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Ticks, Fleas, and Harboured Pathogens from Dogs and Cats in Cyprus

Authors:
Anastasia Diakou at Aristotle University of Thessaloniki
Anastasia Diakou
Dimitra Sofroniou
Dimitra Sofroniou
Dimitra Sofroniou
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Barbara Paoletti at Università degli Studi di Teramo
Barbara Paoletti
Androniki Tamvakis at University of the Aegean
Androniki Tamvakis
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Abstract

Ticks and fleas are blood-sucking ectoparasites that cause irritation and anaemia to their hosts and act as vectors of pathogens (vector-borne pathogens, VBPs) of relevance for animal and human health. In the present study, tick and flea species in dogs and cats from Cyprus were recorded and VBPs were detected in the collected specimens. Ectoparasites were collected from 220 animals (161 dogs and 59 cats), and a questionnaire including demographic, clinical, and other information was filled out for each animal. The ectoparasites were morphologically identified and the detection of VBPs was performed by PCR-coupled sequencing. Rhipicephalus sanguineus sensu lato was found on 108 dogs and 13 cats, and Ixodes gibbosus on 2 dogs. Ctenocephalides felis was the predominant flea species (on 62 dogs and 45 cats), while one dog and one cat were infested by Ctenocephalides canis and Echid-nophaga gallinacea, respectively. The VBPs in ticks were Anaplasma platys, Rickettsia massiliae, Rickettsia conorii, Rickettsia felis, Hepatozoon felis and Hepatozoon canis, while Rickettsia felis, Rickettsia sp., Bartonella koehlerae, Bartonella clarridgeiae, and Bartonella henselae were recorded in fleas. Statistical analysis (chi-square test and multiple univariate generalized linear model) showed that animals up to 6 months of age were less likely to be infested with ticks than older animals, but more likely to be infested with fleas. Ticks were more prevalent in sheltered than in owned animals, while the odds ratio of flea presence was higher in owned animals than those living in shelters. The present study is the first investigation on the occurrence of ticks and fleas in dogs and cats from Cyprus, showing the presence of different VBPs in these important ectoparasites. The results point out the importance of systematic ectoparasite control in dogs and cats.
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Pathogens2022,11,1403.https://doi.org/10.3390/pathogens11121403www.mdpi.com/journal/pathogens
Article
Ticks,Fleas,andHarbouredPathogensfromDogs
andCatsinCyprus
AnastasiaDiakou
1,
*,DimitraSofroniou
2
,BarbaraPaoletti
3
,AndronikiTamvakis
4
,StanislavKolencik
5
,
DimitrisDimzas
1
,SimoneMorelli
3
,MarikaGrillini
6
andDonatoTraversa
3
1
LaboratoryofParasitologyandParasiticDiseases,SchoolofVeterinaryMedicine,FacultyofHealth
Sciences,AristotleUniversityofThessaloniki,54124Thessaloniki,Greece
2
IndependentResearcher,Love4PetsOmoniasAve.26,25088Lemesos,Cyprus
3
FacultyofVeterinaryMedicine,TeachingVeterinaryHospital,UniversityofTeramo,64100Teramo,Italy
4
LaboratoryofEcologyandSystemDynamics,DepartmentofMarineSciences,UniversityoftheAegean,
81100Mytilene,Greece
5
DepartmentofBiology,UniversityofNevadaReno,Reno,NV89557,USA
6
DepartmentofAnimalMedicine,ProductionandHealth,UniversityofPadua,35020Legnaro,Italy
*Correspondence:diakou@vet.auth.gr
Abstract:Ticksandfleasareblood‐suckingectoparasitesthatcauseirritationandanaemiatotheir
hostsandactasvectorsofpathogens(vector‐bornepathogens,VBPs)ofrelevanceforanimalandhu‐
manhealth.Inthepresentstudy,tickandfleaspeciesindogsandcatsfromCypruswererecorded
andVBPsweredetectedinthecollectedspecimens.Ectoparasiteswerecollectedfrom220animals(161
dogsand59cats),andaquestionnaireincludingdemographic,clinical,andotherinformationwas
filledoutforeachanimal.TheectoparasitesweremorphologicallyidentifiedandthedetectionofVBPs
wasperformedbyPCR‐coupledsequencing.Rhipicephalussanguineussensulatowasfoundon108
dogsand13cats,andIxodesgibbosuson2dogs.Ctenocephalidesfeliswasthepredominantfleaspecies
(on62dogsand45cats),whileonedogandonecatwereinfestedbyCtenocephalidescanisandEchid‐
nophagagallinacea,respectively.TheVBPsintickswereAnaplasmaplatys,Rickettsiamassiliae,Rickettsia
conorii,Rickettsiafelis,HepatozoonfelisandHepatozooncanis,whileRickettsiafelis,Rickettsiasp.,Bartonella
koehlerae,Bartonellaclarridgeiae,andBartonellahenselaewererecordedinfleas.Statisticalanalysis(chi‐
squaretestandmultipleunivariategeneralizedlinearmodel)showedthatanimalsupto6monthsof
agewerelesslikelytobeinfestedwithticksthanolderanimals,butmorelikelytobeinfestedwith
fleas.Ticksweremoreprevalentinshelteredthaninownedanimals,whiletheoddsratiooffleapres‐
encewashigherinownedanimalsthanthoselivinginshelters.Thepresentstudyisthefirstinvesti‐
gationontheoccurrenceofticksandfleasindogsandcatsfromCyprus,showingthepresenceof
differentVBPsintheseimportantectoparasites.Theresultspointouttheimportanceofsystematic
ectoparasitecontrolindogsandcats.
Keywords:ectoparasites;epidemiology;petanimals;vector‐bornepathogens
1.Introduction
Ticksandfleasareblood‐suckingarthropods,infestingseveralvertebrates,among
themdogsandcats.Theyhavebeenextensivelystudiedbecauseoftheirdirectclinical
impactonanimals,thepathogenstheytransmit,andtheirrelevanceinhumanhealth[1,2].
Theseectoparasitescancausediscomfortandmayseverelyimpactthehealthandwell‐
beingofdogsandcats.Tickscausenuisance,anaemia,irritation,cutaneouslesionswith
inflammationandeosinophilicaggregation,secondaryinfectionsoccasionallyleadingto
abscessesorevenpyaemia,andtoxicosis(tickparalysis).Fleascausesevereirritation,
pruritusandself‐woundformation,bloodlossandanaemia,andflea‐associatedallergic
dermatitis[3–5].Ticksandfleasmayalsotransmitvariousvector‐bornepathogens(VBPs)
Citation:Diakou,A.;Sofroniou,D.;
Paoletti,B.;Tamvakis,A.;
Kolencik,S.;Dimzas,D.;Morelli,S.;
Grillini,M.;Traversa,D.Ticks,Fleas,
andHarbouredPathogensfrom
DogsandCatsinCyprus.Pathogens
2022,11,1403.https://doi.org/
10.3390/pathogens11121403
AcademicEditor:
LawrenceS.Young
Received:5November2022
Accepted:21November2022
Published:23November2022
Publisher’sNote:MDPIstaysneu‐
tralwithregardtojurisdictional
claimsinpublishedmapsandinstitu‐
tionalaffiliations.
Copyright:©2022bytheauthors.Li‐
censeeMDPI,Basel,Switzerland.
Thisarticleisanopenaccessarticle
distributedunderthetermsandcon‐
ditionsoftheCreativeCommonsAt‐
tribution(CCBY)license(https://cre‐
ativecommons.org/licenses/by/4.0/).
Pathogens2022,11,14032of13
totheirhosts,manyofwhicharezoonotic.Pathogenstransmittedbytickstodogsand
catsincludemostlyprotozoa(e.g.Babesiaspp.,Hepatozoonspp.,Cytauxzoonspp.)andbac‐
teria(Rickettsiaspp.,Ehrlichiaspp.,Anaplasmaspp.,Coxiellaspp.,Borreliaspp.).Fleasare
vectorsofBartonellaspp.,Rickettsiafelis,andYersiniapestis,andarealsoanintermediate
hostofthecestodesDipylidiumcaninumandHymenolepisdiminuta,andthenematode
Acanthocheilonemareconditum[3,6–8].
