Additional file 1: s of reptiles Summary of detailed categorization by bile Class Description Number of reptile species I Only bile 0 II Mixture of bile and bile III Mixture of bile and bile 0 IV Only bile 67 V Mixture of bile and bile 5 VI All bile 146 Total 219 Mostly 5 (A/B trans, allo ) 35 2 (Siebenrock s -neck and California desert tortoise) Summary of diet sorted by primary bile Diet Total # CA CDCA Other bile bile bile alcohol Complex profile* Carnivores 183 61.7% 0% 24.6% 24.6% 0% 9.8% Herbivores 15 33.3% 0% 0% 66.7% 0% 6.7% Omnivores 17 11.8% 0% 0% 88.2% 0% 5.9% *Complex profile defined as: (1) bile II, III, or V; and/or (2) presence of 3 or more bile s in biliary bile, each at 10% or more Abbreviations and bile names used: CA (cholic ; 3,7,12 -trihydroxy--cholan-24-oic ) common bile CDCA (chenodeoxycholic ; 3,7 -dihydroxy--cholan-24-oic ) common stem bile DCA (deoxycholic ; 3,12 -dihydroxy-5 -cholan-24-oic ) common secondary bile Allo- - -bile (e.g., alloca or allocdca) LCA (lithocholic ; 3 -hydroxy-5 -cholan-24-oic ) - common (often toxic) secondary bile PythoCA (pythocholic ; 3,12,16-5 -cholan-24-oic ) unusual bile lacking 7 -OH; found in some s SteroCA (sterocholic ; 3,7,12,22-tetrahydroxy--cholestan-27-oic ) 22-hydroxylated bile found in s SteroCDCA (sterochenodeoxycholic ; 3,7,22-trihydroxy--cholestan-27-oic ) bile found in s Varanic (3,7,12α,24R-tetrahydroxy--cholestan-27-oic ) 24R-hydroxylated bile typical of varanid lizards
Additional file 1 (cont.), p. 2 Conjugation Major bile s Methodology Comments REPTILES TESTUDINES Carettochelyididae Carettochelys insculpta Plateless IV X Taurine Very complex mixture of bile All s analyzed use bile conjugated with taurine and use rather unique 22-hydroxylation (sterochenodeoxycholic and sterocholic ) Very low levels of 12α-hydroxylation Chelidae Chelodina novaeguineae Chelodina siebenrocki Chelus fimbriatus New Guinea -neck Siebenrock s -neck Mata Mata IV X Taurine SteroCA, sterocdca II X X X Taurine Complex mixture of bile (70%), bile (22%), and bile (8%) ESI/MS/MS IV X Taurine SteroCA, sterocdca Cheloniidae Caretta caretta Loggerhead IV X Taurine SteroCA, sterocdca, sterodca Chelonia midas Green IV X Taurine SteroCA, sterocdca, sterodca Eretmochelys imbricata Lepidochelys olivacea Hawks bill Olive Ridley IV X Taurine SteroCA, sterocdca, sterodca IV X Taurine SteroCA, sterocdca, sterodca Approximately 10% unidentified bile likely representing complex stereoisomers Approximately 10% unidentified bile likely representing complex stereoisomers Approximately 6% unidentified bile likely representing complex stereoisomers Approximately 20% unidentified bile likely representing complex stereoisomers
Additional file 1 (cont.), p. 3 Conjugation Major bile s Methodology Comments Chelydridae Macrochelys temminckii Alligator snapping IV X Taurine SteroCA, sterocdca, sterodca SteroDCA may result from enterohepatic circulation Dermochelyidae Dermochelys coriacea Leatherback IV X Taurine SteroCDCA, steroca Approximately 40% unidentified complex mixture of bile Emydidae 1-Hydroxylated bile also found in this family. Chrysemys picta Painted IV X Taurine SteroCA, sterocdca Glyptemys Wood IV X Taurine SteroCA, sterocdca insculpta Graptemys Map IV X Taurine SteroCA, sterocdca geographica Malaclemys Diamondback IV X Taurine SteroCA, sterocdca terrapin terrapin Pseudemys Common IV X Taurine SteroCA, sterocdca concinna cooter Trachemys scripta Red-eared IV X Taurine SteroCA, sterocdca Geoemydidae Chinemys reevesii Cuoro galbinifrons Cuora trifasciata Cyclemys tcheponensis Reeves IV X Taurine SteroCA, sterocdca Indochinese box Chinese threestriped Stripe-necked leaf IV X Taurine SteroCA, sterocdca IV X Taurine SteroCA, sterocdca IV X Taurine SteroCA, sterocdca Podocnemidae Podocnemis expansa South American river IV X Taurine SteroCA, sterocdca
Additional file 1 (cont.), p. 4 Conjugation Major bile s Methodology Comments Testudinidae Chelonoidis Yellow-footed denticulata tortoise Geochelone nigra Geochelone sulcata Gopherus agassizii Manouria emys Manouria impressa Psammobates pardalis Pyxis arachnoides Testudo horsfieldii Galapagas tortoise African spurred tortoise California desert tortoise Asian brown tortoise Impressed tortoise Leopard tortoise Madagascar spider tortoise Central Asian tortoise IV X Taurine 25R and 25S substituted steroca and sterocdca IV X Taurine 25R and 25S substituted steroca and sterocdca IV X Taurine 25R and 25S substituted steroca and sterocdca II X X Taurine, sulfate bile ; C 26,, and C 28 bile alcohol sulfates (30% of total bile s) IV X Taurine 25R and 25S substituted steroca and sterocdca IV X Taurine 25R and 25S substituted steroca and sterocdca V X X Taurine CA, CDCA, 25R and 25S substituted steroca and sterocdca V X X Taurine CA, CDCA, bile IV X Taurine 25R and 25S substituted steroca and sterocdca ESI/MS/MS LC/MS LC/MS Highest bile alcohol sulfate content in biliary bile for any reptile examined. C 26 and C 28 bile alcohol sulfates are very rare in vertebrates other than amphibians. Trionychidae Chitra indica Pelochelys bibronii Pelodiscus sinensis Narrow-head soft shell Bibron s soft shell Chinese soft shell IV X Taurine SteroCA IV X Taurine SteroCA IV X Taurine SteroCA
Additional file 1 (cont.), p. 5 Conjugation Major bile s Methodology Comments SPHENODONTIA Sphenodon punctatus Tuatara IV X Taurine 24R-hydroxylated bile LIZARDS, GECKOES, CHAMELEONS Agamidae Agama agama Red-headed rock agama VI X Taurine AlloCA Chlamydosaurus Frilled lizard VI / X Taurine AlloCA, 25% CA kingii Laudakia stellio Starred agama VI X Taurine AlloCA Physignathus Chinese water VI X Taurine AlloCA, CA cocincinus dragon Physignathus Leseur s water VI X Taurine AlloCA, CA lesueurii dragon Pogona barbata Eastern VI X Taurine AlloCA, CA bearded dragon Pogona vitticeps Inland bearded VI X Taurine AlloCA, CA, GC/MS dragon Uromastyx acanthinura North African spiny-tailed VI X Taurine Mostly alloca, also allodca Uromastyx aegyptia microlepis Uromastyx benti Uromastyx hardwicki Uromastyx ocellata Uromastyx thomasi lizard Egyptian spiny-tailed lizard Benti s spinytailed lizard Indian spinytailed lizard Ornate spinytailed lizard Thomas spiny-tailed lizard VI X Taurine Mostly alloca, also allodca VI X Taurine Mostly alloca, also allodca VI X Taurine Mostly alloca, also allodca VI X Taurine Mostly alloca, also allodca VI X Taurine Mostly alloca, also allodca
Additional file 1 (cont.), p. 6 Conjugation Major bile s Methodology Comments Anguidae Elgaria multicarinata Pseudopus apodus California alligator lizard Scheltopusik glass lizard V X X Taurine Varanic, 6% bile V X X Taurine Varanic, 8% bile Chamaeleonidae Brookesia perarmata Calumma parsonii Chamaeleo calyptratus Chamaeleo gracilis Chamaeleo jacksonii Chamaeleo melleri Chamaeleo montium Chamaeleo quadricornis Furcifer oustaleti Furcifer pardalis Rampholeon spectrum Rieppeleon brevicaudatus Armored Parson s Veiled Graceful Jackson s Meller s Sailfin Four-horned Oustalet s Panther Leaf Stump-tail VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca VI X Taurine Mostly alloca Cordylidae Cordylus warreni Warren s girdle-tail VI X Taurine alloca + complex mixture of other bile
Additional file 1 (cont.), p. 7 Conjugation Major bile s Methodology Comments Gekkonidae Cyrtodactylus Malayan benttoed VI X Taurine CA, CDCA pulchellus gecko Gekko gecko Tokay gecko VI X Taurine alloca, allocdca Hemitheconyx Fat-tailed VI / X Taurine CA, CDCA caudicinctus Phelsuma flavigularis Ptychozoon kuhli Helodermatidae Heloderma suspectum gecko Yellowthroated day gecko Gliding gecko VI / VI X Taurine CA, CDCA X Taurine CA, CDCA, alloca Gila monster IV X Taurine Mostly varanic Iguanidae Anolis Green anole VI / caroliensis Anolis equestris Knight anole VI / Anolis grahami Jamaican anole VI / Brachylophus Hybrid Fiji VI / fasciatus Island green iguana Corytophanes Casque-headed VI / cristatus iguana Cyclura carinata Turks and VI / Caicos iguana Cyclura cornuta Rhinoceros VI / cornuta iguana Sauromalus ater Chuckwalla VI / X Taurine Mostly alloca X Taurine Equal mixture of alloca and CA X Taurine Mostly alloca, only 6% bile X Taurine Mostly alloca X Taurine AlloCA, allocdca, only 15% bile X Taurine Mostly alloca, only 8% bile X Taurine Mostly alloca X Taurine Mostly alloca, only 8% bile Lacertidae
