A TAXONOMICSTUDY
or
THEHAEM OPR OTEW AE~,\ P ICOMP Lr. XA:HAt:MOSPORINA)OFTHE AVIANFA:'.IIUESFR ING ILLI DAE, EM UERIZ II>AE,PARI; U DAF...THRAUP IDAEANDICfERIDAE
by
Jenn iferRuthBurryCaines.R.Sc.(Hons.)
Atbcsts submittedto the Schoolof GradunteSiudiesinpartla!fulfilrnenr
ofIhe require ments forthedegree of Master ofSclcncc
Departmen torBiology Men' orinl Un iversityofNewfoundland
July,1991
St.John's NN foundlan d
1+1
NatiOflalUbrary ofcanadaBibliol~uenalional p.
eoceeee
Canadian Thesesseoee sevce des eeses canadHwlCs
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Canada
ABSTRACT
Thehaemoproteids(Hae mosportnc :Hncmoprotcfdnc )oftheavianfmnilies Fringillidae,Emberizidae,Purulidne,Thrauploaeand Icte rid:IC S('1IS1lEdwar ds(!')s6) werereviewed,andeightspecies determined\0he vulid.allofthe basich:lltcridi:l!
or microhulteridiulfor m asdefinedbyBennett andPeirce(!9RH ).1/{/(·lIlf1pm/('u.\·
jrillgillae(Fringillidae). H. mazzai(Emhe rizidae) , H. chlods(Emherizid:lc)undII.
quiscalus(Icteddae )wereredescribedfrom hapnntmy peor ncobupunroryp cmate rial fromawiderrungeof h0515and/o rlocationsthunin theorigin:!! dcscriptlrms.
Haemoproteus tosiae, H. oconthis andH. embaizawere declarednominanuda. II.
8lofJulosu.~, H. lIIucrop;8melltulus,II.sai ni;lind H. lar kuk()I'.\-kyiwere decla red synonymsofH.chlotis,andH.hedymeliswasdeclar eda synonym ofN. muezoi, 11\c newhuemoproteld species Haemoproteus coamevi (Emheriz idae ), II. Ilt/ntli (Parulidae) ,H. coereba (Parulidae ),andH.thmupi(Thraupidae )weredescribed.
All hap antotype,neohapantotype andparahapanto typc mutcrmlWHSdepositedinthe collectionof theInternational Refe renceCentreforAvia n Hucmarozou.Dcpnrtmcm of Biology,MemorialUniversityof Newfoundland .
Iii ACK NOWLEDGEMENTS
I wishIIIthank my supervisor Dr. GordonBennettfor hiscontinuedsupportand gu idanceoverthe years.
Ialsowishtothanktheother membe rsofmy supe rvisorycommittee,Dr.William Thre lfallund Dr.Ja keRicefor their helpful commentsalongtheway.
ThankstoG.valktnncs for allowing meto examinebloodsmear material from Lithu ania.
Tha nksalsogoesto MadonnaBisho p forherever lastingpatience and support.
Thanks toDebbieSquires-Parsons forher encourage me nt andsupportwhen everyone else hadgone.
Tha nkstoCuria Wc odworth-Lynusfor all the pertinent andpersistentquestions.
Thanksto myhusba ndDoug, for bearing withme duringthelong,longstretch.
Thanksto my parents Margar etand HubertBurry, for believingthat "when you put your mind 10 it.
TA8LEOFCONTE NTS
ABS1l{ACf •..••. • •.••..•. •... •..•.•..• •. • • • • • •. . • .... • ...ii ACKNOW LE DGE MENTS .... . ... . ... .•... ..•... . . ..•.. .•.• •iii LIST OFTABL ES••...•.••.• •.•...•...• •.. •....••. ..
LIST OF PLATES .
INTR O DUCTION '
MATER IALSANDMETHOD S " ..
.. ...vi
..vii ..I
Preparationofbloodsmears .
Taxon omicstudyofbuemoproteld s .
. ..4
RESULTSANDDISCUSSiON... .•...
TAXONOMY
••••••• •••X
Historical •••.••...•...••• .•.••.•. •. . ...•• .•• . .•.•••I'!
Life cycle .• •.... . .. ... ... .... .•.. . .. ... . .. ...lJ
Morphology .•• . . ... .... ... ... . .... . . . II Taxonomicstud)' ofHaemopro tclI,f•.•••••.••• • .••• •12
••••• • • • ••1(1 TAX ONOMICREVIEW..•.•..• ...• ... . .•. . . .. .... .. ..HI
FRINGI LLIDA E
.....67 27
. 27
.. 34
. 40
..50 .•.50
... ...59 ... ..63 Emberizinae..
Cardinulinue..
Carduelinae..
Purulinue Coerebinne PARULID AE ..
EMBERIZIDAE...
THRAUPIDAE ....
ICfERIDAE..., ... .. . . ... . ... . ... . .. ...
SUMMARy ,... . . ... ..72
LITERATURE CITED . . .. ...•.. •... 76 APPENDIXA... .... . . ... . ... 8J
Tabl e
LIST Of TABLES
"a I-'\'
Classificationsofavia n hostsreviewedin the curre ntstudy...••. . .•••. .•... . ...•.
Morphometricpummctcrsofthe haernopmt ehls nfthe
Fringillidueand Emberizidae .•.IX
Morp hometri c parameters of thehacrnopro rcldsofthe
Parulidae,Thraupida eand Icteridae.•. ... . . . . .. .. ... ....•. . .51
HaCllloprofCUSspecies of theFringillirJae,Embena id ae
Parulidae,ThraupidaeandIcte ridae.•. ... .. ... ...75
I'la le
A
vii LISTOF PLATES
FollowingPa ge
...•...•.14
Jtaonoproteus [ril/gil/ae.•. . .•..•.. .• ..•.•. . .• •. . . ...17
Haemoproteuscoameyin. sp. ..•.,... . ... . .... . ...28
Haemoproteus nuazai .. ... . . .. .. ..•... .. . ... . 35
/ taemcproteus chkxis ..•...•... • . .. . •. .•... •... •... •.•.42
Haemoproteusparulin. sp. ... .•.
Haemoproteuscoereba n. sp.. ..
. 51
. •..60
Haemoproteustllrt/up; n.sp. . •. . . ..• . . ....•... .... ...64
Haemoproteusquiscaius.. ....•.... .... .•• ... ... . ... .• •. ..68
INTHODUCfIO N
The avia nIamiliesFringillidac.Emhcril illac.Panllitla~.Thr:lIIJlid;lCarul lcteridae$('IlJUEdwards (19M) area.,~mhlagesof nine-primaricd passerine hinls.
speciesof whicharcfou ndworld wide, occupyinga diverse rangeof niche!' (l'eten.
1970.Campbelland Lack 1985). Unde r vartous~ystcm~IIfclassificutiouthcyinclude thetruefinches,carduelincfinc hes,Hawaiianho ncyerccpcrs.eonebills,l'anl inals, canaries.buntings,NorthAme rican wood warblers.bnnnnaquits, tanagersnml grackles and arcthoughttorepresent someofthemoste\'ohrtinnarilyndvnuccd passe rinespecies (Cam phellandLack lQRS), Newtmerprct uuo nsofunatomiculdmu andthe developm entofblocbemlcaltecb ntquesincludi ng ctectropborcsrsundDNA hyb ridization(Sil'lley and Ahlqu ist 1984) havercsuhcdin major cha nges in,lVi;lll taxonomyin recentyears, particularlyin location uf gcncruwithinf"milygrnll pillg.~, In1983,the AmericanOrnithol ogists'Un io n (A,O,U.)revise d their check-listHfthc hirdsofNorthAmerica;in parti cula r,the ch,'l'k·!ist changedthe orde randcurucnt cfn'anyfamiliesand red uced severalfamili..·..tu sub-Iamiliulstatus.Clements(\117X) alsoreorg anizedanearlier(1974 ) ed itionuf his worldwidecheck-lis t,andEd~lJd!' (1986)pub lished anewand differe nt editinn ofthe 8lrdJnft"!! World. alteringthe fa mily statusofsevemlgroups, Edward'scheck-list was rhn..en fur the currentstudy asthemost recentandmostcom prehensivetrea tmen t
or
pilsserifnrJl1 sysrcuuuics.Theresultsof thesecha nging classificationsbuvcunimpac t on taxon mnic studiesof theavian bloodparasitesof thegenuslIur:lllop m/('/H. BiHI hlJ<,I·family.
and/or subfamily specificity,successfullydemonstrated in cross-tran smissio nstudies
1
.1 foreight hostbird families,isone ofthe maintaxonomic criteriaof thisgroupof upicomplcxun protozoan s(BennettandPeircelQR8,Atkinson 1986).In most of the avianfamiliesstudied to date, thishost-family (su bfamily) specificity has beenused in conjunction with the morphological featuresof the erythrocyte-inhahirin g gamctocytesin thedescriptionof new parasitespecies. The gameto cyresure productsofthe asexualdevelopmental stageoccurringin theavian host,and are the primary diagnosticstage.Gametocytemorphologycanhevariable withina particular specks bothat thesa me anddifferentgeographicallocations, among host species withina family, andindeedwithina particular individualOWltime(BennettaridCampbel l 1972, 1973. Bennettettil.1985, Bennettel af.1986, Bennettand Pe.rce1988).Therefore, inthe absence (If cross-transmissionstudies, speciesdiffereutiatirmof hacrnoprmeid parasiteswhich aremor phological lysimilar isstilllargely based onthe familyof the hirdhost(Bennettand Peirce1988).