Specificdrivers,e.g.,climatechangeandglobalwarming,destructionofwildhabi‐
tatsforagricultureintensification,landscapemodification,poorecosystemprotection,
andincreaseinpettravelhaveasignificantimpactontheepidemiologyandtheincreas‐
ingoccurrenceofectoparasites[6].Consequently,theaffiliatedVBPsandassociateddis‐
easesareexpectedtoexpand,emerge,orre‐emergeinmanyareas[9].Knowledgeofthe
currentepidemiologyofticks,fleas,andtransmittedpathogensisstillscantinmanyareas
ofEuropeandtheirdistributionandoccurrenceareconstantlychangingovertime[10].
InCyprus,someinvestigationsonticksandtick‐bornepathogenshavebeencon‐
ductedinthepast[11–16],whiledataonfleasandflea‐bornepathogensarelimitedto
onlyrats,foxes,andhares[14,17,18].Therefore,theaimofthepresentstudywas(i)to
investigatetheinfestationbyticksandfleasindogsandcatsfromCyprus;(ii)todetect
thepresenceofVBPsintheseectoparasites;and(iii)toassociatefindingswithdifferent
possibleriskfactors,inordertoupdateandenrichknowledgeabouttheepidemiologyof
theseimportantectoparasites.
2.MaterialsandMethods
2.1.AnimalsandEctoparasiteCollection
Thesurveywasconductedon220animals(161dogsand59cats),livinginfivedis‐
trictsofCyprus,i.e.,Ammochostos,Larnaca,Lemesos,Lefkosia,andPaphos(Figure1),
andpresentedtoaprivateveterinaryclinicinLimassolforroutineclinicalexaminations
(e.g.,vaccination,castration,investigationofclinicalcondition,injury).Ectoparasiteswere
detectedbyfurandskininspectionandbycombingwithastainless‐steelfleacomb.The
ectoparasiteswerecollectedbyentomologicalforceps,storedinEppendorftubescontain‐
ing70ethanol,andtaggedwithanindividualcode.Foreachanimalincludedinthesur‐
vey,aquestionnairewasfilledout,withinformationaboutage,sex,countrydistrict,life‐
style,lastectoparasiticideadministration,thereasonforthevisit,andclinicalandlabora‐
toryfindings.
Figure1.ThemapofCyprusandthedistrictsfromwhichthesampledanimalsoriginated.
Pathogens2022,11,14033of13
2.2.IdentificationofEctoparasites
ThecollectedectoparasitesweretransferredtotheLaboratoryofParasitologyandPar‐
asiticDiseases,SchoolofVeterinaryMedicineoftheAristotleUniversityofThessaloniki.
Theectoparasiteswereexaminedunderastereomicroscope(8×–64×)andalightmicroscope
(100×,400×)foridentificationbasedontheirmorphologicalcharacteristics[19–21].
2.3.DetectionofVBPs
Afteridentification,ectoparasitespecimensweretransferredtotheLaboratoryof
ParasitologyoftheFacultyofVeterinaryMedicine,UniversityofTeramo,forthedetec‐
tionofVPBsbymolecularmethods.
Ticksandfleaswereexaminedinpooledsamplesperanimal,intogroupsofoneto
fiveindividuals.Overall,122pooledticksamplesand111pooledfleasampleswereex‐
amined,excludinghighlyengorgedtickspecimenstoavoidexcessnucleicacidsofverte‐
bratehostorigin.TheectoparasitepoolswerehomogenizedbeforeDNAextraction.
Briefly,thespecimensweretakenfromthe70ethanolsolution,air‐dried,andmechani‐
callycrushedina1.5mlsafe‐locktubewithsterilepestles.Thehomogenateswereincu‐
batedwithproteinaseKsolutionovernightat56°Candtotalnucleicacidswereextracted
fromthesehomogenatesinaccordancewiththemanufacturer’sinstructions(ExgeneTis‐
sueSV,GeneAll,SouthKorea).Inticks,Anaplasmaspp./Ehrlichiaspp.,Babesiaspp.,Bar‐
tonellaspp.,Rickettsiaspp.,andHepatozoonspp.,andinfleas,Bartonellaspp.andRickettsia
spp.,weredetectedbypolymerasechainreaction(PCR).Afragmentofthe18SrRNAgene
ofAnaplasma/Hepatozoonspp.andBabesiaspp.,apartialsequenceofthe16S–23SrRNA
intergenicspeciesregion(ITS)ofBartonellaspp.,andafragmentoftherickettsialouter
membraneproteinA(ompA)genewereamplifiedusingprimersandprotocolsdescribed
previously[22–25].TheprimersusedfortheamplificationofthetargetedDNAareshown
inTable1.AllamplificationsincludedapositivecontrolcontaininggenomictargetDNA
andanegativecontrolwithoutDNA.PCRproductswerevisualizedunderUVillumina‐
tionafterelectrophoresismigrationona1.8%agarosegel.PCRproductsweresequenced
inonedirection,usingthesameprimersasthoseusedforDNAamplification.Sequences
werecomparedforsimilaritytosequencesinGenBank,usingtheBLASTprogramhosted
byNCBI,NationalInstitutesofHealth,USA(http://www.ncbi.nlm.nih.gov,accessedon1
August2022).
Table1.PrimersusedforthedetectionofVBPsinectoparasitesofdogsandcatsfromCyprusand
correspondingreferences(Ref).
PrimerPathogenTargetgeneNucleotideSequences(5’‐3’)Product
Size(bp)Ref
Rrl9O.70pRickettsia190kDaanti‐
gen
ATGGCGAATATTTCTCCAAAA~532[22]
Rrl9O.602nAGTGCAGCATTCGCTCCCCCT
325sBartonella16S‐23SrRNA
ITS
CTTCAGATGATGATCCCAAGCCTTYTGGCG~600[23]
1100asGAACCGACGACCCCCTGCTTGCAAAGCA
PiroABabesia18SrRNAAATACCCAATCCTGACACAGGG400[24]
PiroBTTAAATACGAATGCCCCCAAC
EHR16SDAnaplasma/Ehr‐
lichia18SrRNAGGTACCYACAGAAGAAGTCC345[24]
EHR16SRTAGCACTCATCGTTTACAGC
TabarFHepatozoon18SrRNACCAGCAGCCGCGGTAATTC373[25]
TabarRCTTTCGCAGTAGTTYGTCTTTAACAAATCT
2.4.StatisticalAnalysis
Theoccurrenceoffleasandticksondogsandcatswasevaluatedinrelationtofactors
expressingdemographicdetails(gender,age),status(ownedorsheltered),andprevious
Pathogens2022,11,14034of13
treatments(timepassedsincethelastdosing).Moreover,theexistenceofVBPsintheec‐
toparasiteswasassociatedwithadditionalfactors:geographicregion,thestatusofthe
animal(ownedorsheltered),andclinicalexaminationorlaboratoryfindingsassociated
withdisease(e.g.,anorexia,weightloss,eyelesions,neurologicalsigns,positivein‐clinic
diagnostictestforinfectiousdiseases).Thechi‐squaretestofindependencewasusedto
assesstheeffectoftheabovefactorsontheoccurrenceofectoparasitesandtheexistence
ofVBPs,respectively.Thesignificantfactorsdefinedbythechi‐squaretestwerethenen‐
teredintoamultipleunivariategeneralizedlinearmodel(GLM)fordeterminingtheir
combinedeffectontheoccurrenceofectoparasites[26].Theoddsratioswiththeircorre‐
spondingconfidenceintervals(C.I.)wereusedtocomparetheproportionoftheoccur‐
renceofeachectoparasiteamongthefactorgroups.Theinformationcollectedthroughthe
questionnaires,abouttheveterinaryproductusedonsomeoftheanimals,wasnotin‐
cludedinthestatisticalanalysisowingtomissingorunreliabledata.Thestatisticalanal‐
ysiswasimplementedusingtheRpackageversion[27]andtheRcmdrpackage[28].
