lusk and V Luskova ldquoInvasive fish species in the CzechRepublicrdquo in Proceedings of the 8th Czech Conference of Ichthy-ology pp 135ndash140 Czech 2005

[10] E Garcıa-Berthou C Alcaraz Q Pou-Rovira L Zamora GCoenders and C Feo ldquoIntroduction pathways and establish-ment rates of invasive aquatic species in Europerdquo CanadianJournal of Fisheries and Aquatic Sciences vol 62 no 2 pp 453ndash463 2005

[11] S Lusk V Barus andV Vesely ldquoOn the occurrence ofCarassiusauratus in the Morava river drainage areardquo Folia Zoologica vol4 no 26 pp 377ndash381 1977

[12] S Lusk V Luskova and L Hanel ldquoAlien fish species in theCzech Republic and their impact on the native fish faunardquo FoliaZoologica vol 59 no 1 pp 57ndash72 2010

[13] S Lusk J Kosco V Luskova K Halacka and P Kosuth ldquoAlienfish species in the floodplains of theDyje and the Bodrog riversrdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 199ndash205 2004

[14] S Lusk V Barus and A Kirka ldquoCurrent spreading andimportance of the giebel (Carassius auratus gibelio Bloch) inCzechoslovakiardquo Czech Journal of Animal Science vol 25 no11 pp 871ndash878 1980 (Czech)

[15] V Barus and O Oliva Fauna of Czech Republic and SlovakRepublic Petromyzontes and Osteichtyes Academia PragueCzech Republic 1995

[16] V Luskova K Halacka L Vetesnık and S Lusk ldquoChanges ofploidy and sexuality status of lsquoCarassius auratusrsquo populationsin the drainage area of the River Dyje (Czech Republic)rdquoEcohydrology amp Hydrobiology vol 4 no 2 pp 165ndash171 2004

[17] V Luskova S Lusk K Halacka and L Vetesnık ldquoCarassiusauratus gibeliomdashThe most successful invasive fish in waters ofthe Czech RepublicrdquoRussian Journal of Biological Invasions vol1 no 3 pp 176ndash180 2010

[18] L Vetesnık K Halacka V Luskova and S Lusk ldquoErythrocyteprofile of diploid and triploid silver crucian carp (Carassiusauratus)rdquo Acta Veterinaria Brno vol 75 no 2 pp 203ndash2072006

[19] I Papousek Molecular-genetic analyses of Carassius species incentral Europe [PhD thesis] Masaryk University Faculty ofScience Brno Czech Republic 2008

[20] L Van Valen ldquoA new evolutionary lawrdquo Evolutionary Theoryvol 1 pp 1ndash30 1973

[21] J Jaenike ldquoAn hypothesis to account for the maintenance ofsex within populationsrdquo EvolutionaryTheory vol 3 pp 191ndash1941978

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

10 BioMed Research International

[22] C M Lively C Craddock and R C Vrijenhoek ldquoRed Queenhypothesis supported by parasitism in sexual and clonal fishrdquoNature vol 344 no 6269 pp 864ndash866 1990

[23] K P Lampert P Fischer and M Schartl ldquoMajor histocompat-ibility complex variability in the clonal Amazon molly Poeciliaformosa Is copy number less important than genotyperdquoMolec-ular Ecology vol 18 no 6 pp 1124ndash1136 2009

[24] C M Lively and M F Dybdahl ldquoParasite adaptation to locallycommon host genotypesrdquo Nature vol 405 no 6787 pp 679ndash681 2000

[25] H Hakoyama T Nishimura N Matsubara and K IguchildquoDifference in parasite load and nonspecific immune reactionbetween sexual and gynogenetic forms of Carassius auratusrdquoBiological Journal of the Linnean Society vol 72 no 3 pp 401ndash407 2001

[26] H Hakoyama and Y Iwasa ldquoCoexistence of a sexual and anunisexual form stabilized by parasitesrdquo Journal of TheoreticalBiology vol 226 no 2 pp 185ndash194 2004

