2003.v smith.dresden

Lousy Phylogenies:Phthiraptera systematics and the

antiquity of lice

Division of Environmental and Evolutionary BiologyInstitute of Biomedical and Life Sciences

Graham Kerr BuildingUniversity of Glasgow

Scotland, UK

Vincent S. Smith

Lice (Phthiraptera): biologyi. Diversity

• 4,384 spp “chewing lice” (3,508 bird & 402 mammal spp)• 543 spp sucking lice (812 spp mammal)

iv. Cophylogeny, cospeciation & coevolution

ii. Physical characteristics• wingless, dorsoventrally flattened, 0.3-11mm, 3 nymphal instars• feed on feathers, fur & skin debris (“chewing lice”) or blood• adaptations against host grooming

iii. Distribution• mirror host distribution worldwide in virtually every habitat• absent on Chiroptera, Cetacea, Microbiotheria, Monotremata,

Notoryctemorphia, Pholidota & Sirenia.

- 1,350 spp in 6 families- Most “primitive”

suborder- More vagile, less host specific- Widespread on birds- 2 fam. South American mammals & marsupials- 1 fam. on Australasian marsupials

- 3,060 spp., # families ?- Widespread, ~90% spp. on birds- Avian groups poorly known- Highly host specific- Niche restricted- Mammal infesting species of veterinary significance

- Just 3 species- On unrelated hosts- Acquired via shared use of watering holes?- Tiny mandibles present at end of a long rostrum pierce hosts thick skin

- 550 spp. in 16 families- Absent from birds- Well studied- Greatest medical & veterinary significance- Vectors of several important diseases- Low prevalence on healthy hosts

AmblyceraAmblycera IschnoceraIschnocera RhynchophthirinaRhynchophthirina AnopluraAnoplura

Lice (Phthiraptera): taxonomyChewing Lice“Mallophaga”

Sucking Lice(Anoplura)

Lice (Phthiraptera): phylogeny

PsocopteraPsocoptera IschnoceraIschnocera RhynchophthirinaRhynchophthirina AnopluraAnopluraAmblyceraAmblycera

Chewing Lice“Mallophaga”

Sucking Lice(Anoplura)

Kim & Ludwig, 1982(Morphology)

Parasitic Lice((Phthiraptera)

Lyal, 1985(Morphology)

GenusFamilySuborder

3

532

3,060

1,344

Rhynchophthirina

Amblycera

Ischnocera

Anoplura

MammalsBirds

Amblycera

Ischnocera

Rhyn…

Anoplura Phthiraptera:

Phylogeneticresolution, 1996

GenusFamilySuborder

3

532

3,060

1,350

Amblycera

Ischnocera

Anoplura Phthiraptera:

Phylogeneticresolution, 2003

Rhyn…

MammalsBirds

GenusFamilySuborder

3

532

3,060

1,344

Amblycera

Ischnocera

Rhyn…

Anoplura

1996GenusFamilySuborder

3

541

3,060

1,350

2003

Amblyceran Families

85

84

Grilicolinae

Gyropinae

Protogyropinae

Trimenoponidae

Ricinidae

Laemobothriidae

Boopiidae

Menoponidae

Liposcelidae (Psocoptera)

Birds Worldwide & Australasian Marsupials

South/CentralAmerican

Rodents & Marsupials

GyropidaeAnalysed by

Marshall, 2003from Clay, 1970(Morphology)

Laemobothriidae- Falconiforms

Menacanthus/Menopon-cpx.- Birds worldwide

Colpocephalum-complex- Birds worldwide

Austromenopon-complex- Aquatic birds

Dennyus-complex- Birds worldwide

Boopidae- Australasian marsupials

Ricinidae- Hummingbirds

Avian Amblycera

6457

8773

1

1

611

7561

3100 999

93853

10010012

10010015

9688

100 98

7

8

8976

4

3

5

9074

2

1

1 622

9896

11

1

11

1

523

2 80574

92755 51

13

1

2

1

2

1

2

2

1

12

Trochiloecetes rupununiRicinus fringillaeTrochiliphagus abdominalisLaemobothrion maximumTherodoxus oweniBoopia tarsataParaheterodoxus insignisLatumcephalum lesouefi/macropusParaboopia flavaRediella mirabilisActornithophilus uniseriatusPlegadiphilus threskiornisChapinia robustaBonomiella columbaePseudomenopon pilosumAncistrona vagelliDennyus hirundinisMyrsidea victrixHolomenopon brevithoracicumEidmanniella pellucidaMachaerilaemus laticorpus/latifronsAustromenopon crocatumNeomenopon pteroclurusHohorstiella lata

Osborniella crotophagaeEomenopon denticulatumPiagetiella bursaepelecaniCiconiphilus quadripustulatusCuculiphilus fasciatusArdeiphilus trochioxusOdoriphila clayae/phoeniculiPsittacomenopon poicephalusColpocephalum zebraComatomenopon elbeli/elongatum

Gruimenopon longumHoazineus armiferusMeromenopon meropisTrinoton anserinumMenacanthus stramineusColimenopon urocoliusSomaphantus lusiusAmyrsidea ventralisMenopon gallinaeNumidicola antennatus

B

L

M B

C

D

A

Liposcelis bostrychophilus

Marshall, 200344 genera, 147 char.(Morphology)

Psocoptera

Menacanthus /Menopon - complex

Ricinidae

Laemobothriidae

Boopiidae

Austromenopon- complex

Dennyus- complex

Colpocephalum- complex

• Collected Eckfeld maar nearEifel, Germany

• Middle Eocene (Middle Lutetian,44.3±0.4 Ma)

• First fossil louse

• Completes ordinal representationof fossil insects?

• Excellent preservation

• Phylogenetic affinities withmodern feather lice

• Parasite of Anseriformes orCharadriiformes

• Crown group position• Points to a long coevolutionary

history with birds• Suggests lice are very old• Useful molecular calibration point

First Fossil Louse (Amblycera: Menoponidae)

Wappler, Smith &Dalgleish , In prep.

Ischnoceran FamiliesGenusFamilySuborder

3

532

3,060

1,344

MammalsBirds

Amblycera

Ischnocera

Rhyn…

Anoplura

Ischnocera

TrichodectidaeHeptapsogasteridae

“Philopteridae”

Hopkins & Clay, 1951

cf. Eicher, 196121 families

Smith, 2000Generic level phylogeny ofGoniodidae & Heptapsogasteridae

(Morphology)

Heptapsogasteridae

Goniodidae

Avian Ischnoceran Phylogeny

Columbicola columbae

Austrogoniodes waterstoni

Harrisoniella hopkinsiHaffneria grandis

Paraclisis diomedeaPerineus nigrolimbatusHalipeurus pelagicusPhiloceanus garrodiaePseudonirmus gurltiNaubates fuliginosusPelmatocerandra setosaArdeicola smithersiAnaticola crassicornisPectinopygus bassaniPectinopygus sulaeAquanirmus australisIbidoecus plataleaeTrabeculus schillingiCraspedonirmus colymbinusQuadraceps coenocoryphaeDocophoroides brevisSaemundssonia desolataSaemundssonia haematopiPodargoecus strigoidesAlcedoecus delphaxCraspedorrhynchus platystomusStrigiphilus vapidusSturnidoecus sturniPhilopterus ornatusVernoniella guimarãesiAcidoproctus rostratusAcidoproctus hilliRallicola lugensOsculotes curtusOsculotes macropodaColoceras damicorneCampanulotes bidentatusGoniocotes gallinaeGoniodes pavonisGoniodes kéleriChelopistes guttatus

Strongylocotes angulocapitisDiscocorpus c. cephalosus

Splendoroffula ruwenzorornisOxylipeurus dentatusCuclotogaster madagascariensisLipeurus caponisDegeeriella rufaUpupicola upupaeLagopoecus affinisSyrrhaptoecus falcatusNeopsittaconirmus borgioliiParagoniocotes rotundusBrüelia semiannulataFalcolipeurus affulgeusArcholipeurus nandu

Felicola (F.) viverriculaeGeomydoecus (G.) heaneyi

Trichodectes (T.) melisBovicola limbatusDamalinia (D.) crenelata

Smith, 2001Generic level phylogenyApprox. 1/3rd of all ischnoceran genera

51 genera, 138 characters(Morphology)

Philoceanus-complex

Goniodidae

Degeeriella-complex

Trichodectidae

Lipeurus-complex

Heptap.

Saemundssonia-complex

Philopterus-complex

Multiple clades of liceon the same host

A. Circumfasciate Head

B. Non-Circumfasciate Head

N I N II N III Adult

C. Wing Lice D. Body Lice E. Head Lice

Characters Defining Major Ischnoceran Clades

Philoceanus-complex

Goniodidae

Degeeriella-complex

Trichodectidae

Lipeurus-complex

Heptap.

Saemundssonia-complex

Philopterus-complex

WingLice

WingLice

CI: 0.5RI: 0.95

HeadLice

BodyLiceCircumfasciate

Non-circumfasciate

CI: 1.0RI: 1.0

Characters Defining Major Ischnoceran Clades

Multiple cladesoccupy differenthost niches

• Incongruent with molecular data• Morphological convergence• Molecular data suggests many of

these pairs are more closely related

• Character displacement?

• Sympatric speciation?

Problems with Morphology!Smith et al, 2003

Rhynchophthirina: HaematomyzidaeGenusFamilySuborder

3

532

3,060

1,344

Amblycera

Ischnocera

Anoplura

Rhynchophthirina

Rhynchophthirina: Haematomyzidae• Just 3 species in 1 family• Present on elephants, wart hog & bush

pig (perhaps also mammoths?)• Monophyly & sister placement with

Anoplura well supported• Chewing lice (have outwardly opposing

mandibles)• Pool blood feeders (telmophages) cf

vessel feeding Anoplura• Spread via shared use of watering holes?• Possible cryptic species?

(morphologically identical species onAfrican & Asian elephants)

• Specimens hard to come by!

Haematomyzus elephantisHaematomyzus elephantisex Asian / African elephantex Asian / African elephant

Haematomyzus elephantisHaematomyzus hopkinsiHaematomyzus porci

Part of Smith, in prep.51 genera, 76 species184 characters(Morphology)

3

532

3,060

1,344

Amblycera

Ischnocera

Rhyn…

Anoplura

Anopluran Families

Anoplura At least 15distinct families

Anoplura:familial phylogeny

• 541spp. in 15 families• Patchily distributed across

mammals

Based on Kim, 198847 genera, 39 characters(Morphology)

• Monophyly of all 15 familiessupported

• 3 basal clades• Many lice of host groups

(including Primates) paraphyletic

Kim’s datasetre-analysed:

Kim, 1988 reanalysedStrict Consensus

• Length=219 (CI=0.26, RI=0.62)

OUTGROUP

“Polyplacidae” 1



Hoplopleuridae

“Polyplacidae” 4-12

Echinophthiriidae

Enderleinellidae

“Linognathidae” 1

Neolinognathidae


Pedicinidae

Pthiridae


Ratemiidae

Hamophthiriidae

Hybophthiridae

Pediculidae

Pecaroecidae

Haematopinidae

Microthoraciidae




“Polyplacidae” 4-12




• Paraphyly of 2 of the 5 familiesthat were not monogenic

• Only 3 other nodes arecompatible with Kim’s tree

• Just 1 node agrees with Kim’soriginal tree!

Haematomyzus elephantisHaematomyzus hopkinsi

Pthirus gorillaePthirus pubisPediculus schaeffiPediculus humanusPediculus mjobergiPediculus humanus var. americanusScipio aulacodiScipio tripedatusNeolinognathus sp.Neolinognathus elephantuliSolenopotes muntiacusProlinognathus aethiopicusProlinognathus ferrisiLinognathus setosusLinognathus aepycerusLinognathus africanusLatagophthirus rauschiEchinophthirius horridusLepidophthirus macrorhiniAntarctophthirus callorhiniAntarctophthirus microchirProechinophthirus fluctusProechinophthirus zumptiJohnsonpthirus chlorotalpaeJohnsonpthirus heliosciuriLemurphthirus galagusLemurphthirus stigmosusLinognathoides laeviusculusSathrax durusNeohaematopinus callosciuriNeohaematopinus inornatusAbrocomaphthirus chilensisCtenophthirus cercomydisProenderleinellus calvaPolyplax abyssinicaPolyplax spinulosaDocophthirus acinetusEulinognathus denticulatusHaemodipsus leporisHaemodipsus lyriocephalusLagidiophthirus parvusFahrenholzia ferrisiFahrenholzia pinnataGaleophthirus caviaePhthirpediculus brygooiPhthirunculus sumatranusAtopophthirus emersoniMicrophthirus uncinatusEnderleinellus osborniWerneckia nigeriensisHoplopleura acanthopusParadoxophthirus emarginataHoplopleura pacificaHoplopleura irritansPterophthirus audaxPterophthirus splendidaAncistroplax crociduraeHaematopinoides squamosusTyphlomyophthirus bifoliatusSchizophthirus graphiuriSchizophthirus pleurophaeus

Haematomyzus porciMicrothoracius cameliRatemia squamulataHamophthirius galeopitheciHybophthirus notophallusHaematopinus asiniHaematopinus suisPecaroecus javaliiPedicinus ferrisiPedicinus patas

Anoplura:phylogeny

Haematomyzidae

Linognathidae

“Polyplacidae” 1Neolinognathidae

Enderleinellidae

Hoplopleuridae



Echinophthiriidae

MicrothoraciidaeRatemiidaeHamophthiriidaeHybophthiridae

HaematopinidaePecaroecidae

PedicinidaePthiridae

Pediculidae

Smith, in prep.51 genera, 76 species184 characters(Morphology)

• Why are rodents so lousy?

• Why are other mammalsrelatively louse free cf. birds

• Good evidence forcophylogeny in some clades

• Comparative studies underwayusing these trees

Louse Molecular Studies

Nuclear

- EF1-α (350-450bp)- 18S rRNA (1500+bp)- Wingless, PEPCK, DDC

• Hard to amplify• Multiple copies• RNA alignment issues• Utility varies between genes &

taxa

Mitochondrial

- COI (400-1500bp)- 12S rRNA (450bp)- Cyt. B (450bp)- ND4 (500bp)

• Easy to amplify• Limited phylogenetic utility

outside families• Often restricted to use within

closely related genera

Need New Genes!

Lice

all sitespos. 1pos. 2pos. 3



11

0.12129.9

11

0.15169.78

11

0.000144.79

0.0180.0130.0160.41

0.0160.0390.017

0.80.250.20

0.00012.49

55.676.97.5

316.861.525.68.8

212.24.15.01.018.0

Physconelloidinae

Columbicola

aphids

DataSet

Sites Ratio(COI:EF1α)COI EF1-α

Rate Parameter

Relative Substitution Rate(Mit. COI vs nuc. EF1-α)

Louse 12SSecondary

Structure

mit. 12S rRNAColumbicola columbae

Austrogoniodes cristati

Naubates prioni

Halipeurus pelagicus

Naubates harrisoni

Rallicola columbiana

Austrophilopterussubsimilis

Austrogoniodes cristati

Austrogoniodes waterstoni

Oxylipeurus chiniri

Campanulotes bidentatus

Austrophilopterussubsimilis

Page et al, 2002

0

20

40

60

80

100

120

Length

35 40 45 50 55 60 65 70 75 800123456789

10

Length

020406080

100120140160180200

Length

15 20 2530 35 40 45 50 55 60 65 70 75 80 850

2

4

6

8

10

12

14

Length

Segment 3: 38'-39' Segment 5: 34'-33'

0

20

40

60

80

100

120

140

160

Length

5 10 15 20 25 30 35 40 450

2

4

6

8

10

12

Length

Segment 2: 36-38Segment 1: 33-33'

0

20

40

60

80

100

120

140

160

Length

insects

2102202302402502602702802903003103200

1

2

3

4

5

6

7

Length

lice

12S LengthVariation

Insects vs.LicePage et al, 2002

cox1 cox2

atp8

atp6 cox3

nad3 nad5 nad4

nad4

L

nad6 cob nad2 rrnL rrnS nad2

L2 KD G HARNS1EF TP VS2 L1IQM WCY

cox1 cox3 nad1 cob nad4

nad4

L

nad2

nad3nad5 cox2 rrnL rrnS atp6

atp8

nad6

S2 L2 KNAY QV PTDIFS1CMWERII GL1

Gene arrangement of Heterodoxus macropus (Amblycera)

Inferred gene arrangement of ancestral insect

Adapted fromAdapted from Shao Shao, Campbell & Barker, 2001, Campbell & Barker, 2001

Louse Mitochondrial Genome

Louse Molecular Studies

Nuclear

- EF1-α (350-450bp)- 18S rRNA (1500+bp)- Wingless, PEPCK, DDC

• Hard to amplify• Multiple copies• RNA alignment issues• Utility varies between genes &

taxa

Mitochondrial

- COI (400-1500bp)- 12S rRNA (450bp)- Cyt. B (450bp)- ND4 (500bp)

• Easy to amplify• Limited phylogenetic utility

outside families• Often restricted to use within

closely related genera

Need New Genes!

ISCHNOCERA

AMBLYCERA

ANOPLURA

RHYN… Louse NuclearGenes

CruickshankCruickshank et al, 2001 et al, 2001

• 350 bp single copy EF1-α• Limited phylogenetic utility

above families

What Not to do!

Phylogenetic Utility 18S rRNA

Smith et al, 2003

• Much recent progress on basal phthirapteran phylogeny• Monophyly of suborders well supported• Complex pattern of host-louse relationships• Distinctive molecular evolution in lice• Incongruence between molecules & morphology

Ongoing Questions…

• To what extent do lice cospeciate with their hosts?• Why are some hosts more lousy than others?• How old are the major louse clades?• Are Phthiraptera truly paraphyletic?

Conclusions…

Acknowledgements

CollaborationStephen BarkerDale Clayton Tom FordKevin JohnsonRod PageIsabel MarshallDavid Reed

FundingWellcome Trust, UKNatural Environment Research Council, UK

0.60.50.40.30.20.100

100

200

300

ML Corrected Divergences(EF1-α 3rd Positions)

ML

Cor

rect

ed D

iver

genc

es(C

OI 3

rd P

ositi

ons)

Slopeapproximatesrelative rate

Around 1000:1 !!!

Pairwise ML Corrected Distances for 3rd Sites(Nuclear EF1-α against Mit. COI)

Johnson et al, 2003

18S – not a panacea for basal louse Phylogeny!

Unc

orre

cted

NJ

Bay

esia

n

0.10.1

AnopluraAnoplura

Ischnocera

Ischnocera

Amblycera

Amblycera

Liposcelis sp.

Liposcelis sp.Trogiomorph Psocoptera Trogiomorph Psocoptera

2003.v smith.dresden

Technology