3.Results
3.1.StudyAnimals
ThedemographicsandotherdetailsoftheexaminedanimalsareshowninTable2.
Table2.Recordeddataforthedogsandcats(n=220)withectoparasitesexaminedinCyprus.
FactorDogs(n=161)Cats(n=59)
StatusOwner/Shelter134/2751/8
Region
Lefkosia323
Lemesos9142
Larnaca286
Paphos84
Amochostos24
SexMale/Female81/8023/36
Age
<6months2317
6–≤12months1314
>1–≤7years9325
>7years323
Lasttreatmentfor
ectoparasites
≤1month172
1–≤3months176
>3–≤6months113
>6–≤12months220
>12months9448
Reasonforvisitor
findingsDisease/Other88/7334/25
3.2.Ectoparasites
Fromatotalof161dogswithectoparasites,98and51hadticksorfleasonly,respec‐
tively,while12hadmixedtickandfleainfestation.Accordingly,intotal,110(68.3%)dogs
wereinfestedwithticksand63(39.1%)hadfleas,includingbothsingleandmixedinfec‐
tions.Fromatotalofthe59catsinfestedwithectoparasites,9hadonlyticks;45hadonly
fleas;3hadticksandfleas;1hadfleasandlice;and1hadamixedinfestationwithticks,
fleas,andlice.Intotal,13(22%)catswereinfestedwithticks,50(84.7%)withfleas,and2
(3.4%)withlice,includingbothsingleandmixedinfections(Tables3and4).
Pathogens2022,11,14035of13
Table3.Number(n)ofanimals(dogsorcats)inCyprus,infestedwithdifferenttypesofectopara‐
sites,withthecorrespondingconfidenceinterval(C.I.)oftheoccurrencepercentage.
AnimalSpecies
(SampleSize)
Ticks
n(%C.I.)
Fleas
n(%C.I.)
TicksandFleas
n(%C.I.)
FleasandLice
n(%C.I.)
Ticks,Fleas,andLice
n(%C.I.)
Dogs(n=161)98(60.8±7.7)51(31.7±6.7)12(7.5±3.1)00
Cats(n=59)9(15.3±7.0)45(76.3±12.2)3(5.1±3.3)1(1.7±1.4)1(1.7±1.4)
Twodifferenttickspecieswereidentified:Rhipicephalussanguineussensulato(s.l.)on
108dogsand13cats,andIxodesgibbosuson2dogs.Themostabundantfleaspecieswas
Ctenocephalidesfelis,foundon62dogsand45cats,whileCtenocephalidescanisandEchid‐
nophagagallinaceawerefoundononedogandonecat,respectively.Themixedinfestations
included10dogsand4catswithR.sanguineuss.l.andC.felis;twodogswithI.gibbosus
andC.felis;onecatwithR.sanguineuss.l.,C.felis,andthelouseFelicolasubrostratus;and
onecatinfestedwithE.gallinaceaandF.subrostratus(Table4).
Table4.SpeciesidentificationofticksandfleasandmixedinfectionsindogsandcatsfromCyprus.
AnimalSpecies
(SampleSize)
Rhipicephalussan‐
guineuss.l.
Ixodesgib‐
bosusCtenocephalidesfelisCtenocephalidesca‐
nis
Echidnophagagal‐
linacea
Dogs(n=161)108122621,210
Cats(n=59)133,40453,4015
1TendogswithmixedinfestationbyR.sanguineusandC.felis;22dogswithmixedinfestationbyI.
gibbosusandC.felis;34catswithmixedinfestationbyR.sanguineusandC.felis;4acatwithamixed
infestationbyR.sanguineus,C.felis,andFelicolasubrostratus;5acatwithamixedinfestationbyE.
gallinaceaandthelouseF.subrostratus.
3.3.DetectionofVBPs
Intotal,233ectoparasitesamples(122tickand111fleasamples)wereexaminedfor
thedetectionofVBPs.Inthecaseofmultipleticksorfleaspecimensperanimal,apooled
sample(perectoparasitetypeandperanimal)wasprepared.VBPs’detectionbyPCRwas
notpossibleforonetickandtwofleasamplesowingtoaninsufficientornotsuitableDNA
sample.Overall,32(14.5%)animalswereinfestedwithectoparasitesthatharbouredone
ormoreVBPs,whereas35(15%)ectoparasitepoolsampleswerepositiveforVBPs,be‐
cause,inthreecases(twodogsandonecat)withamixedinfestationbyR.sanguineuss.l.
andC.felis,VBPswerefoundinbothticksandfleas.
TheDNAofsixdifferentpathogenswasdetectedinticks,i.e.,Anaplasmaplatys,Rick‐
ettsiamassiliae,Rickettsiaconorii,Rickettsiafelis,Hepatozoonfelis,andHepatozooncanis,while
noBabesiaspp.wasfoundintheexaminedspecimens.TheDNAoffivedifferentVBPs
wasdetectedinfleas,i.e.,Rickettsiafelis,Rickettsiasp.,Bartonellakoehlerae,Bartonellaclar‐
ridgeiae,andBartonellahenselae.Detailsaboutthespeciesandnumberofanimalsinthe
ectoparasitesofwhichtheseVBPsweredetectedareshowninTable5.
Table5.Vector‐bornepathogens(VBPs)detectedin122tickand111fleapooledsamples(perec‐
toparasitetypeandperanimal)collectedfromdogsandcatsinCyprus.
Animal
Species
VBPsinTicksVBPsinFleas
A
.pR.mR.
c
R.
f
H.cH.
f
R.
f
R.sp.B.kB.
c
B.h
Dogs(n)3101‐ 313412‐
Cats(n) 2‐ 1‐ 15‐ ‐ 1
Total312113284121
n=numberofanimalsintheectoparasitesofwhichthepathogenwasfound,A.p.=Anaplasma
platys;R.m=Rickettsiamassiliae;R.c=Rickettsiaconorii;R.f=Rickettsiafelis;H.f=Hepatozoonfelis;
H.c=Hepatozooncanis;R.sp.=Rickettsiasp.;B.k=Bartonellakoehlerae;B.c=Bartonellaclarridgeiae;B.
h=Bartonellahenselae.
Pathogens2022,11,14036of13
SequencingofPCRproductsandBLASTanalysisrevealedsimilaritiesoftheherein
detectedVBPswithDNAsequencespublishedinGenBank,asshowninTable6.
Table6.Vector‐bornepathogens(VBPs)detectedinticksandfleasfromdogsandcatsinCyprus,
andtheirsimilaritywithGenBankentries.
VBP(nofSequencesAnalyzed)GenBankAccessionNumberSimilarity
Anaplasmaplatys(n=3)JX392984.199%
Rickettsiamassiliae(n=12)MW026209.197–99%
Rickettsiafelis(n=9)KP318094.196–99%
Hepatozoonfelis(n=2)KY649442.1100%
Hepatozooncanis(n=3)MK645969.197–100%
Rickettsiaconorii(n=1)AE006914.197%
Rickettsiasp.(n=4)MF134884.196–99%
Bartonellakoehlerae(n=1)MT095046.198%
Bartonellaclarridgeiae(n=2)EU589237.196%
Bartonellahenselae(n=1)KT314216.1100%
3.4.StatisticalAnalysis
Chi‐squaretestofindependenceshowedthatneitherticknorfleapresencewasre‐
latedtothetimepassedsincethelastectoparasitictreatment(χ2=3.68,df=4,p>0.05for
ticksandχ2=3.54,df=4,p>0.05forfleas)ortheanimal’ssex(χ2=0.60,df=1,p>0.05for
ticksandχ2=0.02,df=1,p>0.05forfleas).Ontheotherhand,theoccurrenceofectopar‐
asiteswasassociatedwiththeageofthehost(χ2=27.19,df=3,p<0.001forticksandχ2=
20.90,df=3,p<0.001forfleas)andtheir“ownedorsheltered”status(χ2=14.99,df=1,p
<0.001forticksandχ2=16.34,df=1,p<0.001forfleas)(Table7).