[27] J Kotusz M Popiolek P Drozd K De Gelas V Slechtovaand K Janko ldquoRole of parasite load and differential habitatpreferences in maintaining the coexistence of sexual and asex-ual competitors in fish of the Cobitis taenia hybrid complexrdquoBiological Journal of the Linnean Society vol 113 no 1 pp 220ndash235 2014

[28] A Simkova M Kosar L Vetesnık and M Vyskocilova ldquoMHCgenes and parasitism in Carassius gibelio a diploid-triploid fishspecies with dual reproduction strategiesrdquo BMC EvolutionaryBiology vol 13 article 122 2013

[29] A Simkova PHyrsl KHalacka andLVetesnık ldquoPhysiologicaland condition-related traits in the gynogenetic-sexualCarassiusauratus complex different investments promoting the coexis-tence of two reproductive formsrdquo BMC Evolutionary Biologyvol 15 article 438 2015

[30] L Vetesnık K Halacka and A Simkova ldquoThe effect of ploidyand temporal changes in the biochemical profile of gibel carp(Carassius gibelio) A cyprinid fish species with dual reproduc-tive strategiesrdquo Fish Physiology and Biochemistry vol 39 no 2pp 171ndash180 2013

[31] R Ergens and J Lom Causative Agents of Fish DiseasesAcademia Prague Czech Republic 1970

[32] G Malmberg ldquoOn the occurrence of Gyrodactylus on Swedishfishesrdquo Skrifter utgivna av Sodra Sveriges Fiskeriforening pp 19ndash76 1957 (Swedish)

[33] M Flajshans ldquoA model approach to distinguish diploid andtriploid fish by means of computer-assisted image analysisrdquoActa Veterinaria Brno vol 66 no 2 pp 101ndash110 1997

[34] K Halacka and V Luskova ldquoPolyploidy in Carassius auratus inthe lower reaches of the riverDyje - determination using the sizeof erythrocyte nucleirdquo in Proceedings of the 4th Czech Ichthy-ological Conference Book of Abstracts pp 106ndash109 ResearchInstitute of Fish Culture and Hydrobiology Vodnany CzechRepublic 2000

[35] J Sambrook E F Fritsch and TManiatisMolecular Cloning ALaboratoryManual University of Texas SouthWesternMedicalCenter Dallas Tex USA 1989

[36] L Vetesnık I Papousek K Halacka V Luskova and J MendelldquoMorphometric and genetic analysis of Carassius auratus com-plex from an artificial wetland in Morava River floodplainCzech Republicrdquo Fisheries Science vol 73 no 4 pp 817ndash8222007

[37] I Papousek L Vetesnık K Halacka V Luskova M Humpland J Mendel ldquoIdentification of natural hybrids of gibel carp

Carassius auratus gibelio (Bloch) and crucian carp Carassiuscarassius (L) from lower Dyje River floodplain (Czech Repub-lic)rdquo Journal of Fish Biology vol 72 no 5 pp 1230ndash1235 2008

[38] H Liu C-S Tzeng and H-Y Teng ldquoSequence variations inthe mitochondrial DNA control region and their implicationsfor the phylogeny of the Cypriniformesrdquo Canadian Journal ofZoology vol 80 no 3 pp 569ndash581 2002

[39] K Tamura G Stecher D Peterson A Filipski and S KumarldquoMEGA6 Molecular Evolutionary Genetics Analysis version60rdquoMolecular Biology and Evolution vol 30 no 12 pp 2725ndash2729 2013

[40] M Clement D Posada and K A Crandall ldquoTCS A computerprogram to estimate gene genealogiesrdquo Molecular Ecology vol9 no 10 pp 1657ndash1659 2000

[41] W D Hamilton ldquoSex versus Non-Sex versus Parasiterdquo Oikosvol 35 no 2 p 282 1980

[42] W D Hamilton R Axelrod and R Tanese ldquoSexual reproduc-tion as an adaptation to resist parasites (A review)rdquo Proceedingsof the National Acadamy of Sciences of the United States ofAmerica vol 87 no 9 pp 3566ndash3573 1990

[43] M Tobler and I Schlupp ldquoParasites in sexual and asexualmollies (Poecilia Poeciliidae Teleostei) A case for the RedQueenrdquo Biology Letters vol 1 no 2 pp 166ndash168 2005

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom