Lousy Phylogenies:

1
Lousy Phylogenies
Phthiraptera systematics and the antiquity of lice
Vincent S. Smith
Division of Environmental and Evolutionary
Biology Institute of Biomedical and Life
Sciences Graham Kerr Building University of
Glasgow Scotland, UK
2
Lice (Phthiraptera) biology

1. Cophylogeny, cospeciation coevolution

3
Lice (Phthiraptera) taxonomy
- 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
4
Lice (Phthiraptera) phylogeny
Chewing Lice Mallophaga
Sucking Lice (Anoplura)
5
Genus
Family
Suborder
532
3
3,060
1,344
6
Genus
Family
Suborder
Anoplura
Phthiraptera Phylogenetic resolution, 2003
532
3
Rhyn
3,060
Ischnocera
1,350
Amblycera
7
1996
Genus
Family
Suborder
Anoplura
532
3
Rhyn
3,060
Ischnocera
1,344
Amblycera
8
Amblyceran Families
Menoponidae
Boopiidae
85
Laemobothriidae
Ricinidae
Trimenoponidae
Protogyropinae
84
Gyropinae
Analysed by Marshall, 2003 from Clay, 1970
(Morphology)
Grilicolinae
Liposcelidae (Psocoptera)

9
Avian Amblycera
Liposcelis bostrychophilus
Trochiloecetes rupununi
Ricinus fringillae
Trochiliphagus abdominalis
Laemobothrion maximum
L
Therodoxus oweni
Boopia tarsata
Paraheterodoxus insignis
B
Latumcephalum lesouefi/macropus
Paraboopia flava
Rediella mirabilis
Actornithophilus uniseriatus
Plegadiphilus threskiornis
Chapinia robusta
Bonomiella columbae
Pseudomenopon pilosum
Ancistrona vagelli
Dennyus hirundinis
A
Myrsidea victrix
Holomenopon brevithoracicum
Eidmanniella pellucida
Machaerilaemus laticorpus/latifrons
Austromenopon crocatum
Neomenopon pteroclurus
B
M
Hohorstiella lata
Osborniella crotophagae
Eomenopon denticulatum
Piagetiella bursaepelecani
Ciconiphilus quadripustulatus
Cuculiphilus fasciatus
Ardeiphilus trochioxus
Odoriphila clayae/phoeniculi
Psittacomenopon poicephalus
C
Colpocephalum zebra
Comatomenopon elbeli/elongatum
Gruimenopon longum
Hoazineus armiferus
Meromenopon meropis
Marshall, 2003
Trinoton anserinum
Menacanthus stramineus
44 genera, 147 char. (Morphology)
Colimenopon urocolius
Somaphantus lusius
Amyrsidea ventralis
D
Menopon gallinae
Numidicola antennatus

10
First Fossil Louse (Amblycera Menoponidae)
Wappler, Smith Dalgleish , In prep.
11
Ischnoceran Families
Genus
Family
Suborder
Anoplura
532
3
Rhyn
3,060
Ischnocera
1,344
cf. Eicher, 1961 21 families
Amblycera
12
Avian Ischnoceran Phylogeny
Multiple clades of lice on the same host
13
Characters Defining Major Ischnoceran Clades
14
Characters Defining Major Ischnoceran Clades
Multiple clades occupy different host niches
15
Rhynchophthirina Haematomyzidae
Genus
Family
Suborder
Anoplura
532
3
3,060
Ischnocera
1,344
Amblycera
16
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 on African Asian elephants)
• Specimens hard to come by!

Haematomyzus elephantis
ex Asian / African elephant
17
Anopluran Families
Anoplura
532
3
Rhyn
3,060
Ischnocera
1,344
Amblycera
18

• 541spp. in 15 families
• Patchily distributed across mammals

• Monophyly of all 15 families supported

19
Kims dataset re-analysed
OUTGROUP
Polyplacidae 1
Polyplacidae 2
Polyplacidae 3
Hoplopleuridae

• Length219 (CI0.26, RI0.62)

Polyplacidae 4-12
Echinophthiriidae
Enderleinellidae
Linognathidae 1
Neolinognathidae

• Only 3 other nodes are compatible with Kims tree

Linognathidae 2
Pedicinidae
Pthiridae
Polyplacidae 13
Ratemiidae
Hamophthiriidae
Hybophthiridae
Pediculidae
Pecaroecidae
Haematopinidae
Microthoraciidae
20
Haematomyzus elephantis
Haematomyzus hopkinsi
Haematomyzus porci
Microthoracius cameli
Ratemia squamulata
Hamophthirius galeopitheci
Hybophthirus notophallus
Haematopinus asini
Haematopinus suis
Pecaroecus javalii
Pedicinus ferrisi
Pedicinus patas
Pthirus gorillae
Pthirus pubis
Pediculus schaeffi
Pediculus humanus
Pediculus mjobergi
Pediculus humanus var. americanus
Scipio aulacodi
Scipio tripedatus
Neolinognathus sp.
Neolinognathus elephantuli
Solenopotes muntiacus
Prolinognathus aethiopicus
Prolinognathus ferrisi
Linognathus setosus
Linognathus aepycerus
Linognathus africanus
Latagophthirus rauschi
Echinophthirius horridus
Lepidophthirus macrorhini
Antarctophthirus callorhini
Antarctophthirus microchir
Proechinophthirus fluctus
Proechinophthirus zumpti
Johnsonpthirus chlorotalpae
Johnsonpthirus heliosciuri
Lemurphthirus galagus
Lemurphthirus stigmosus
Linognathoides laeviusculus
Sathrax durus
Neohaematopinus callosciuri

• Why are rodents so lousy?
• Why are other mammals relatively louse free cf.
  birds
• Good evidence for cophylogeny in some clades

Neohaematopinus inornatus
Abrocomaphthirus chilensis
Ctenophthirus cercomydis
Proenderleinellus calva
Polyplax abyssinica
Polyplax spinulosa
Docophthirus acinetus
Eulinognathus denticulatus
Haemodipsus leporis
Haemodipsus lyriocephalus
Lagidiophthirus parvus
Fahrenholzia ferrisi
Fahrenholzia pinnata
Galeophthirus caviae
Phthirpediculus brygooi
Phthirunculus sumatranus
Atopophthirus emersoni
Microphthirus uncinatus
Enderleinellus osborni
Werneckia nigeriensis
Hoplopleura acanthopus

• Comparative studies underway using these trees

Paradoxophthirus emarginata
Hoplopleura pacifica
Hoplopleura irritans
Pterophthirus audax
Pterophthirus splendida
Ancistroplax crocidurae
Haematopinoides squamosus
Typhlomyophthirus bifoliatus
Schizophthirus graphiuri
Schizophthirus pleurophaeus
21
Louse Molecular Studies
Nuclear
Mitochondrial
- EF1-a (350-450bp) - 18S rRNA (1500bp) -
Wingless, PEPCK, DDC
- COI (400-1500bp) - 12S rRNA (450bp) - Cyt. B
(450bp) - ND4 (500bp)

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

• Easy to amplify
• Limited phylogenetic utility outside families
• Often restricted to use within closely related
  genera

Need New Genes!
22
Relative Substitution Rate
(Mit. COI vs nuc. EF1-a)
Data Set
Ratio (COIEF1a)
Sites
Physconelloidinae
Lice
Columbicola
aphids
23
Louse 12S Secondary Structure
mit. 12S rRNA
Page et al, 2002
24
12S Length Variation
Segment 5 34'-33'
120
100
80
60
40
20
0
Length
10
9
8
7
6
5
4
3
2
1
Page et al, 2002
0
35
40
45
50
55
60
65
70
75
80
Length
25
Louse Mitochondrial Genome
Inferred gene arrangement of ancestral insect
Adapted from Shao, Campbell Barker, 2001
26
Louse Molecular Studies
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!
27
Louse Nuclear Genes
RHYN
ANOPLURA
What Not to do!

• 350 bp single copy EF1-a
• Limited phylogenetic utility above families

ISCHNOCERA
AMBLYCERA
Cruickshank et al, 2001
28
Phylogenetic Utility 18S rRNA
Smith et al, 2003
29
Conclusions

• 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

30
Acknowledgements
Collaboration Stephen Barker Dale Clayton Tom
Ford Kevin Johnson Rod Page Isabel Marshall David
Reed
Funding Wellcome Trust, UK Natural Environment
Research Council, UK
31
Pairwise ML Corrected Distances for 3rd Sites
(Nuclear EF1-a against Mit. COI)
300
200
ML Corrected Divergences
(COI 3rd Positions)
100
0
0.6
0.5
0.4
0.3
0.2
0.1
0
ML Corrected Divergences
(EF1-a 3rd Positions)
Johnson et al, 2003
32
18S not a panacea for basal louse Phylogeny!
Trogiomorph Psocoptera
Trogiomorph Psocoptera
Liposcelis sp.
Amblycera
Ischnocera
Liposcelis sp.
Ischnocera
Amblycera
Uncorrected NJ
Anoplura
Anoplura
Bayesian
0.1
0.1