Table7.Chi‐squaretestofindependenceshowingassociationsbetweentheoccurrenceofectopar‐
asitesandvariousfactorsrecordedforeachanimal.
TicksFleas
VariablePositiveNegativep‐valuePositiveNegativep‐value
Lasttreatment 0.451 0.471
≤1month14(73.7%)5(26.3%) 6(31.6%)13(68.4%)
>1–3months12(52.2%)11(47.8%) 13(56.5%)10(43.5%)
>3–6months6(42.9%)8(57.1%) 8(57.1%)6(42.9%)
>6–12months13(59.1%)9(40.9%) 11(50.0%)11(50.0%)
>12months78(54.9%)64(45.1%) 75(52.8%)67(47.2%)
Sex 0.438 0.875
Male61(58.7%)43(41.3%) 54(51.9%)50(48.1%)
Female62(53.4%)54(46.6%) 59(50.9%)57(49.1%)
Agecategory 0.000* 0.000*
<6months13(32.5%)27(67.5%) 30(75.0%)10(25.0%)
6–12months7(25.9%)20(74.1%) 20(74.1%)7(25.9%)
>1–7years81(68.6%)37(31.4%) 49(41.5%)69(58.5%)
>7years22(62.9%)13(37.1%) 14(40.0%)21(60.0%)
Status 0.000* 0.000*
Owned93(50.3%)92(49.7%) 106(57.3%)79(42.7%)
Sheltered30(85.7%)5(14.3%) 7(20.0%)28(80.0%)
*Statisticallysignificantfactor,p<0.001.
TheinvestigationoftheassociationbetweenVBPs’occurrenceandvariousfactors
showedthatVBPs’detectionwasnotassociatedwithclinicalsignsorfindingsofdisease
Pathogens2022,11,14037of13
(χ2=2.42,df=1,p>0.05),theanimals’“ownedorsheltered”status(χ2=1.06,df=1,p>
0.05),ortheregionofliving(χ2=3.62,df=4,p>0.05)(Table8).
Table8.Contingencytableswithchi‐squaretestresultsbetweenVBPs’existenceandotherfactors.
VBPsp‐Value
VariablePositiveNegative
Signs/findings 0.120
Disease21(17.8%)97(82.2%)
Other10(10.3%)87(89.7%)
Status 0.304
Owned24(13.3%)156(86.7%)
Sheltered7(20.0%)28(80.0%)
Region 0.460
Ammochostos2(40.0%)3(60.0%)
Larnaca3(9.4%)29(90.6%)
Lemesos18(13.7%)113(86.3%)
Lefkosia6(17.1%)29(82.9%)
Paphos2(16.7%)10(83.3%)
Theagecategoryandthe“ownedorsheltered”statuswerefurtheranalysedfortheir
combinedeffectontheoccurrenceofticksorfleasusingmultipleGLM(Table9).The
analysisshowedthatanimalsupto6monthsandthosebetween6and12monthshadthe
samelikelihoodtobeinfestedbyticksorfleas(multipleGLMp‐values>0.05).However,
younganimalshadahigherlikelihoodofbeinginfestedwithfleas,whereasolderanimals
hadahigherlikelihoodofbeinginfestedwithticks.Indeed,animalsupto6monthswere
0.26and0.27timeslesslikelytobeinfestedwithticksthananimals1to7yearsorolder,
respectively.Animalsupto6monthswere3.59and4.88timesmorelikelytobeinfested
withfleasthananimalsfrom1to7yearsandthoseolderthan7years,respectively(mul‐
tipleGLMp‐value<0.01).Thestatus(ownedorsheltered)oftheanimalwasalsofound
toberelatedtothepresenceofectoparasites(multipleGLMp‐value<0.01).Tickswere
fivetimes(i.e.theinverseof0.2oddsratioshowninTable9)morelikelytobefoundon
shelteredanimalsthanownedanimals,whiletheoddsratiooffleapresencewas4.84
timeshigherinownedanimalsthaninthoselivinginshelters.
Table9.Assessmentofriskfactorsofectoparasites’occurrenceincludingtheresultsofthemulti‐
plegeneralizedlinearmodel(GLM).
TicksFleas
VariableOddsRatio95%CIGLM
p‐valueOddsRatio95%CIGLM
p‐Value
Agecategory
<6mvs.6–12m 1.40(0.47,4.43)0.5571.05(0.32,3.30)0.928
vs.1–7years0.26(0.11,0.55)0.001*3.59(1.61,8.54)0.002*
vs.>7years0.27(0.10,0.67)0.007*4.88(1.83,13.82)0.001*
Status
Ownedvs.Sheltered0.20(0.06,0.52)0.002*4.84(2.04,12.91)0.001*
*MultipleGLMp‐value<0.01,identifyingariskfactor.
4.Discussion
Cyprus,anislandcountryintheEasternMediterraneanSea,isacosmopolitanhub
andacentreoftourism,market,education,andotheractivities,whichreceivesagreat
numberofvisitorsthroughouttheyear.Ontheotherhand,Cyprushasalargenumberof
Pathogens2022,11,14038of13
dogsandcats,livingasownedpets,free‐roaming,orstrays.Asignificantnumberofani‐
malsheltersinthecountryareactivelyfacilitatingadoptionofstrayanimals,which,in
manycases,travelabroad,totheirnewhome,indifferentareasoftheworld.Inthiscon‐
text,investigatingandmonitoringpathogensthatmaybetransmittedlocallyorinremote
countriesviaticksandfleasisofgreatepidemiologicalimportance.
Thesubtropical–MediterraneanclimateofCypruswithmildwintersandwarmto
hotsummersisfavourabletoticksandfleas,becausetheirdevelopment,especiallythe
rateoftransitionfromonedevelopmentstagetothenext,whichinmostcasestakesplace
intheenvironment,istemperature‐dependent[29,30].Thepresentresultsareinlinewith
thefactthattickparasitismismorecommonindogsthanincats,whiletheoppositeis
trueforfleainfestations,probablybecauseofthedifferentbehaviourofdogsandcatsand
thedifferentbiologyoftheseectoparasites[1,31].
Theectoparasitespeciesidentifiedhereinhaveaworldwidedistributionandareprev‐
alentinSouthernEurope[32].Thepredominanttickspecies,R.sanguineuss.l.[33],also
madeupthemajority(89–92%)ofthetickscollectedfromdogsinearliersurveysinCyprus,
showinglimitedaffiliationtootherhostspecies(mouflons,foxes,hares,goats,sheep,and
bovines)[13,15].Itisathree‐hosttick,afactthatfacilitatesthetransmissionofVBPsfrom
animaltoanimalandisthevectorofmanyVBPs[4,34].Accordingly,6differentVBPswere
detectedin22outof120R.sanguineuss.l.samplesexaminedinthepresentstudy.
TheprevalenceofA.platys,theagentofcaninecyclicthrombocytopenia(CCT),varies
between0.4%and87.5%indifferentareasoftheworld[35].InCyprus,thisbacteriumhas
beendetectedonlyonceinadog[16].Anaplasmaplatysisarecognizedzoonoticagent[35],
andenrichinginformationonitsoccurrenceinareaswheredataarelackingisimportant.
ThepresentresultsconfirmthatthisVBPiscirculatingamongticksanddogsinCyprus.
EventhoughseropositivedogstoR.conoriiarehighlyprevalentinsouthernEurope
[9,36–39],usually,onlyasmallnumberoftheexaminedticksscorepositivelyinPCR[40–
42],whichisconsistentwiththepresentresults.Theinfectionindogsisusuallysubclini‐
cal,butinhumans,R.conoriiistheagentofMediterraneanspottedfever[43],thuscreation
ofepidemiologicalinformationisessential.Interestingly,R.massiliaewasthemostpreva‐
lentVBPinthepresentstudy.ItisconsideredanemergingpathogeninAfrica,Europe,
andtheUSA,incriminatedforseveralhumancaseswithclinicalsignssimilartoMediter‐
raneanspottedfever[44].Onthebasisofthepresentfindings,R.massiliaeisapossible
emergingpublichealththreatinCyprusandtheawarenesstowardsthisbacteriumshould
beincreased.
BothH.canisandH.feliswerefoundinticks,albeitatalowprevalence.InCyprus,H.
canishasbeenpreviouslyreportedindogs[45],whileH.felishasbeendetectedwithahigh
prevalence(37.9%)incats[46].Similarly,H.felisoccurswithahighprevalenceincatsin
otherEuropeanenzooticareas,reaching25.5%inGreece[47].Hepatozoonspeciescirculating
inEurope,i.e.,H.canisindogsandH.canis,H.felis,andHepatozoonsilvestrisincats,have
diversepathogenicpotentials.Althoughinfectionsareoftensubclinical,animalsmayde‐
velopseverediseasedependingonthespeciesorhaplotypeinvolved[47–50].
Tothebestoftheauthors’knowledge,thisisthefirstrecordofI.gibbosusondogsin
Cyprus.ItisoneofthemostcommonIxodesspeciesontheisland[15,31]anditwaspre‐
viouslyreportedonmouflons[51].Ixodesgibbosusisadaptedtowarmanddryclimates,
replacingIxodesricinusintheeasternMediterranean,whichislessresistanttosuchcon‐
ditions[31].FurtherinvestigationsintotheprevalenceandvectorialcapacityofI.gibbosus,
asthedominantIxodesspeciesinthearea,wouldbeofmerit.
Ctenocephalidesfelisisthevectorofimportantpathogens,includingB.henselaede‐
tectedhereinandpreviouslyreportedinratsandcatsofCyprus[6,46,52].Ontheother
hand,tothebestoftheauthors’knowledge,thepresentreportofB.koehleraeandB.clar‐
ridgeiaeisthefirstinthecountry.Bartonellaspp.areagentsofdiseaseinbothanimalsand
humans;forexample,B.henselaeistheagentofcat‐scratchdisease[53],thusconstantsur‐
veillanceofthepresenceoftheseVBPsindogs,cats,andectoparasitesispivotal.
Pathogens2022,11,14039of13
Rickettsiafelisistheagentofhumanflea‐bornespottedfeverandanemergingVBP
[6].InCyprus,ithasbeendetectedpreviouslyinC.felisfromrats[17].Thecatfleaisthe
primaryvectorofR.felis,butitisprobablyalsotransmittedbyotherfleaspecies,ticks,
andotherblood‐suckingarthropods[6,54]anditwasalsodetectedinR.sanguineuss.l.in
thepresentstudy.
Ctenocephalidescanis,thedogflea,islesscommonindogsthanC.felis[6].Accord‐
ingly,thisfleaspecieswasfoundonlyononedoginthepresentstudy.Nevertheless,in
someareas,C.canisisreportedtobemoreprevalentthanC.felis[55].Thedogfleamay
alsotransmitpathogensincludingR.felisandB.henselae;however,becauseofitslimited
abundancecomparedwiththecatflea,itsvectorialroleisconsideredinferior[56].
ThefleaE.gallinacea,alsoknownasthe“sticktightflea”,wasfoundononecat.This
speciesiscommononfowl,butitalsoinfestsmammals,mostfrequentlycats,probably
owingtobirdhunting[57].Itisafleaspeciesofbothveterinaryandmedicalimportance,
transmittingfowlviruses,Y.pestis,R.typhi,andD.caninum[58],whichrendersitanim‐
portanttargetforstudyandcontrol,despiteitslowfrequency.
AnincidentalfindingintheexaminationforticksandfleaswasthecatlouseF.subro‐
stratusontwocats.Catlousehasaworldwidedistributionandinfestationisoftenanin‐
dicationofapoorgeneralhealthconditionandlackofcare[59].Evenifoutofthescope
ofthepresentarticle,thisfindingisimportantasthecatlousehasbeenidentifiedasa
potentialintermediatehostofaDipylidiumspecies,geneticallydistantfromD.caninum,
infectinghyenas,dogs,andcats[60].
TheuseofompAgeneappearstobespecificanddiscriminatingforthespottedfever
groupRickettsiae,butsomeauthorsrecommendthatmultiplegenetargetsshouldbeused
togainanaccurateidentification[61].Thiscouldbethereasonthat,forafewisolates,iden‐
tificationonlytothegenuslevelwasfeasible(Table5).TheremainderofVBPsidentifiedin
thepresentstudyshowedavaryinglevelofsimilaritywithGeneBankdeposits,isolated
fromdifferenthostsandareasoftheworld(Table6).Itisworthnotingthatthedetectionof
Rickettsiaspp.DNA,mainlyinR.sanguineus,providesevidencethatthistickmaybeamong
themainvectorsofRickettsiaspp.inCyprus,accordingtopreviousstudies[40,62].
Thefindingthatyounganimals(<6months)weresignificantlylesslikelytobein‐
festedwithticks,butmorelikelytobeinfestedwithfleas,maybeattributedtothefact
thatyounganimalswillspendmostoftheirtimeinarestrictedenvironmentneartheir
home,incloseproximitytotheirmotherandsiblings,aconditionthatfavourshost‐to‐
hostfleatransmission[1].Thiscontrastswitholderanimalsthatspendmoretimeroaming
awiderareaoutdoors.Assuch,olderanimalsaremorelikelytocomeintocontactwith
ticks,explainingthefindingthatolderanimalsweresignificantlymorelikelytoharbour
ticksthanyounganimals.
Theactivitywithinawiderorrestrictedenvironmentmayalsobethereasonticks
weremoreprevalentonshelteredanimals,especiallyconsideringthatsomeofthemwere
introducedrecentlyandwerepreviouslyroaminginawiderareaoftheirregion.Accord‐
ingly,theoccurrenceoffleaswasmorefrequentinownedanimals,livinginaconfined/re‐
strictedenvironment(indoorsformostcats,indoorsor/andinthegardenfordogs),which
canoftenmaintainfleainfestation,comparedwiththoselivinginshelters.
Interestingly,theinfestationwasnotstatisticallyassociatedwiththetimethathad
passedsincethelastectoparasiticapplication.Thus,animalswitharecentectoparasite
treatmentwereatthesameriskofinfestationastherestoftheanimals.Althoughdrug
resistancedevelopmentinectoparasitesisaknownproblem[29,63],thelackofspecific
investigationintotheproductsusedandtheapplicationpracticesdoesnotallowfurther
evaluationofthisfinding.
5.Conclusions
Ticksandfleasareamajorconcernforpetowners,veterinarians,andmedicaldoctors
becauseoftheirclinicalimpactondogsandcatsandtheVBPstheytransmit.Theresults
ofthepresentstudyprovidenewknowledgeabouttheoccurrenceofticksandfleasin
Pathogens2022,11,140310of13
dogsandcatsfromCyprus,andthepathogensthattheseectoparasitesmaytransmit,cov‐
eringarelevantgapinknowledge.Companionanimalstravellingforadoption(com‐
monlyshelteredanimals)orwiththeirownersforvacationsmayfacilitatethespreading
ofVPBs[45].Thisriskislurking,particularlyinanimalmovementsfromandtoMediter‐
raneanareas,includingCyprus,asthispartofEuropeisconsideredamajorepidemiolog‐
icalhubforVBPs[47].Systematicectoparasitecontrolispivotalandaplethoraofveteri‐
naryproductsareavailableforthispurpose.Furthermore,theresearchintonewanimal
andenvironment‐friendlytoolsforcontrolisongoing,andeffectivebiologicalorbotani‐
cal‐basedcompoundsandvaccinesmayalsobeavailableinthefuture[64,65].
AuthorContributions:Conceptualization,A.D.andD.S.;methodology,A.D.,D.S.,B.P.,A.T.,S.K.,
D.D.,S.M.,andM.G.;datacuration,A.D.,D.S.,D.D.,S.K,andA.T.;writing—originaldraftprepa‐
ration,A.D.,B.P.,andA.T.;writing—reviewandediting,A.D.,S.M.,S.K,andD.T.;supervision,
A.D.andD.T.Allauthorshavereadandagreedtothepublishedversionofthemanuscript.
Funding:Thisresearchreceivednoexternalfunding.
InstitutionalReviewBoardStatement:Ethicalreviewandapprovalwerewaivedforthisstudyas
alloftheanimalsinvolvedwereclinicallyexaminedintheframeoftheirroutineveterinarycheck‐
upandnomedicalinvasiveprocedureswereperformed.
InformedConsentStatement:Informedconsentwasobtainedfromalloftheanimals’ownersfor
theuseofectoparasitesinresearch.
DataAvailabilityStatement:Notapplicable.
Acknowledgements:Theauthorswouldliketoexpresstheiracknowledgementstotheveterinari‐
ansthatcontributedtotheectoparasites’collection,PanagiotisKokkinos,LefterisChalvadakis,Ma‐
riaVafiadou,MariosLiogris,NektariaIoannou,Arsenoglou,FiliposLigdas,ChristinaStrati,Orestis
Dizoglidis,andMarilenaJosephides.
ConflictsofInterest:Theauthorsdeclarenoconflictofinterest.
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... Ectoparasites have an impact on animal wellbeing, cause skin irritation, pruritus, skin lesions (dermatitis and eosinophilic aggregation), and lead to secondary bacterial infections (Abscess formation, pyoderma, and pyemia), anemia due to blood-sucking ectoparasites which may be life-threatening in young and debilitated animals or even toxicosis 'Tick paralysis' which is caused by toxins in tick saliva. In addition, they constitute a zoonotic hazard and transmit pathogens such as bacteria (Rickettsia spp., Borrelia spp.), protozoa (Babesia spp., Hepatozoon spp.) and helminths (Acanthocheilonema reconditum, Dipylidium caninum, and Hymenolepis diminuta) (De La Fuente et al., 2008;Beugnet et al., 2014;Rafiqi et al., 2016 andDiakou et al., 2022). Fleas (genus Ctenocephalides) are the most common ectoparasites of veterinary and public health importance. ...
... Fleas (genus Ctenocephalides) are the most common ectoparasites of veterinary and public health importance. A flea infestation can also cause flea allergic dermatitis (FAD) which is a hypersensitivity reaction to flea saliva (Xhaxhiu et al., 2009;Azrizal et al., 2019 andDiakou et al., 2022). Ticks are the second most common ectoparasite in dogs. ...
Prevalence, clinical presentation, and therapeutic outcome of ectoparasitic infestations in dogs in Egypt
Article
Full-text available
  • Dec 2023
Background: Skin diseases are usually chronic in nature but not life-threatening. They affect the well-being and pose a threat to the general health of the affected animals. Aim: This study aimed to investigate epidemiological, clinical and therapeutic aspects of ectoparasitic infestations in dogs in a number of Egyptian governorates. Methods: Ninety dogs (58 males and 32 females) aged from one month to eleven years from six Egyptian governorates were clinically examined during the years 2022 and 2023. Skin scraping samples were taken from all examined dogs, and deep ear swab specimens from five dogs suspected to have ear mites were obtained and parasitologically examined. Different ectoparasites were classified according to their morphological features. Twenty dogs were treated in four different patterns of administration of local, systemic, and supportive medications. Results: The prevalence of ectoparasite infestation in examined dogs was 64% (58/90). The majority of ectoparasitic infestations (50/58) were single, while the rest (8/58) were mixed. Ninespecies of ectoparasites of fourtaxa were identified: a tick species (Rhipicephalus sanguineus); which had the highest prevalence among isolated ectoparasites from dogs (29%, 26/90), three flea species (Ctenocephalides canis, Ctenocephalides orientis and Ctenocephalides felis) isolated from 18 out of 90 cases (20%), two types of dog chewing louse species (Trichodectes canis and Heterodoxus spiniger) isolated from 2/90 (2.2%) and three mite species: Demodex canis (18/90, 20%), Sarcoptes scabei var. canis (5/90, 6%) and Otodectes cynotis (2/90, 2.2%). The logistic regression analysis of the potential risk factors associated with the prevalence of ectoparasites in dogs revealed that the age, breed, housing environment, habitat, and season were the significant factors affecting the prevalence of ectoparasites (p<0.05 in contrast dog gender did not have a significant effect. Treated dogs showed variations in recovery times and dogs received ancillary treatment showed rapid skin improvement and hair regrowth. Doramectin was effective against ticks and fleas, but Fluralaner was more effective against Demodex mites. Conclusion: The prevalence of ectoparasites in dogs in Egypt could be considered as high and necessitates efforts toward accurate diagnosis, treatment, and control to reduce their impact on animal and public health.
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... Thus, the risk of importation of these agents after returning home is significant (Buczek and Buczek, 2020). Several investigations both in cats and dogs in touristic areas of Europe have shown that this is a realistic scenario that should be considered when owners travel with their pets, and when pets are rehomed in a different country Schäfer et al, 2019a, b;Diakou et al, 2022). ...
Pet travellers' unwanted companions: a parasitic voyage across Europe
Article
  • Jun 2024
In the modern world, country borders do not represent boundaries for the transmission of pathogens. This is also true for parasites that may travel with animals, particularly pets. Dogs and cats are moving across countries more than ever before as a result of adoption, commerce and travels with owners. In addition, climate change supports the establishment of parasites, particularly those transmitted by invertebrate intermediate hosts and vectors, into areas which were previously considered non-enzootic. Therefore, it is important for veterinarians to keep a vigilant eye on such parasitic infections in dogs and cats, be able to recognise compatible signs and feel confident in ordering specific diagnostic tests. This article presents parasites prevalently reported as imported from one European country to another. Furthermore, common clinical manifestations and diagnostic tests are summarised and the preventive measures that should be advised to pet owners are discussed. Raising awareness among veterinary practitioners and pet owners about the risk of parasites spreading in new areas is imperative for controlling and delaying globalisation of important, and potentially lethal, parasitic diseases.
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... In Cyprus, I. gibbosus is associated with vegetated areas (16) and it is not found in dry environments (17). In any case, I. gibbosus is a common species on the island (15,16,18). To explore the herein described geographic restriction of tick paralysis in Akamas, it would be interesting to record the tick fauna composition and the I. gibbosus population dynamics in this area. ...
Tick paralysis induced by Ixodes gibbosus: enigmatic cases in domestic mammals from Cyprus
Article
Full-text available
  • Jun 2024
Tick paralysis is a potentially fatal condition caused by toxins produced and secreted by tick salivary glands. This survey presents clinical and epidemiological observations of tick paralysis cases in domestic animals in Cyprus. Local veterinarians report typical tick paralysis cases occurring in goats, sheep, dogs, and cats. The animals suffering from paralysis are free from other neurological diseases, have blood and biochemical parameters within normal ranges, and recover fast by simply removing the ticks found predominantly on the head and around the neck. Tick paralysis cases occur in a specific geographic area of Cyprus (Akamas peninsula), from September through March, but not every year. Instead, the phenomenon has 2 periodic cycles of occurrence, a 3- and a 7-year cycle. The 2 cycles are differentiated by severity based on the number of affected animals and the resulting losses. As described for other tick-borne diseases, these cyclic patterns may be attributed to external factors, self-oscillations of the disease system, or the combined action of these mechanisms. Ticks collected from a recent paralysis case in a goat were morphologically and molecularly identified as Ixodes gibbosus. Efforts should be made to characterize the specific toxins involved in tick paralysis and to develop a vaccine, which could prevent significant losses of small ruminants, especially in free-ranging farming systems, a prevalent management approach observed in Cyprus and various regions worldwide.
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... birds [4]. Ehlers et al. [5] and Diakou et al. [6] examined flea samples using PCR, and discovered a rickettsia similar to Rickettsia felis. Furthermore, Kumsa et al. [7] found that the chicken flea also transmits Bartonella henselae. ...