Atota l of 12 speciesof 'haheridialtype'Haemoproteus(Bennett and Peirce 198H)have heen namedor describedfrom membersofthe Fringilltduc.Emherizidae, Pnrulidae, Thruupidaeand lcteridne,some onthe basis of the 'one-host- one parasite' concept,others onthe basis of morphology.Eightof these species were originallydescribedbefore guidelinesfor defining a species were published (Helmy Mohammed195M,BennettandCam pbell1972 etseq. ) Threespecies,H.ocanthis, II.embcrisaandfI.Iosiaehavesimplybeen named in publicationbut ther e are no
species descript ions. Mosthaemoproteids Inthebirdfamiliesconsideredin the curren tstudy havebeenidentifieda~Haemoproteus sp..H. [ril/gil/m'.1-1.orizivome (Ben nettetat.1982,Peirce 19R4). Inpractice.the term'1/.!rillgillm:/orizil'fmw complex'(Peir ce1983) hasbee n used to identifythehaemoprote lds foundIn many hostfamilies in theabsence of detailedstudieson theseparasites.Thedlstrllnuiou ofHaemoproteus oruivomehas recently bee nrestricted til theaviansuhf;unily Poe philinae of the Estrtldidae(Benne tt andPeirce191)I),Haemo proteus[rillgil/ae isthe only species describedfrommembers of the Friugillidae.Within the family Embcr izldae,sevenspecies havebeen describedfrom the Cunluclinuc:H.{{e(/JIII/ix, H.kuioe,H.cntons,H.glohuiosus,H.macroptgmenuuus,H.sainiandII.tart akovskyi;
twospecies from the Cardtnatlnac:H.nuuzaiandfl.hedymclis;lind one species fromthe Brnbc rizinae,H.emberizu,OnlyH, qulsculushas beendescribedfromthe Icte ridae. Nohaemo proteid specieshave beendescribed fromtheThra upidae or Par ulidae (subfamilies Parulinae andCoereblnue).
The collectionoftheIntern ationalRefer e nce CentreforAvianl lucmutozou conta inssamplesof blood smears taken from ma ny of thesebirdspeciesacross their distributiona l range, and in some casesfromthesame individualoveraperiod of time.This materialprovides the baststo evaluatethe huemoprotcldsfromthese familiesand determinewhich ofthe12 curre ntly describedspeciesurc validand whethe r additional speciesoccur.
MATE IUA LSANI) METHODS
The blood smea rmaterialfor thistaxonomic study wascollected by con tributorsfromaroundtheworld,including the author,and deposit ed in the collectionoftheInte rnational ReferenceCentrefor AvianHacmatoaoa(fRe AH ), Department of Biology,MemorialUniversityof Newfoundland . Allsam ples were from wild-caughtbirdsand wereofnaturalinfections.Somebirdswereretrapped and providedsamples ontwo or moreoccasions. Thedatasetis extens ive; over 1,200 blood smearscontaininghuemoproteldswereselectedfro m thecollectionand examined.The hostspecies andgeographiclocationsof selectedspecimensaregiven in AppendixA. Table1 liststhe classiflcatlonsof thehost families under considerationaccordingto Edwards (1986), Clements (1978)andthe A.O.U.(1983) forcomparativepurposes. Parahapantotype materialwas available for two J-/acm opmtellsspecies . Alltype mate ria l wasdepositedin thecollectionof the IRCAB.
Prep aration of bloodsmears
Bloodsmears were taken from livenettedbirdsby prickingthe brachialvein III obtaina few drops of peripheralbloodandmakingathin smearfollowing the protocolsof Dennett(1970).Some materia lfromblood of deadbirds was examined bUIwasof littletaxonomic use. Theslides wereair-dried,fixed in 100% metha nol orethan ol,and stained with a varietyof stains (Glemse's,Wright's, Hasting's). Some
TableI ga 5_~m~ationsoravian host s rertewed Inthe curre ntSllidy.
Edward s1986
FRINGILU D AE Fringillinae
EMBERIZIDAE Carduelinae Cardin alinae Embe rizinae
PAR ULIO AE Paru linae Coerebinae Conlrost rtnae
THRAUPIDAE
ICfER ID AE lcierfnae Dollconychlnae
Clements 1978
FRINGlLLlOAE (cont ains speciesfro m
E d wa r d s '
Emberizidae)
PARUUDAE
CQE RE BIDAE
THRAUPIDAE
ICfERJD A E
A.O.V.1983
FRINGILLIDAE Fringil1inac Carducliuac
EMOERIZIDA E
Cnrdinalinnc Ember izinac
Purulinac
Cocrcbinac
Tlv uupinac
lctcrinuc
materialwas stainedinthecountryof origin, but most werestainedat the IRCAH withGlemsa's stain (buffered to pH7.2)for 45 minutesandwashedin slightly acidic (pHIi.S)lapwater.Initially, all smears were scree nedby the contribu tororIRCAH staff, and where possibleparasiteswereidentified to genus. Forthis study.slides were examined under 40x and lOOx oil immersionobjectiveson a Zeiss II Photcmlcroscope, for thelength of time necessary to identify the haemoproteids (up to120minu tes per slide).
Taxonomic study or haemcprotelds
Samplesof uninfected erythrocytes and parasites within infectederythrocytes of selectedbird specieswith in eachfamily (40individualho st birds) weredrawnand measuredwithaZeissMOP-3 DigitalAnalyzer. Photomicrographswereobtained using a Zeiss IIPhotomicrosco pe. Line drawingswere made fromWIIlC'r alucida drawingsonaZeiss RA microscopeusing a 20x objective.More macrogametocytes than microgarnctocytes were drawnbecause of theirrelativefreque ncyand valuein species diagnosis(Bennettand Campbell1972). Gametocyte length, width and area, gametocytenucleus length,widthandarea,erythrocyte length,widthandarea,and crymrocytenucleuslength , widthandareawere measuredand various ratios derived according10the protoco lsofHelmy Mohamme d(1958),Bennett and Campbell (1972)and Forresteret al.(1977). Someofthese ratioswere usedto indicate the relativehypertroph yor atrop hyof erythrocytescaused by theparasite and thesize ofthepara site relativetotheinfected cell. Inaddition,the nuclear displace ment
ratio (NOR)asdefinedbyBennettand Cam pbell(1972). was used 10 indicate 1111:
degreeof latera ldisplacementof theerythrocy te nucleuscaused hy the gametocyte. The number,arrangement.sizeandshapeofpigmentgranules.oftenchamcterisuc of haemoproteidspecies werealso recorded.as wasthe presenceofvolutin granules (vonBra nd1966,White andBenn ett197R).Where possible,gametocyte sexrillius werede termined, andqualitativecharacterssuch as pcrasheoutline andnuclear positionwere recorde d.Informatinn onthe age andsex of thehostbird,.ISwellas the dateof captur ewerenoted to assesswhat effectthesemighthavenn the parasite's morpho logy.
RESULTSAND I>ISCU SSION TAXONOMY
Historical
Kruse(1890)first erectedthegenusHaemoproteusand describedH.cofumbae in the pigeonColumbalivia. The parasiteoccurring in thepigeons (Columbidae)was studiedbySerge ntandSergent(1906,1907),Aragao(1908),andAdie(1915)who confirmed the vector as alousefly, LYl1cllin.maura(Hippoboscidae ). For manyyears following,itwas assumedthathtppoboscklswere thevectors ofall haemoproteids.
Newspecies were named or describedfromseveral avian families,oftenbased ona'o nehost species -one parasitespecies' conceptratherthanstudiesof morphological differences.This resultedinaproliferation (almost200)of species namesand/ordescription s, many of which are inadequatebymode rn standards (Dennettet01.1982).
As life-cycles of vario us species became known and experimen talcross- transmissions werecarried out, the 'one hostspecies • oneparasite species' philosophywas serio uslyquestioned.Experimenta lwork (summar ized by Ben nett er al.1985)demo nstrated thatspeciesofHaemoproteus weretransmittedprimar ily by members of theCeralopogo nidae(sevenof the11speciesfor whichthelife cycle and/ or vectors areknown)and thatparasites were host-family orhost-subfa mily Si1:::.:ific (Bennettetaf.1982,Atkinso n1986),butnotspecies specific.Thishost- fumily(subfamily) specificityisused inconjunctionwiththe gametocyte's morphology intheide ntificat ion ofHaemoproteus spp.until exper imenta lworkcan co nfirm the
validity of suchspecies (Be nnettand Peirce1988).