Genetic identification of the sticktight flea Echidnophaga gallinacea (Westwood) infesting chickens in the Al-Baha region, Saudi Arabia
Article
Full-text available
  • Feb 2024
Stick-tight fleas, Echidnophaga gallinacea (Westwood), are major ectoparasites of domesticated chickens and can cause serious diseases and even death if left untreated. In the present study the flea samples were collected from three traditional chicken-raising farms in the Al-Baha region. The samples were examined under a stereomicroscope and identified using classification keys. After extracting DNA from the insects, the polymerase chain reaction technique was used to identify the hereditary gene, cytochrome oxidase, present in the insect biopics. The gene was purified, its nucleotide sequence was obtained, and the accession number (OR161051) has been assigned in GenBank. After determining its nucleotide sequence, it was compared with other insects in GenBank, where it was found to be identical (99.82%) to the E. gallinacea isolate from Thailand recorded in GenBank [OQ291364 and MW492259 ( https://www.ncbi.nlm.nih.gov/nucleotide/MW492259.1?report=genbank&log$=nucltop&blast_rank=1&RID=VJR9ZAEE013 )]. The match was 94.33% with Echidnophaga iberica from Spain (KF479239) and 93.97% with Echidnophaga oschanini from China (KU880666). The phylogenetic tree also showed similarities between the Saudi Arabian isolates and other isolates. This genetic study of sticktight fleas is the first of its kind in the Kingdom of Saudi Arabia, therefore will be valuable for assessing the prevalence and geographical distribution of this parasite.
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Dermatological Diseases in Dogs ‒ A Survey in Veterinary Facilities
Article
Full-text available
  • Dec 2023
Skin diseases in dogs are very often the reason for visiting the veterinarian. For the successful management of a dermatological disease, accurate diagnosis, correct setting of the treatment and cooperation of the owner are necessary. This article provides a summary of information related to the diagnosis and treatment of skin diseases in dogs obtained from 50 veterinary facilities in Slovakia. In the monitored veterinary facilities, the most common dermatoses are skin diseases due to immune disorders, followed by bacterial and parasitic skin diseases. For the prevention of external parasites (ticks, fleas), tablets containing fluralaner, sarolaner and afoxolaner are increasingly preferred, even though spot-on preparations, especially those containing fipronil, maintain a constant position in prevention. Among the most commonly used ATBs in the treatment of skin bacterial infections are amoxicillin with clavulanic acid and cephalexin. Currently, the veterinary medicine with the active substance oclacitinib and the medicine containing lokivetmab are coming to the fore in the treatment of allergies in dogs, while veterinarians are trying to limit the use of glucocorticoids.
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Complete bacterial profile and potential pathogens of cat fleas Ctenocephalides felis
Article
  • Apr 2023
  • ACTA TROP
Fleas are important ectoparasites and vectors associated with a wide range of pathogenic diseases, posing threats to public health concerns, especially cat fleas that spread worldwide. Understanding the microbial components is essential due to cat fleas are capable of transmitting pathogens to humans, causing diseases like plague and murine typhus. In the present study, metagenomic next-generation sequencing was applied to obtain the complete microbiota and related functions in the gut of Ctenocephalides felis. A total of 1,870 species was taxonomically recognized including 1,407 bacteria, 365 eukaryotes, 69 viruses, and 29 archaea. Proteobacteria was the dominant phylum among the six samples. Pathogens Rickettsia felis, Acinetobacter baumannii, Coxiella burnetii, and Anaplasma phagocytophilum were taxonomically identified and had high abundances in all samples. The resistance gene MexD was predominant in microbial communities of all cat fleas. We also performed epidemiological surveys of pathogens R. felis, A. baumannii, C. burnetii, and A. phagocytophilum among 165 cat fleas collected from seven provinces in China, while only the DNAs of R. felis (38/165, 23.03%) and C. burnetii (2/165, 1.21%) were obtained. The data provide new insight and understanding of flea intestinal microbiota and provided novel information for preventing and controlling fleas and their transmitted diseases.
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Acaricides Resistance in Ticks: Selection, Diagnosis, Mechanisms, and Mitigation
Article
Full-text available
  • Jul 2022
Ticks are blood-feeding ecto-parasites that have a cosmopolitan distribution in tropical and subtropical regions of the world. Ticks cause economic losses in the form of reduced blood, meat and dairy products, as well as pathogen transmission. Different acaricides such as organochlorines, organophosphates, formamidines (e.g. amitraz), synthetic pyrethroids, macrocyclic lactones, fipronil, and fluazuron are currently used sequentially or simultaneously to control tick infestations. Most acaricide treatments now face increasingly high chances of failure, due to the resistance selection in different tick populations against these drugs. Acaricide resistance in ticks can be developed in different ways, including amino acid substitutions that result in morphological changes in the acaricide target, metabolic detoxification, and reduced acaricide entry through the outer layer of the tick body. The current literature brings a plethora of information regarding the use of different acaricides for tick control, resistance selection, analysis of mutations in target sites, and resistance mitigation. Alternatives such as synergistic use of different acaricides, plant-derived phytochemicals, fungi as biological control agents, and anti-tick vaccines have been recommended to avoid and mitigate acaricide resistance. The purpose of this review was to summarize and discuss different acaricides applied for tick control, their mechanisms of action and resistance selection, genetic polymorphisms in their target molecules, as well as the approaches used for diagnosis and mitigation of acaricide resistance, specifically in Rhipicephalus microplus ticks.
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Culture Isolate of Rickettsia felis from a Tick
Article
Full-text available
Although the cat flea, Ctenocephalides felis, has been identified as the primary vector of Rickettsia felis, additional flea, tick, mite, and louse species have also been associated with this bacterium by molecular means; however, the role of these arthropods in the transmission of R. felis has not been clarified. Here, we succeeded in culture isolation of R. felis from a host-seeking castor bean tick, Ixodes ricinus, the most common tick in Slovakia. The bacterial isolation was performed on XTC-2 cells at 28 °C using the shell-vial technique. An evaluation of the growth properties was performed for both the XTC-2 and Vero cell lines. We observed R. felis in the infected host cells microscopically by Gimenez staining and immunofluorescence assay. The R. felis isolate was purified by gradient ultracentrifugation and visualized by electron microscopy. Fragments of the genes gltA, ompA, ompB, htrA, rpoB, sca4, rffE, and rrs were amplified and compared with the corresponding sequences of the type strain URRWXCal2 and other R. felis culture -isolated strains. We did not detect any nucleotide polymorphisms; however, plasmid pRFδ, characteristic of the standard strain, was absent in our isolate. Herein, we describe the first successful isolation and characterization of a tick-derived R. felis strain “Danube”, obtained from an I. ricinus nymph.
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Current knowledge of vector-borne zoonotic pathogens in Zambia: A clarion call to scaling-up “One Health” research in the wake of emerging and re-emerging infectious diseases
Article
Full-text available
Background Although vector-borne zoonotic diseases are a major public health threat globally, they are usually neglected, especially among resource-constrained countries, including those in sub-Saharan Africa. This scoping review examined the current knowledge and identified research gaps of vector-borne zoonotic pathogens in Zambia. Methods and findings Major scientific databases (Web of Science, PubMed, Scopus, Google Scholar, CABI, Scientific Information Database (SID)) were searched for articles describing vector-borne (mosquitoes, ticks, fleas and tsetse flies) zoonotic pathogens in Zambia. Several mosquito-borne arboviruses have been reported including Yellow fever, Ntaya, Mayaro, Dengue, Zika, West Nile, Chikungunya, Sindbis, and Rift Valley fever viruses. Flea-borne zoonotic pathogens reported include Yersinia pestis and Rickettsia felis . Trypanosoma sp. was the only tsetse fly-borne pathogen identified. Further, tick-borne zoonotic pathogens reported included Crimean-Congo Haemorrhagic fever virus, Rickettsia sp., Anaplasma sp., Ehrlichia sp., Borrelia sp., and Coxiella burnetii . Conclusions This study revealed the presence of many vector-borne zoonotic pathogens circulating in vectors and animals in Zambia. Though reports of human clinical cases were limited, several serological studies provided considerable evidence of zoonotic transmission of vector-borne pathogens in humans. However, the disease burden in humans attributable to vector-borne zoonotic infections could not be ascertained from the available reports and this precludes the formulation of national policies that could help in the control and mitigation of the impact of these diseases in Zambia. Therefore, there is an urgent need to scale-up “One Health” research in emerging and re-emerging infectious diseases to enable the country to prepare for future epidemics, including pandemics.
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Epidemiology, Diagnosis, and Control of Canine Infectious Cyclic Thrombocytopenia and Granulocytic Anaplasmosis: Emerging Diseases of Veterinary and Public Health Significance
Article
Full-text available
  • Dec 2021
This review highlights the diagnostic methods used, the control strategies adopted, and the global epidemiological status of canine cyclic thrombocytopenia and granulocytic anaplasmosis at the animal–human interface. Canine anaplasmosis is an important worldwide disease, mainly caused by Anaplasma platys and A. phagocytophilum with zoonotic implications. A. platys chiefly infects platelets in canids, while A. phagocytophilum is the most common zoonotic pathogen infecting neutrophils of various vertebrate hosts. Diagnosis is based on the identification of clinical signs, the recognition of intracellular inclusions observed by microscopic observation of stained blood smear, and/or methods detecting antibodies or nucleic acids, although DNA sequencing is usually required to confirm the pathogenic strain. Serological cross-reactivity is the main problem in serodiagnosis. Prevalence varies from area to area depending on tick exposure. Tetracyclines are significant drugs for human and animal anaplasmosis. No universal vaccine is yet available that protects against diverse geographic strains. The control of canine anaplasmosis therefore relies on the detection of vectors/reservoirs, control of tick vectors, and prevention of iatrogenic/mechanical transmission. The control strategies for human anaplasmosis include reducing high-risk tick contact activities (such as gardening and hiking), careful blood transfusion, by passing immunosuppression, recognizing, and control of reservoirs/vectors.
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Mediterranean Spotted Fever: Current Knowledge and Recent Advances
Article
Full-text available
  • Sep 2021
Mediterranean spotted fever (MSF) is an emerging tick-borne rickettsiosis of the spotted fever group (SFG), endemic in the Mediterranean basin. By virtue of technological innovations in molecular genetics, it has been determined that the causative agent of MSF is Rickettsia conorii subspecies conorii. The arthropod vector of this bacterium is the brown dog tick Rhipicephalus sanguineus. The true nature of the reservoir of R. conorii conorii has not been completely deciphered yet, although many authors theorize that the canine population, other mammals, and the ticks themselves could potentially contribute as reservoirs. Typical symptoms of MSF include fever, maculopapular rash, and a characteristic eschar (“tache noire”). Atypical clinical features and severe multi-organ complications may also be present. All of these manifestations arise from the disseminated infection of the endothelium by R. conorii conorii. Several methods exist for the diagnosis of MSF. Serological tests are widely used and molecular techniques have become increasingly available. Doxycycline remains the treatment of choice, while preventive measures are focused on modification of human behavior and vector control strategies. The purpose of this review is to summarize the current knowledge on the epidemiology, pathogenesis, clinical features, diagnosis, and treatment of MSF.
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Exposure to Major Vector-Borne Diseases in Dogs Subjected to Different Preventative Regimens in Endemic Areas of Italy
Article
Full-text available
  • Apr 2021
Vector-borne diseases (VBDs) are globally widespread arthropod-transmitted diseases with a significant impact on animal and human health. Many drivers have recently spurred the geographic spread of VBDs in dogs. This study has evaluated the exposure to most important VBDs in dogs under different preventative treatments in different regions of Italy, i.e., Veneto, Friuli Venezia-Giulia, Umbria, Giglio Island (Tuscany), Abruzzo and Latium. Serological analyses were performed to detect antibodies against Leishmania infantum, Babesia canis, Anaplasma phagocytophilum/Anaplasma platys, Ehrlichia canis/Ehrlichia ewingii, Borrelia burgdorferi, Rickettsia conorii and the circulating antigen of Dirofilaria immitis. Dogs were categorized according to the treatment schedule usually received, and the association between seropositivity and possible risk factors was statistically evaluated. Overall, 124/242 (51.2%) dogs tested positive for at least one pathogen, while 34 (14.0%) were exposed to two or more pathogens. The most detected seropositivity was against R. conorii, followed by Anaplasma spp., L. infantum, B. canis, and the other pathogens under study. Significant statistical associations were found according to geographical provenance, history of tick infestation, lifestyle and inadequate prophylactic treatments. Random/irregular treatments have been identified as a clear risk factor. These results show that adequate prophylactic treatment protocols are overlooked by dog owners, despite the availability of several effective products, with possible implications in veterinary medicine and on public health.
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Recent Advancements in the Control of Cat Fleas
Article
Full-text available
  • Sep 2020
With the advent of imidacloprid and fipronil spot-on treatments and the oral ingestion of lufenuron, the strategies and methods to control cat fleas dramatically changed during the last 25 years. New innovations and new chemistries have highlighted this progress. Control strategies are no longer based on the tripartite approach of treating the pet, the indoor environment, and outdoors. The ability of modern therapies to break the cat flea life cycle and prevent reproduction has allowed for the stand-alone treatments that are applied or given to the pet. In doing so, we have not only controlled the cat flea, but we have prevented or reduced the impact of many of the diseases associated with ectoparasites and endoparasites of cats and dogs. This review provides an update of newer and non-conventional approaches to control cat fleas.
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Canine and Feline Parasitology: Analogies, Differences, and Relevance for Human Health
Article
  • Aug 2021
  • CLIN MICROBIOL REV
Cats and dogs are treated as family members by most pet owners. Therefore, a high quality of veterinary care and preventive medicine is imperative for animal health and welfare and for the protection of humans from zoonotic pathogens. There is a general perception of cats being treated as “small dogs,” especially in the field of clinical parasitology. As a result, several important differences between the two animal species are not taken into proper consideration and are often overlooked. Dogs and cats are profoundly different under evolutionary, biological, ethological, behavioral, and immunological standpoints. These differences impact clinical features, diagnosis, and control of canine and feline parasites and transmission risk for humans. This review outlines the most common parasitoses and vector-borne diseases of dogs and cats, with a focus on major convergences and divergences, and discusses parasites that have (i) evolved based on different preys for dogs and cats, (ii) adapted due to different immunological or behavioral animal profiles, and (iii) developed more similarities than differences in canine and feline infections and associated diseases. Differences, similarities, and peculiarities of canine and feline parasitology are herein reviewed in three macrosections: (i) carnivorism, vegetarianism, anatomy, genetics, and parasites, (ii) evolutionary adaptation of nematodes, including veterinary reconsideration and zoonotic importance, and (iii) behavior and immune system driving ectoparasites and transmitted diseases. Emphasis is given to provide further steps toward a more accurate evaluation of canine and feline parasitology in a changing world in terms of public health relevance and One Health approach.
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Vaccine approaches applied to controlling dog ticks
Article
  • Jan 2021
Ticks are considered the most important vectors in veterinary medicine with a profound impact on animal health worldwide, as well as being key vectors of diseases affecting household pets. The leading strategy applied to dog tick control is the continued use of acaricides. However, this approach is not sustainable due to surging tick resistance, growing public concern over pesticide residues in food and in the environment, and the rising costs associated with their development. In contrast, tick vaccines are a cost-effective and environmentally friendly alternative against tick-borne diseases by controlling vector infestations and reducing pathogen transmission. These premises have encouraged researchers to develop an effective vaccine against ticks, with several proteins having been characterized and used in native, synthetic, and recombinant forms as antigens in immunizations. The growing interaction between domestic pets and people underscores the importance of developing new tick control measures that require effective screening platforms applied to vaccine development. However, as reviewed in this paper, very little progress has been made in controlling ectoparasite infestations in pets using the vaccine approach. The control of tick infestations and pathogen transmission could be obtained through immunization programs aimed at reducing the tick population and interfering in the pathogenic transmission that affects human and animal health on a global scale.
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