Levine(1980) places thefamilyHaemop m tcldue,conminlngthegenu~
HaemoproteusKruse,1890in theSubo rderHaem ospod na Dnnilewsky.IKR5orthe PhylumAplcomplexaLevine,1980,Thegenusincludesuncut IDspeciesorpnrusitic protozoaof birds(Ben nettand Peirce1988).De nnett andPeirce(Il)H8)discussed the morphologi calformsof the avianhacmoprot cids andpresentedacheck-listof 113 species,manyof whichhaveheenrecently rcdcscribe dhy Bennettandhls colleagues. Haemoproteusspecies haveaheteroxenouslifecycle,undergoing schizogony inthe vertebratehost and sporogony in theinve rtebrateho~twhichacts as the vectorfor the tra nsmission ofthe purushe from onehirdItJanother.For most species ofHaemoproteus,there is little evidenceofmorbidityor mortalityassociated withinfection (Bennettet uf.1982,1988). Howeve r, pathogenicityh"shccn demonstratedinH.1Ile/eagridisof turkeys(Atkinson19116) anddoves (Bennett pcrs.
comm.),andhasrecentlybeen implicate d inHaemoproteusgaffi,schlzontsofwhich mayhave been miside ntifiedasAnhro cystisbyLevineelal.(1970) and whichcause extremelesionsin heartmuscle(Bennett, pers. ccmm.).
Llfecycle
The life cycles ofthose speciesstudiedhave a schlaogcnlcstage in thelungs, heart,kidneys,liver andotherinte rnalorgansofthe birdhost(Benne ttel at.!1J65, Desser and Bennett in press). Sporozoites,when inocula tedintothe peripher al blood by thevector,enter these internal organs,and eachdevelopsintoaschizont.
10 Schizogonicdevelopment may take5-17days aftersporozoite -inoculation. Each schizontproduces a number ofrnc rozoites. Initially, these merozci tescontinuethe tissue stage(eachprodu cing anotherschizont, recurrent cycles producing'flrst-' and/ or 'second-generation' schizonts). Eventually,some merozoites enter erythrocytes, and each forms either a macroga metocyte (female) mrcrogame rocyre (male). Maturationof a merozoite usuallyrequires 4-6 days (DesserandBennett ,in press). The prepatentperiodaverages 14days formost species. Mucrogametocytes and microgametocytes arepicked up viaa blood meal by the vector to begin the sporogonic stage.The vectors foronly 10 species are known,eightofwhich arebitingmidges of the genusCuIicoides(Ceratopogonidae).
Louseflies oftheHippoboscidae arethoughtto be vectors ofthe other species.
However. for these species. there is now circumstan tialevidencewhich suggeststhat Culicoidesarealsoinvolved andthathippoboscids areunusua lvectors (Desserand Benn ettinpress).
After the gametocyte-infected blood isingested.the macrogamet esareformed insidethe macrogametocyte and the mlcrogametes arerap idly released inside the midgut by a process termedexflagellation.The rnacrogamereis fertilizedby the microgametetoform a zygotewhich developsinto a ookinete. The ookinete penetra tes the midgutepithe lium 10the basementmembra neofthe gutand encysts the re toform anoocyst. Sporoecltesareformed(usuallyabout 16) within the oocyst and released throughoutthebody ofthe vector.Someaccumula tein thesalivary glandsandarctran smittedto another host viaabloodmeal,continuingthecycle
II whenthe sporozoitesenterthe various organs fromthe peripheralblood Depending upon thespecies ofHaemoproteusand theambient temperatureat whichthevectors were held. the sporogoniccycle may take a minimumof7- 12days (Khanand Fallis
\969).
Haemoproteus spp.infectionsresultin a pro longedpumsitacmiu '1Il~1in temperateregions,intensityincreasesbeforedecreasingto;Ilowerchroniclevel.
Often this stage revealsonly a few circulatinggametocytcs.Cycling with theonset of the reproductivecycle inthe hostbirds isa phenomenon{Ifrelapsein which presumablyhormonal changes andstress cause anincreasein intensity
or
infection with an increase in the numbersofgametocytes (Bennettand Cameron11J74).Morphology
Onlythe gametocytesofHaemoprotC'us are seen in the peripheralblood of the avianhost. Macrogametocyte s(female) andmicrogamcrocytcs(male) occur in erythrocytes,with fewspecieshaving distinct ivefea tu res.Bennettand Peirce(19KX) classifiedalldescribedspecies asoneof fivemorphologicalforms.Gnmctocytcs of mostspecies form either a small orlarge yokeor 'halteridi um'around the erythrocyte nucleus(mlcro-halt erfdia lami halteridial forms),whileothers completely encircleit(circumnuclear ). There is also a broad cylindricalformwhich nften enucleates thehost cell (rhabdosomal),and ararer disc-shapedfor m (discosomal).
For most speciesthe gametocyteisthe onlyknown stage,Theidentificutio n and rea ring of bothvectorsandhost birds present treme ndouschallengesforlife cycle
12 studies (Atkinson1986) and classification forHaemoproteusspecies has been based primarilyonhosttaxonomy (Bennett and Peirce 1982). For many speciesit was recognizedthut future studies might dete rminetha tthey werenot distinct biological species, and would thenfall intosynonymywithpreviously describedspecies.
The morphologica lcharacteristics of the gametocytes thathave been used to diffe rentiatespecies of the same basic formare: gametocyteoutline, size and shape, hypert ro phy,nucleardisplacement ratio (NOR), the number,size andplacementof pigmentgranules,presence ofvolutin granules.andothers(HelmyMohammed 1958, Bennett and Campbell1972, Bennettetc.!.1984, 1985. 1988, Greine ret ol: 1977).
Variability in mensu rulcharacters has been viewedas natural variationdueto host, geographiclocation orseasone rdifferences (Bennettet at.1984,Be nne ttetal. 1987).
Taxono micst udyofHaem oprot ew
Although theuse of dried bloodsmearsis a valuab lediagnostic tool for haemoproteid study, the rearelimitatio nswhich restrictquantitativestudyof the characterscommonlyused in taxonomicanalysis. The materialis subject to artifacts of preparation (Bennettand Campbell 1975), which oftencause variabilitythat dominatessuch analyses. Severalfactors can affect the appearance ofthe gametocyt e.
One of the difficulties intaxonomic studyof blood parasitesis withblood smearrnuteria ltake nfromdead birds (Be nnettandPe irce 1988). Post mortem changes include the gnmetocytesrounding up and/or po pping outof theinfected
13 erythrocyte. This actually simulates thefirst stages of garnctogencsls. which normally occurs in the gut of thevectorafteringestion.At leasttwo species of 'globu larIYP~' Haemoproteushave been describedfromfJO.1'lI1IO,"ICIIImaterial undhave sincebccu synonymized with hulteridialspecies upon studies of suitnhle material (While ami Bennett1979.Bishop and Bennett1989).Material fromdeud birdswas examined in the currentstudy,and rounding up and poppingout wasobservedundattributed to theseposl mortem changes (PlateA,Figure 1).
Blood which is slowdrying, eithe rbecauseiIwas very thicklysmeared[ideally smears should be onelayer of cells), orduetilextremely humidconditions,can ulso cause simila rartifacts.Parasitesnear smear peripheriesmay thentc ndto he normal whilethose near the centre may he roundedas withpastnlOnC'1IIhlood{Bennett, Gabaldo nand Ulloa 1982).Delayed orimproper fixingand stainingalso can have effects. Ofte n theparasite 's nuclearmaterial condenses andshows deeply stained spots. Observations of such characters,alongwiththose of thenor mal uninfcctcd erythrocyteswereusefulin determining if slow-dryingwas a factorintheappcuruncc of the parasites.
Excess mechanicalpressureonthe microscope slidewhen the smear is mudc distorts and somet imes ruptu res cells, Whenuninfected erythrocytesarcdistorted in this way. the exact shape andsize oftheparasite s are likelytohavebeen similarly altered, Even withoutexcess pressure ,attheouteredges ofthehloodsmear the parasitesare oftenbroaderand largerthan those inmore central areas.They al!;{)
Figur e1.
Figur e2.
Figure3.
Figur e4.
Figu re5.
Figu re6,
PLATE A
Microgametocyte ofH,cntons,initiat ingexflagel lation Macrogamet ocyteofH.quiscalusnearsmearperiphery GametocytesofH.coameyi
Macr ogameto cytesofH.c1Jlorisdisplacingnucleus Macrogarneto cytes ofH,paruli
MacrogametocyteofH. quiscalus
-' - - ... - -
- .,
l
.,
~, -
... ..
20p m15 appea r to camemore displacementof the erythrocytic nucleus (Plate A.Figure 2).
Some timesthe observationof 'fat'paras ites result from mecha nical factors,asthose Cor ms arc usually seenonthesmearmargins, and sma ller halter idial gametocyt esin themiddl e ofthe smear, and caremustbetakentodisti nguish artifactsof the slide preparatio nwithtruenuclear displacement. Comparisonwith normal uninfected erythrocy tesnear the infected cells canoften help in thisdeterminatio n. Inadditio n tophysical factorsaffectingthe appearanceof parasites,otherinfluencesinclud e the uge ofthe hostbird. whetheritis a chronic,relapse, or a newinfect io n,andthe developmentalstage andinte nsityof theinfecti onitself.Asthe material used inthis study was obtainedfrom wild caughtnaturally-intectedbirds,itis difficulttofollow an infe ctionthroug h fromimmatureto maturestages.Reinfectionmaybe expected 10 occur throughout themaintransmissionseason,andit wasthereforenot possible 10 separate gameto cytesof singleoriginaswo uld be possible forbirds incaptivity.
Ther e was sufficientmaterial for only afewho st birdspeciesforeffectiveevaluation of some of thesefactor s,and these observa tionsareincludedinthe respe ctivespecies descrip tions.
II,
TAXONOMICREVIEW
fRINGILLIDAE
Haemoproteus [ringillaeLabM189~
TypeHost:Fringilla cocfebsLinnaeus [Chaffinch]
Type Locality:BramleyHants, England
Immalureg3me~(PlateI,Fig 1):You nge st formsseeninitiate growth ncarthe middleof erythrocyte,rarely nearpoles.
Macrogametocyte(Pla te 1, Figures2,4-5 ; Table2):Halte ridialparasite ofmed ium
10 largesize, occupying60·70%or the erythrocyte-parasite complex,not causinghost- cell hyper trop hy; cytop lasmfinelygranular,stainingmoderat e-bluewithOlcmsn's
stain;marginofparas iteentireandusuallyappr esstngthe hostcellnucleus,hut no\
always the centralper ip heryofthehost cell; ends ofpar asiteusuall yrou nded,net always touching poles of erythrocyte, nor withsymmetricalcurvesaround the
erythrocytenucleus;hostcellnucleus notusuallydisplacedlaterally(NO R",0.96);
pigme ntgranul esroundtooval,often largeand rod-shaped(uptoll<!m in diamet e r), yellow-brown,averaging 13 {rangi ng8-25) , usu allyrandomly distributed thro ugh o ut thecytoplasm; volutinrarely seen; pa rasitenucleuscompact,round to elongat e oval,staining pale pinkwithGiemsu's stai n,terminaltosub-terminal inposition,
PLATEI
Haemoproteus fringillae
Figure1. immature gametocyte
Figure 2. immaturemacrogametocyte, with'dip' Figure3. mlcrogumeto cyte
Figure 4. macrogametocyte,la rgepigmentgranule!'.
Figure 5. rnacroguracrocyte
t ojrm
18 TABLE2. Morphome tric par amete rs ofthe haemo prot eids of theFringilli daeand Emberlaldae.
H.fringiflue H.coameyi H.nuazai H.chk ris n.sp.
Unin fectcderythrocyte N=45 N
=
135 N=111 N=
70length 12.6 11.9 11.6 11.6
(.87)" (.81) (.85) (.75)
width 6.2 6.2 6.5 6.2
(.32) (.57) (,46) (.55)
62.5 59.2 59.8 58.5
(4,4) (7,4) (6.2) (7.6)
Uninfected erythrocyte N=45 N=135 N;. 111 N '"70 nucleus
length 5.8 5.5 5.0 5.5
(.45) (.50) (.59) (,40)
width 2.5 2,4 2.2 2.5
(.24) (.27) (.29) (.25)
12.0 11.0 9.1 11.3
(1,4) (1.8) (1.6) (1.6)
Erythrocyte infected
bymacrogametocyte N==75 N=150 N =159 N
=
86length 13.0 12.3 12.8 12.7
(.91) (.94) (.95) (.63)
width 6.1 6.2 6,4 6.5
(,42) (.70) (.51) (.65)
64.8 63,4 66.7 67.2
(6.5) (8.7) (6.3) (6.6)
I"
TABLE2 (eont'd), Morphometricparametersofthe hacmoprotclds ofthe Frlngillidae and Emberizidae.
H.Jrillgiflue H.coameyiJf.lI1azw; il-chlcri s n.sp.
Infected erythrocyte N<75 N "" 150 N '" 159 N =XC, nucleus
length 5.4 5,4 5.0 5.5
(.52) (,46) (.65) (.(,7)
width 2.3 2.3 2.3 2.5
(.25) (.39) (.28) P6)
10.5 10.6 9,4 11.6
(1.5) (2.1) (1.7) (2.6)
Macrogametocyte N=75 N=150 N'"159 N '"H(I
length 16.1 16.5 14.4 15.4
(.80) (3,4) (1.2) (1,4)
width 2.0 2.0 2.4 2.7
(.38) (.88) (.53) (.67)
41.7 39.4 42.0 44.5
(4.8) (7.9) (5.7) (5.1)
%Area of erythrocyte-
parasitecomplex 64.4 62.1 63.1 66.S
Macrogametocyte nucleus N<75 N:: 149 N'"159 85
length 2.0 2,4 2.6 2,4
(,4 1) (.60) (.54) (.52)
20 TAUI.F.2 (eonI'd). Morphometric parameters of thehaemoprotetdsof the Fringiliidae and Emberizidae.
H. frillgillae Hscoatneyi H. maual H.cnlons n.sp.
width 1.9 2.1 2.1 2.0
(AO) (A8) (.37) (.57)
3.3 4.4 4.6 4.1
(.69) (1.6) (.88) (1.2)
%Area ofgametocyte 8.0 11.3 11.1 9.3
Pigmentgranules-avg. 12.8 11.6 to.5 12.5
(3.0) (2.9) (2.2) (3.0)
range 8·21 7·20 6·17 6-21
Nuclear displacement
rutin .95 .78 .74 .66
(.15) (.20) (.18) (.20)
Erythrocyte infected
bymicrogametocyte N=27 N=15 N=3' N=lO
length 12.5 11.6 12.3 12.7
(.63) (.55) (.82) (.73)
width 6.1 5.9 6.5 6.1
(.63) (.51) (.46) (.44)
63.6 57.1 64.8 63.9
(9.3) (7.0) (5.3) (6.2)
21 TABLE 2(eont'd). Morphomet ric para meters
or
the bacmoprnrctdsor
the Fringill idaeandEmberi:tidne.H·frillgif/(lf! H.CO(l(IIl')'j lI./JIflZ!t/; II.chl{lri.~
n.sp.
Infectederythrocyte
nucleus N·27 N'"15 N'"34 N=IO
length 5.5 5.4 4.6 5.2
(.48) (.33) (.47) (57)
width 2.3 2.0 2.3 2.3
(.24) (.24) (.2H) (.24)
10.7 9.2 H.7 10.1
(U) (1.3) (1.6) (1.6)
Microgam et ocyte N ;;:27 N=15 N;;:34 N= lO
length 163 15.3 14.2 16.9
(U) (.7H) (1.2) (1.5)
width 2.0 2.0 2.9 2.6
(.39) (.83) (.73) (.KI)
41.0 37.9 43.9 465
(7.6) (9. 1) (4.3) (5.9)
% Area oferythrocyte -
parasite complex 643 65.7 67.9 72.7
Microgametocyte
nucleus N
=
27 N=
15 N=34 N=lOlength 6.4 6.7 6.6 6.8
(.69) (.HB) (.97 ) (1.5)
22 'fABLE2(cont'd). Morphometric parameters
or
thehaemoprote ldsor
the Fringill idaelindEmberlzldae.H.friflgillae H. coatneyi Hcmazzai H. chloris n.sp-
width 1.8 1.8 2.6 2.5
(.69) (.46) (.39) (.58)
11.1 12.0 15.3 18.8
(2.6) (3.1) (2.4) (6.8)
%Area of gametocyte 27.1 31.8 34.9 39.4
Pigmentgranules 13.8 9.7 11.7 11.2
(3.0) (1.8) (2.7) (1.8)
runge 9-21 7-14 7-17 9-14
Nucleardisplacement
ratio .94 .72 .68 .67
(.13) (.21) (.16) (.17)
Linearmeasurementsare givenin Jlrn,area measurementsinJ,tm1• 'Standard deviationsgive n(in parentheses)belowmeans.
:!J occupying 8%ofparasite. usually adjoininghostcellmargin.
Micrngnmer ocyte(PlateI.Fig.3; Table2): I laltcr idial parasite uf moderateIII larg e size. not usually causinghypertr ophy. cytoplasmfinelygranular.~taininglighter than mac ro ga metocytewithGiemsa's stai n; pigmen t granulesround tooval , some timesrod-shape d.yello w-brown. averaging14(rangingR-21);volutinrarely seen; parasitenucleuslarge.diffuse. stai ning verypate pinkwithGicmsn'sstain.
occupying 27%ofthegametocytearea.
Geognm hjc Ran ge:UnitedKingdom. Czecbostovukfu,Lithuania, Norway,Portugal, France.Spa in. andpresu ma blythro ughout thegeog raphic rangeoftheFringillldne.
Ba sisofredescription'
Neohapa nto type:Itis not knownifLabbe designated any hapantorype material.The neoh apanto type shouldbefromthesame host and local itya.s the originalbutifnu suitablematerialisavailable from eitherof these.a specimen fromthenear est locality andclosestrelated species maybedesigna ted asthenc ohapont ct ype(IC.l N 1985).Bloodsmea rsfrom thetypehos t anti locality were of poorqualityand were notsuitable forneohapantorypedesignation.Intheabsenc eofthe originalmaterial, thebloodfilm#92411fromFn'ltgill acoelehs,collectedbyPeirce atBramley,lIams, u.K.on June28,1981,isdesignat edas the neohapuntotypeofH./riflgillae.
24 Parnncobapa DIQlype: FromFringilla coelebs.Bloodfilm#67603 collectedbyJ.
Kucera on 21 May, 1975,Prague,Czechoslovakia.
Additio nalH05~:Appendix A.
C2..m..m..mL5: Labbe(1894)described a varietyofHalteridiumDanilcwskyi,whichhe slatedwas very common inthe blood of the chaffinchFrillgilla coelebs. The description.which included colourplatesbutnomeasureme nts, was ofasmall haherklial parasite(occupyinglessthan 50%of the infected cell area) which sometime shad atendency to surround thenucleus,with Indentatio nsalongthecell margins.Itwas laterraisedto specificstatusbyWoodcockin19 10, wholistedit as Halteridlumfringlllae(Labbe). Accordingto ArticleSt(e )(i) of theICZ N thisis only a change in rank withinthe speciesgroupsandtherefore did not require the use of parentheseswith the authority.
It isdifficult to determine whenHalteridium was synonymizedwith Haemoproteus,and when the designationHaemoproteusfringillae (Labbe) became correct. Haemoproteuswas first describedby Kruse in 1890,and as early as 1904 SchaudinnrecognizedHaemoproteus asthe validgenus,butfor yearsthe nameswere used interchangeably. Coatney (1936) statedthatHattaidium wasone of the synonymsofHaemoproteus,andspecificallylistedHaemoproteusJringiflae in Fringilla caelebs (sic).Sincethen, the nameH.fringillaehasbeen published as thehalteridial haemoproteidoccurringin many avian familiesotherthan the Fringillidae, from
25 whichit wasoriginally described (Ben nett. whtteway and wood wonb -Lyrue 19K2).
Inpractice,thetermH.[ringillae/ori:il'oroc'complex'(Peirce11)83)W(\Sused to identify thehaem opr ote ids ofmany host familiesbecau seofthe freque nt occurrenceof both smallhalteridlal parasites('fringillae-like').InUlarger,broade r par asites sim ilar10H.oru lvoracofthe famil yEstrlhl hfue. ItW;ISdifficultIn determine whether mixedinfections orone pleo morphicspecies were present .II.
orizivoraehasrecen tly been redesc rfbedand its runge redefine d(Bennett and Peirce 1991) .
Peirce (1984)red eseribedH.[rincillae,and distingui shed itfro m1-1.onzi\'(Jf{1(' on thebasis ofsmulle rsize,fewerpigmentgranulesand lessdisplace mentofthehost cell nucleus.However , themate rialusedintheredescription (hut notdesignated:IS aneohap ant otype) wasfromSylviaborinfrom Zambia.Thishost bird speci esisin thesubfa mily SylviinaeoftheMuscicapi dae.Thehaemoprotehlsofthisfamily huvc recentl ybeenreviewed and theHaemoproteusspeciesoccurring in the Sylviinac describe dasH.5J!/vae(Be nnettetal:1991).
In1916,deMell o and Braz de Sa descr ibedHaemoproteu s II/onumyfro m Copsycuussaulans(amember of thethrushsubfamilyTur dlnae of theMusclcapiduc).
Bennett andCampbe ll (1972) synonymized(ill po/tim)H. momonywhhl-l. f rillgilfflf!
based ontheearlie r descripti on and figures. Howeverthehaen.oprotci ds of the thrushes haverecently beenreviewedandredescribcd, withthe haem ap rntc id species occurring in thesubfa milyTurdin aerefer red 10H.[GII/isi(Benne ttcl al.11)1)1).
Inview ofthe host family andsubfa mily specificity experimenta lly
26 demonstratedforanumber of species ofHaemoproteus,and the resultsofthisstudy.
assigningH.fringitlaetosuch a broadrange of hostsis probably inerror. This species sho uld belimited 10the Fringillidaeuntilexperimentalevidence dearly indicatesa wider host spectru m.In this study,H.frillgilluewasidentifiedinFringilla coelehs and F.montifringilla.No materialwasavailable fromFsteydea,the onlyother representativeof this family. Haemoproteus fringillae ishereinredescribedwith neo hapantotype and peraneohapantotypeslidesdesignated.
Haem oproteusfrillgillaeis amediumsized halteridia!haemoproteidwhich typically neitherdeformsthe host erythrocyte inany way, nor displacesitsnucleus, and whichoccupies approximately65%of the erythrocyte -parasit ecomplex. The nuclei of both macrogametocyteand microgame tocytearetypically smaller than that of otherhaemoproteldsin relatedhostfamilies(Table2). Theva ria bility in pigment granulesizeand numberiscomparablewith that seen in otherspecies.
17 EMBER IZ IDAE
Emberjzl nne
Nomennudu m: HaemoproteusclIllleriw
H. emberizahas been variously listed inthe Emberlzinue,byZeinicv(197~).
Musaev and Zeiniev(1977),andSubkhannvand Mlrzobakhaduro v(1976)who attributedthe species to Berson(1964). Berson,howeverdocs nor cite thispnnettc species inhispaper,and there is no descriptionof the species. PeirceandBenn cu (1979),in describingexamplesof problem sarisingin the pastin Russian tltcnuurc.
designatedH.emb eruaasaIlO lII e lllnudum,
Haem op roteus cOll1neyin. sp,
Type Host:Zonotrichiaalbtcottls(Gmelin) (White-throated sparrow) TypeLocalit y:Gande r,Newfoundland,Canada
Immaturegametocyte (Plate2. Figures 1-2):Youngest forms seen usuallyinitiate growthlateraltoerythrocyte nucleus,although polarpositionsaresometimesfound;
parasitessmalldu mbbell-like or sausage shaped andusuallyapprcssthehos tcell nucleus; marginsusuallyentirewith ameboidedges common only inintense infections.
MAcrogam etocyte (Plate2, Figure 4-7;Table2):Halteridiul purusiteofmedium In
PlATE2
Haemoproteuscoameyin,sp.
Figure)-2. immaturegnmetocyres Figur es 3. rnlcrogume tocytes Figure 4-7. rnacrogumetocytes Figure R. rntcrogaruetocyte
1
C'-Y
2
8
29 large size, dumbbell-shaped.or broadly haltcridial, occupying 55-70% ofthe host erytbrocyte.parasite complex;hypertrophy of infected cell area variable, (rom 2- 25%.sometimesdisplacingthe erythrocytenucleus. usuallyonlyinsausage -shaped
forms(NDRra nges from0.5to1.0)(Plate 2,Figures 5,7); cytoplasm granular, staining moderate blue with Glemsa's stain; marginsentire and usually rounded;
gametocyteusually appressed to the erythrocyte nucleus,but frequently the outer edge doesnot appear to extendto the outerperip heryof the erythrocyte.causinga 'dip' andgiving a hilobedordumbbell appearance (Plate2, Figure4); pigment granules. whenvisible.appear eithe r lar ge and rod-shaped, smalland ovoid,or irregular,yellow to darkbrown, averaging 9-13(ranging9-21),usually distributed randomlythroughout the cytoplasm.but in some hosts (especiallyPipi/ospp. and Sporophila spp.)are paired; volutingranu les no tusually seen; parasite nucleus round to broadlytriangular, termina lto subterm inalin position, occupying1().16%
of the gametocy tearea; multipleinfections of 2-4parasitesin intense infections(2 most commonof multiple infections).
MlcrogumctQcyte (Plate 2, Figures 3,8;Table 2): Halteridialparasite of medium to largesize, occupying66% of thehost erythrocyte-parasite complex; hypertrophy of erythrocytevariable.rarely displacingthe erythrocyte nucle us;cytoplasmfinely granular,stainingonly lightly withGiemsa's stain; marginsentireandusually rounded; pigmentgranulessmallto medium and ovoid or irregular,oftenclumped ut poles of parasite:volutin granules not usuallyseen;parasitenucleus large and
30 diffuse.oftenno t distinguishablefromthe cytoplasm. stniningpalepink with Giemsa's stain,occupying32%oftheparasitearea.
Geogra ph ic ranee: North nodSouthAmer ica.Patecr ctic.Africa,100..Ii:l.A"ia.
presumablythrou ghouttherangeof theEmberizinne.
~: Bloodfi:mII79654fromZonomchiafll/Jicof/i J,collectedhyJ.nurry Cainesat Gande r,Newfou ndlandonAugust18,1979.
Parnh apnntotypes: Blood film/I30671from Pa...sereua iliaca,collectedhya.F.
Bennett atPickavance Creek,Newfoundland. CanadaonJune 14.1972; From Zouotrichiacapensis,(I46137 collectedbyR.W.McFarlane at Arlee,ChileonApril 28,1973,1/7665 4collectedbyO.SouzaLopes athapeu nlnga,Brazilon April3.
1969.1/ 97246 collectedbyA.Gabaldonat Merida,Venezuela on Septem ber4.
1980; # 97846 fromZonoeichiaalbicollis,collectedbyC. Kirkpatricc atMercer County,NewJe rseyonMay 5,1985;# 113646fromPasserellailiucu,collectedhy P.E.SuperatPo intReyesPeninsula,Californian,'Septe mber3D,191'19.
Additionalrecords:Appe ndixA.
~:Hae moprotel ds were first discovered in theblondof No rthAmerican spar rowsofthe genusMelospua ,byOplein 1898. Amongothers,Novy and
31 MacNeal (1905)observ ed theminMelospua andSpize11a,and Manwell andHerma n (1935)listed theminPasserculus,Junco, Spizella,and Melospiza.Howeve r,due in part10poordescriptions andthe variability observed,most authors were reluctan t toide ntifythe speciesofHaemoproteus,and identifie d them simplyasH.sp.
(Manwell1955,Bennettor01.1982). Variousstudies have referred the names Haemoproteusfrillgillae.H.otiuvorae or H.fdngillaelodzivoraecomplexto those haemop roteids havingthe samebasi chalteridlal form (Bennettet al. 1982,Peir ce 1983.1984).Thesedesignationshavebeenmadefor hae moproteidsof manyhos ts includingthe Emberizinaein the absenceof com prehensive studiesof theparasites acrosswidehost andgeographicranges. Some parasitesweremed ium-sized halterldial forms,often appear ing bilobed.Othe rs wer elarger, broaderandoften displaced theerythrocytic nucleus. The numbe rand sizeof pigmen tgranuleswere variable.Whetherthere wereindeed two sepa ra tespecieswas confoundedby the frequentoccurrenceof buthtypesin anyone blo od smear.
Material from 25host species across a widegeograp hic rangeexamin ed inthe curre ntstudy (AppendixA)revealedthatther e was variabilityinthe shapeof the haemoproteldsinthis subfamily,andthat pigmentin mature gametocytesvaried from a few(7-10) lar ge gran ules to several(17-20)smallormediumgra nules. In many smears,thepigmentwas notvery distinguishableand ther e wereboth parasites with small pigmentgranulesas wellas those withlargeelongate rod-shaped granules,and indee dmany withboth sizes.Therewasno consistentpatte rnof pigmentgranule sizeandnumbe rwiththe overall morphology. Also, inmany cases therewere
32 specimens of both thedumbbell type and thebroaderhalterldiultype regardless of whethersmearartifactswerebelievedtobepresent.In any onesmear,therewas notalways acleardistinction of infectionwitha dumbbellshape (witha'dill')(Plate 2, Figure 4) andthatwithalarge broader form(Plate2,Figures 5-7)butrnthcr u continuumexisted (PlateA.Figure 3).
Smears from individualbirds takenat differenttimes showedrmpaucrnofthe occurrence of onetype ortheother.For example,oneZO/lotriclliaa//Jicolli.ffrom Newfoundlandshowed aninfection of almostall dumbbell forms inlute May (pre- transmission season, therefore probablya rela pseinfec tion). OneweekInter hoth dumbbell (so mewith 'dips')and broade rformswereseen, and two weeksrater (mid- June) only broad forms werepresent. Another Z.albicollis.from NewBrunswick, showedthereversepattern in three captures beginningin early June. Whether or not progressivedevelopmentof gametocytesresults insuchobservations can only he det erminedby life cyclestudieswhere birds are not subjec tto reinfectionas they arc inthe field. In any ca se,throughoutthewholehost andgeographic rangeexamined, there wasfreq uentthoughno t totallyconsiste ntoccurrenceofthe 'dip'. Itwas especially pronouncedinSouthAmericanspecimens(especiallyinZonouichia capensu)where theparasites were fairlysmallcomparedto somc from North America. Sometimes it appearedthat thecytoplasm simply didnotstain intbc centralport ionof the parasite,butinother cases,a distinctcellmembranecouldhe seen.Often,'fringillae type'parasitescanbe see n ononeparticular sectionofthe smear,andthe'cdzivoraetype'on another.The fact that thisdiunotoccurin just
33 a few isolatedcases points to either two species transmittedby the same vector, or nne morphologicallyvariablespecies.
Culicoidescrepuscularisand C. sphagnumensis havebeen identified as containingsporozoltcsofthehaemoprotcid foundin NorthAmericansparrowsof the genusZonooicnia(FallisandBen nett 1961). Khanand Fallis(1969) described the schizogonicstagesin lungtissue ofZOllo /rielliaalbicollis.However,the specieswas ide ntifiedasH.fringillae. In view of the currentstudy's findings and inaccordance withthe currentlyacceptedfamily/subfamilyspecificity(Benneuand Peirce1988) thismiside ntifed specieswas almostcertainlyHaemoproteus coatneyi.
Haemoproteus coatneyin. sp. is hereinproposedand describedas anew speciesfoundin the subfam ilyEmberizinae of theEmbe rizidae,andbaparuotypeand paraha pantotypeslides designat ed. The specificnameisin honour of the late G.RobertCoatney,inreco gnition of his contributio n tothe study ofhaemat ozoa of NorthAmericanpasserlnes.
EM RERI?..IDAE
~
Haemoproteusmazzai Parodi andNino,1927
Synonym: Haemo p roteus IledymelisCoatney and Roudabush.1937
Type Host: Pheucticus aureaventrisd'Or bigny andLafresnayc [Black-backed grosbeak )
TypeLocality:Louisiana,U.S.A.
Imm3l U~(Plate3, Figures1-2):Younges t for ms seen lateralor medial
to the host erythrocy tenucleus,or rarely,at eitherpole;gametocyteusuallygrows withmembrane appressed to the host cellnucleus,developingfromsmallovoid formstothin bana na-shapesto thickenedsausage-shap edforms;marginumocr okt orentire.
Macrogametocyte(Plate 3,Figures 3-5;Table 2): Halteridialpa rasite ofmedium tolarge size, sausage-shapedorbroadlyhalteridial, occupying 60-70% ofthehost erythrocyte-parasit e complex; hypertr ophyinareavariable,(5-25%);displacingthe nucleusinsome cases (usua lly about half);cytoplas m fine ly granula r,occasionally vacuolate ,staining modera te blue withGiemsa's stain; marginsentireor with indentationsatends, appea ring amoeboidor square,especia llyinthosenot quite
Haemoproteusmarzai
Figure1·2, immaturegametocytes Figures3-5. macroga metocytes Figure6. microgame tocyte
5 6
36 fully matur e: ends of parasitesofte nnotreaching poles ofhost erythrocyte,usually touchingcell nucleusbut notalwayscell periphery; pigmentgranules large androd- shaped, orsmalland ovoid. paras ites with bothsizes see nin the samesmear,yellow 10 darkyellow-brown,averaging11-14 (ranging7-16)distributedrandomlythroughout theeytoptesm,sometimespairedorinsmall clumps;volutin granulesseen onlyin parasitesofaCardinalispnoeniceusfrom Venezuela;parasitenucleusround ormore
rarelybroadl y triangula r,medi alto sub-me dial in poshia n, butoccas ionallyter minal, located on theouter marginoftheparasite,rarelyappressedtothenucleus;
occupying12%oftheparasitearea;multip leinfections(1-2game tocytes)of the sameerythrocyte see n in intens einfectio ns,usuallyonoppositeside sof the host cell nucleu s.
Mlrrogomf locyte(Plate3, Figur e 6;Table2):Halteridial parasiteof medium to large size,occupying60-70%ofthe host erythrocyte . parasitecomplex; cytoplasm finelygranular,staining onlylightlywithGiemsa's; endsusuallyrounde d and margin enti re but sometimes amoeboid; pigment granulesusuallysmallandovoid,evenin thosehosts wheremacrogemetocytes havemostlylargepigment granules,polarin distrihuricn,aver aging 10-13 perparasite:para sitenucleus largeanddiffuse,sta ining verypalepinkwithGiemsa' s stain, occupy ingapp roximately 35%of thepar asite nrcu: volutin gra nulesseeninpar asitesofaCerdinalisphoenlceusfro m Ven ezuela.
Grogrnobicmngr: NewBrunswick,Canad a;California. Maryland,NewJersey,
~7 U.SA;EI Salvador ;Jamaica,Venezuel a andpresumablythroughout theentir e range of thesubfamilyCardinali nae.
Ba si, ofre4escrip lio n'
NeohoDanlQrype:Inthe absenceofsuitable materialfrom the origina l typehO,"1arul lo cality,Blood film/I103795 fromCordindiscardinolis,collectedbyM.Garvin ut BatonRouge, Louisiana. U.S.A. on May 16, 1988 is designated :t~ the neohapantotypeofHaemoproteus",(Uza!.
Paraneohapanlolyocs;FromPheucticus ludcvicianns:Bloodfilm/I57493.II 60293.
II60384.#60385. collect edbya.F.BennettatTantrumarMarsh.New Brunswick, Ca na daonAugust5.1976,June 29,1m.July 5.19nand July 5,vm.respectively;
/I98013. collectedbyCKirkpatrick atNewJe rsey, U.s.A. onMay1J,19M;If 113671,fro mPteu akusmekmocephalus,collectedbyP.E. SuperatPoint Reyes.
Ca1i£ornia,U.s.A . onMay3,1990.
AddjHQnjJlmQrds·Append ixA.
~: Parodi and Nil\o (1926) described, and lnter (1927) named Haemoproteusmazzaifrom both stainedandfreshblood preparationsandlungtissue sta gesfoundinP/Jcllc,icusQurCO\1elltrisfromnorthernArgentina.No hapanrotypc materia lwasdesignated .The authors describedtheimmat uregamctocyucfllrms
38 as "more or less globular or in the shapeof small bananas'andthematur e parasites as "widened halteridialformswithnumerous granulesof pigment..."which sometimes caused nuclear displacement,and eventuallyruptured the host cell. They did not give specific pigment granu lecounts butstated they were "irregularof form,attimes distributedaround the nucleus and at othertimes at the extremitiesof the parasite".
Their plates andfigures indicated a broadlyhalteridial or sausage-shaped parasite similarto those seen in the current study.The figures showed fairly large pigment granules.numbering from 8-14, The description included thatof schizonts in endothelial cells of lung alveoli. However, thestatement of finding mcroaoltcsin leuco cytes as well as the endothelialcells, and "numerous melanic monocytes' is confusing. These findingsmustbein error as merozoites of Haemoproteus are not foundin blood cellsother than erythrocytes.They describedthe developmentofthe parasite as being similar to that ofH.orizivorueandH.cotumbae.
Coatney andRoudabush (1937) described, fromPlreucticus ludovicianus,a halteridial haemoprcteld which they called Haemoproteus Izedymefis, and distinguished it fromH.mazzaiby the former's extreme nuclear displacement and tendency to become round and burst out of the host cell. This rounding and 'bursting out'of the gametocyte was likely either the typicalpost mortemchanges that occur in haemoprcteids or the result of slow-dryingblood,and are considered of no taxonomic consequence (White and Bennett 1979, Bishop 1989). Coatney and Roudabush describedH.IJedymefisas generally having smoothrounded ends,usually touching the hOM cell nucleus, but not always the cell periphery. The
:w
macrogametocytes usua lly caused some nuclear displacement while thc microgemetocytesdid only in about half the cases. The pigmen t gra nules were ellip ticalto rod-shaped,ranging from8-12(average9).Fromthe description and figures giveninboth descriptions, it is clearthattheyarc thesame species.
Examinationof materialfromPneuaicusandCordinaliscon firmedthe occurrence of one speciesin this subfamil y,with a pleomorphicform. Bythe Princip leor Prior ity(Article23(a) of the ICZN).H.mnaaiParodiandNino1927is thevalid species name. H.hedymelisCoatne yand Roudabush 1937 isher ebydeclareda syno nym ofH.maaai,
The materialexamined in thisstudy fromthis subfamilywas fro m threehost species from a varietyof locations; the suhf amilyislimitedlONorth andSmull America. Haemoproteusmauaiexhibitedabreedrangeof size and shapeundis fairlytypical ofthemoderatetolargehalteridial baemoprotctd.Theinconsistency ofga me tocyte outline, nucleardisplacement and pigmentgranulesizewer epresum ed to be the result of natural variation.as any singlebloodsmearshowed a v••rtctyof forms . Althoughother South Americanspeciesofthisaviansubfamily have been examined for blood parasites(woodwcnh-Lynasat al.1988), there ure no other records ofHaemoproteus(Bennetter at.1982).The host speciesandgeographical locations from whichH.mazzaiwas found in the current study aregivenin Appendix A.Haemoproteusmastaiishereinredescrlbed asthehae moprotel d speciesfound in the Cardinalinae of the Embe riziduc, and ncohapa ntot ype and paraneohapantotype slidesaredesignated.
40 EMRF;RIZ IDAE
~
Nomina nuda: Haem oproteus loxiaeTart akovskil, 1913 Haemoproteus acandusMusaevandZeiniev, 1977
In 1913 atalaboratoryexhibitioninPe trograd(Leningrad), Tartakovskii describedand illustratedwilh tablesand wetercolour pictures,five speciesof Haemoproteus.AmongthesewasHaemoproteusloxiaefro mLoxiacurvirostra. The procee dingsfromthe exhibitionwere published.hutthey did notcontainanyofthe illustrati o nsornumeric aldata.Th e mate ria l,storedinthelaboratory'sarchives, was used asfuelin the siegeof Leningrad duri ngWorldWa rIf(PeirceandBenne tt 1979). Therefore,it isnotpossibleto comparethepictures with any subseque nt material from member s of thissub family.Tarta ko vskii'stext doesnot meet the basic requirem e nts of theInt ernationalCode ofZoologicalNomenclature(tCZN1985) forspe cies des ignation ,and Peirce andBenn ett (1979)de claredthe speciestobea nOli/eftnudum.
Musacv andZeiniev (1977) listedHaemoproteusacamhis as a parasi teof Acanthls cannabina. However,they did notgiveany descriptio norillustrations,and infact spelledthespecific namediffere nllywhen theyliste ditin text(H.acan thies ).
Again, this docs notmeettherequirement s of theICZN as a speciesdescrip tio n, and thereforeH. acaml,iswas declaredanomennudum byPeirce andBennettin1979.
41 HaemoproteuschIom Coveteon Ort ega andG611cgo ucrcngu er,1950
Synonyms: Haemoproteusg/obulosusC. Ortega and G. Berenguer,[950 Haemoproteus macmplgnientatus C. Ortega and G. Berenguer,19S0 Haemoproteussaini Levineand Campbell,1971
Haemoproteus wrrakovsJ..)'i vatkfnnas,1986
Type Host :Carduelischlons (Linnaeus) {European greenfindlJ Type Locality: Malzeville,France
Immature gametQcyte (Plate 4, FigureI): Youngestforms seen latcrul OTmedialto the hosterythrocyte nucleus, rarely at pole; marginamoeboid or entire.
Macrogametocyte (Plate 4, Figures 2·5;Table 2): Haltetidialparasitera nginglmm alarge broadlysausage-shapeor amedi um-sized dumbbellshape, occupying 60-75%
of the host erythrocyte-paras itecomplex, at timescnusingS- IO% hypert rop hy in area.
especiallyin NorthAmericanhosts; displacing the nucleus in some cases and often causing atrophy of the erythrocyte nucleus: cytoplasm granular, often vacuolate, staining moderate blue with Giemsa'sstain; margins entire and rounded although sometimes withindentatio ns at ends, appearing'squurish':pigment granules usually large,often rod-shaped,or small andovoid, often both sizes in one gurnetocyte, yellowto darkyellow-brown, averaging 14 (ranging 9-25 with host and geographic
PLATE4
Haemoproteus cli toris
Figure1. immaturegametocyte Figures2-5. mncrogametocytes Figure 6. rmcrogametocyte
2
4
5 6
43 variation), distributedrandomlythroughout thecytoplasm,and morerare lycl umpeJ;
volut ingranulesnot usuallyseen; parasite nucleus roundtoovoid or broadly triangular, medialto sub-medial in position and usually locatedonthe outermargin of the parasite,occupying 9% (ranging5-12%)ofthe parasitearea;multiple infections(1.4gametocytes)of the sameerythrocyteseen in intense infectio ns, sometimes on the samesideof thebe stcell nucleus,(seenmost ofteninLoxiaand Acaruhisspp.) causingextremenuclearrotationand/ordisplaceme nt.
Microgomelocy1r (Pla te4,Figure 6;Table 2):Halte ridial parasite ofmedium 10 large size.occupying73% ofthehost erythrocyte. parasite complex; cytoplasm finely granula r,staini ngonlylightlywithGiem sa's stain ;ends usuallyrounded and margin entire nut somet imesamoebo idorsqua rish; pigment granu lesusuallylar ge androd-shapedbutalso smallandovo id,usually clumpe dnea rthe poles,averaging 1t-14per parasite; volutingranules not usua llyseen; parasitenucleu slarge and diffuse, stain ingpalepinkwithGtemsa'sstain,occupying approximately30% of the parasitearea.
Busisof re.d(!:grl ptlon :
NcohaPontQhp(!::Itis not knownifCovaleda Ortega and GallegoBeren guer (1950) designatedhapantotype material,or whethertheiroriginalcollection isintact.
According totheICZN , the neohapantotypeshould befromthe samehost and localityas theoriginalbutifno suitablematerialis availablefro meithe rof these,
44 aspecime nfromthe near estlocality andthe closestrelate dspecies lIluy he designated as the neo hap antotype. Nosuitable mate rialfromthe typehostand local ity was availableforstudyordesignation. Intheabsence of 111('nrigiunl materia l,Bloodfilm# 68217,fromCarduelis cardudis, collecte dbyJ. Btnncouat Mulzeville,France on September6, 1978 is he re by designat ed as the ncohupunuuypc ofHaemoproteus eli/oris.
Pnrn nCQhnnonlotypcs: Bloodfilm #21924 fromLoxiacurvimstra. collectedby a.F.
Bennett atPlckavunceCreek,Newfcunulnnd,CanadaonJuneHI,1971;#51'1131 fro mCarduclis 'MOri.f,collectedbyM.A.Peirce atEngla nd onMay21,1% 5; # 107373fro mAcanthisflulIllllea,collectedbyG.Scurin at Churchill.Manitoba, CanadaonJune24, 1989.
Addilion al ho st records :Appen dixA.
GeognlDhjcRange:North America,Europe,Asia, Africaand presu mably throughout theentire rangeof thesubfamilyCardu elinaeoftheEmberi zldu c.
~: Covale daOrtega andGallego Berenguer(1950)described three species of haemoproteids frommembe rs of thegenusCarduelis.Haemoproteusg!ofJII!o.uu was describedasaha lter idia l parasitethatwhen mat ur e lookona globular form.
displacingthehostcellnucleu s toone marginandcausinggrea thypert ro phy.The
45 descripti onwa.sbarsed solelyon onepositive bloodsmear andfromthe description and figureswasslikely theresultofusing bloodfrom a deadbird or from slow drying of the smear,twoconditions whichare knownto causesuch artifacts (Bennettetal.
1975,White and Bennett1979). These forms represent the initiationof gamete formationthat would no rmally occuruponingestio n by a susceptiblevector. The fact thai'imma ture' halleridial formswere seenin thesmear further supportsthis view.
Peirce (1983) alsobelievedthistobethe case but didnotsynonymizethe name of the species.Alsoin 1950, CovaledaOrtegaandGallegoBerenguerdescribeda similarglobularspeciesoccurring inowls. This species was also presumedto be describedfrompostmortemmaterialcausing changesin the gametocyte and has since been synonymized(Bisho pand Bennett 1989).Conside ringthis.andbecausethe presumedinfectio nwas found inonlyone bird,H. globt/lasusis hereby declared a :.)'nonymofH.chloris.
CovaledaOrtega andGallego Berenguer(1950) alsodescribedHaemoproteus mac:ropiglllentu,"sfrom one smearfromCardueliscorduetis. The parasitewas described as beinghalteridialwithvoluminousappearance whichcauses slight displacementandrotationof the erythrocyte nucleus. They describedthe pigment aslarge oval-rounded dark viole tgranules,sometimessurpassing1umand numberi ng 7-11 in macrogametocytes,andbasedthe ir diagnosislar gely onthese characteristics.Itappearsthatthey confused volutingranules(which are purple-red in color),u fairlyvariablecharacter isticof someHaemoproteusspecies (oftenwithin :10individual hosl bird) withtruehern uzoin pigmentgranuleswhich areyellow-brown
4"
(Bishop 1989. Bennettand Pei rce1989). Vol utinwas only rarelyobse rvedinthe carduelinematerial inthis study.Allothercharucterisiticsof this described specimen were thesame as thoseofthebasic halteridialparasitesseenin theothersofthis subfamily.H.macroplgmenunusishereby declareda synonym of fl.cnlons.
Valkiiinas (1986)describedH.tartakovskyifrom Loxia curvirostrninLithuania asthefirst 'enucleat or' typehaemo proteiddescribedfrom thepnsscrlncs. The immatureparasitesaTCcomparab leinbot hsizeand shapetothe haltcrkli:\1form seen inothermembe rsof theCarduelinac.The major differ e nce was the occurre nce of.somegametocyteswhich displacethenucleus, <IOUsometimescnuxingrotation of the nucleus.A~well,some occasionallyenucleatedthe infectederythrocyte.l11CSC characte risticswereseen in the currentstudyin materialfromLoxiaandAcanthisin North America,althoughenucleat ingforms were notasfrequent asinVlllkiiinas' mater ial.Inaddition,periphe ral blood smearsfromanybirdspeciesoften contuin afewspecimens that showsimilarnuclear displacementand/orrotatio n,usually aI the periphery ofthe smear.Th ese are probablycausedby excess pressureon the blood asit is smeared on themicroscopeslide.Other recognizedcnuclcutnrtypes (H. enucleasor,H. bel/ncui)show afrequent occurrenceof the displacedand enucleatedspecimens. InH. benneni,thepercentage of parasitizedenucleate dcells variesupto 96%(Grein er eral.1977). Inspecimensofcarduclinesfmm North America,theoccurrenceofenucleatedcells is minimal«2%).SeveralsuucsofII.
tarkakovskyi in Loxia curvirostru,from Lithuania(includinga paruhupan totypcslide]
were examined inthisstudy. The parasite was gene rallylarge andoftenshowed
47 extreme nuclear displace men t,andthe rewereinstancesof enucleation.Howeve r.
in many cases,multipleinfection,seenoften in intenseinfections,caused the displacement. Two ormoreimmature gametocytesinitiati nggrowthon the same sideofthe nucleusresulted in displacementandrotation (PlateA, Figure4).This couldbemistakenfor truedisplacementcausedbyone gametocytein some cases where differentiationoftheparasitemembrane was difficult. Atthistime, until experimen talevidencecan confirmIhe existence of a differentspecies,the haemoproteid occurringinLithuania wouldbe better considereda geographicvariant of asingle.pleo morphic species.H.tarkakovskyi isher eby decla reda synonymofH.
chlons.
Sergent and Sergent (1948)describedaHaemop roteusspecies from a canary tSerinusCfll/aria)thatwas held in captivityin Algiers,Algeria, in the sameareaas Algeriansparrows (family Passerldae)fromwhichH.wcnyaniSergentandSergent 1948 was described.ThisHaemoproteuswas similarinappearancetoH. wenyoniand allho ughSergentandSergent described it separately,theydidnot give it ana me.
Levine andCampbell(1971)assignedthe nameH. serini,based upon the parasite's occurrenceina separategenus.111eydidnotred cscrihe thespecies, however.The na meH..serinithus datesfrom1971, whilethe descriptionisfrom1948. The haemoproteid examinedin thisstudyfromcanarieswas consideredthesameasH.
chloris, which holds precedenceaccordingtothePrinciplesofPrior ity ofthelCZ N.
Therefore.H.sainiis synonymizedwithfl.chioris.
Haemoproteuschloris is a halteridialhae moproteid witha broadgeographic
4X and hostrange.In the materia lstudied.it exhibited ahighly pleomorphic form, rangingfrom a broadlyfat sausageshapetoamedium-sizedhalteridinl parasite.
While the smalle rbalte ridialshapedidnot decidedlydisplace thehostcellnucleus, but rather envelopedit.the largersausageshapeoftendisplaced thenucleus towards the periphe ryofthe cell.Often forms ofthe entirerange were seenin anyone bloodsmear from anyonebir d.
In most ofthematerial examined.therewere usuallyparas iteswitheitherfew large rod-shaped pigmentgranu lesorsmallovoidones. However,there were specimensin all birdgeneraandgeographicareas with hothsizesof pigment granuleswithin the same gametocyte(PlateA,Figure 5).There wasnoconsistency in therelationship of parasitesizeand shapewith other generallyuxcdebnrnctcrs such as pigment number, marginoutlinesordegreeof nuclear displacement. 111e basictre ndof larger ratioofpamsite toparasite- erythrocyte complexarea...ithmore nuclear displacement wasthe same for allwithinthe subfamily,asmighthe expected in samplingparasitesatvariousstages ofgrowth anddevelopme nt.
The frequen toccurrenceof specimens in a singlesmearof bot hdumhhell shapedgumetocy-esandfullerhulter idiul gametocytes whichure not uttributublc to smear artifacts lends support for onehighly morphologica lly variable specieswithin this subfamily.Consideringthe large numherof host birdspecies(125)withinIhis subfamily and thelarge geogra ph ical range (fourcontinents)inwhichtheyoccur,this notsurprising,and mayillustra te a still-evolvingspecies. Life cycle cross-uunsmissto n studies may reveal whe the rsuchgeographicaland/orhost induceddifferencesoccur
49 ut thc specieslevel.
Haemoproteuschlorisishereinredescri bed as thesingle haem opr oteid spe cies found in the Ca rdue linae of the Emberizidae, and neohapantotype and paraneohapantotypcslides aredesignated. The specificnameis emendedfrom chkviis to chkxis inaccordancewithArticle34 (b)oftheICZ N (1985).Thespecies ispleom or phic and may representa complex of a still-evo lvingspecies.
Haemopro teus parufin.sp.
TypeHosu DendroicacorOllata (Linnaeus)[Yellow-rumped warbler]
TypeLocality: Gander.Newfoundland.Canada
ImmD!ure gametocyte (Plate5. Figure1): Youngestforms seeninitiate growth lat eral10 host erythrocytenucleus.usually ncar middleof nucleus: Margins entire.
Macr ogam etocyte (Plate 5, Figures2·3;Tnble3):Hultcridial purusitc of medium tolarge size,occupying-70% ofthe host-erythrocytecomplex;hypertrop hyof cell areavariable ,rangingfrom 2·23%.displacingthehostcell nucleusonoccasion;
cytoplasmfinely granular,staining moderatebluewith Giemsa'sstain;rnurglnsentire andusuallyroundedalthough cansometimes be squared;pigment granulesmedium sized,usuallyovoid, although can be rod-like, yellowto dark-brown,averaging 14 (rangingfrom7·25),distr ibutedrandomlythroughout thecytoplasm:volutin notseen:
parasitenucleus roundto broadly triangular,subterminaltotermi nal in position.
occupying 10% of thegametocytearea;multipleinfections(1.4) pa rasites seenin intense infections.
Mjcrogamel ocyte {Plute 5,Figures4·5,Table3):Haltcridia l parasite of mediumto
PLATE 5
Haemoproteus parulin,SlJ.
Figure1. immature gametocyte Figures2-3. rnacrogametocytes Figure4-5. mkrogumetocytes