Additional file 1 (cont.), p. 8 Conjugation Major bile s Methodology Comments Lacerta viridis Emeralda lacerta VI / X Taurine Mostly CA, some alloca Scincidae Egernia depressa Trachylepis affinis Pygmy spinytailed skink Blotched bluetailed skink IV X Taurine >97% bile V X X Taurine bile, ~35% bile ESI/MS/MS Also traces of unusual bile alcohol sulfates Teiidae Ameiva ameiva Blue ameiva VI X Taurine Mostly CA Cnemidophorus St. Lucia VI X Taurine Mostly CA vanzoi whiptail Dracaena Caiman lizard VI X Taurine Mostly CA guianensis Tupinambis sp. Tegu VI X Taurine Mostly CA Varanidae Varanus White-throated IV X Taurine Mostly varanic albigularis monitor Varanus Black-tree IV X Taurine Mostly varanic beccarii monitor Varanus Bengal monitor IV X Taurine Mostly varanic, ESI/MS/MS bengalensis Varanus Savannah IV X Taurine Mostly varanic exanthematicus monitor Varanus Savannah IV X Taurine Mostly varanic exanthematicus monitor Varanus gouldi Gould s IV X Taurine Mostly varanic monitor Varanus indicus Mangroe IV X Taurine Mostly varanic monitor Varanus Peach-throated IV X Taurine Mostly varanic jobiensis monitor Varanus Komodo IV X Taurine Mostly varanic komodoensis monitor Varanus Emerald IV X Taurine Mostly varanic
Additional file 1 (cont.), p. 9 Conjugation Major bile s Methodology Comments kordensis monitor Varanus melinus Quince IV X Taurine Mostly varanic monitor Varanus Nile monitor IV X Taurine Mostly varanic niloticus Varanus Gray s monitor IV X Taurine Mostly varanic olivaceus Varanus New Guinea IV X Taurine Mostly varanic panoptes monitor Varanus Rough-neck IV X Taurine Mostly varanic rudicollis monitor Varanus Two-banded IV X Taurine Mostly varanic salvator monitor Varanus Timor monitor IV X Taurine Mostly varanic timorensis Varanus varius Lace monitor IV X Taurine Mostly varanic SNAKES Acrochordidae Acrocordus javanicus Acrocordus granulates Elephant trunk File VI / VI X Taurine CA Acrochordids no longer show 16αhydroxylation X Taurine CA, DCA, alloca Boidae Acrantophis madagascariensis Boa constrictor imperator Boa constrictor occidentalis Boa constrictor ortonii Calabaria reinhardtii Candoia bibroni australis Madagascar VI X Taurine Mostly pythoca ground boa Red-tail boa VI X Taurine Mostly pythoca Argentine boa VI X Taurine Mostly pythoca Red-tailed boa VI X Taurine Mostly pythoca Calabar ground python Solomon Islands boa VI X Taurine Mostly pythoca VI X Taurine Mostly pythoca
Additional file 1 (cont.), p. 10 Conjugation Major bile s Methodology Comments Corallus caninus Eryx johnii Lichanura australis roseofusca Emerald tree boa Brown sand boa Coastal rosy boa VI X Taurine Mostly pythoca VI X Taurine Mostly pythoca VI X Taurine Mostly pythoca Colubridae Ahaetulla nasuta Dispholidus typhus Drymarchon corais Elaphe quatuorlineata Euprepiophis mandarina Gongylosoma longicauda Lampropeltis getulus Lampropeltis triangulum Macrocalamus lateralis Masticophis flagellum testaceus Masticophis taeniatus Orthriophis moellendorfi Green tree VI / X Taurine Mostly CA, 25% alloca Boomslang VI X Taurine Mostly CA Indigo VI X Taurine Mostly CA Four-lined rat Mandarin rat Striped gopher Speckled king VI X Taurine Mostly CA VI X Taurine Mostly CA VI X Taurine Mostly CA, minor fraction of 22 and bile VI X Taurine Mostly CA, minor fraction of 22 bile Milk VI X Taurine Mostly CA, minor fraction of 22 bile Malayan mountain reed Western coachwhip Striped whip Red-head rat AlloCA may be from diet VI X Taurine Mostly CA Minor fractions of bile alcohol sulfates identified VI X Taurine Mostly CA VI X Taurine Mostly CA VI X Taurine Mostly CA
Additional file 1 (cont.), p. 11 Conjugation Major bile s Methodology Comments Pituophis deppei Queretaro VI X Taurine Mostly CA jani gopher Ptyas mucosus Dhaman VI X Taurine Mostly CA Stegonotus Northern VI X Taurine Mostly CA modestus ground Telescopus African tiger VI X Taurine Mostly CA semiannulatus Thelotornis Twig VI X Taurine Mostly CA kirtlandii Zamenis longissimus Aesculapian VI X Taurine Mostly CA Cylindrophiidae Cylindrophis maculatus Cylindrophis rufus Blotched pipe VI X Taurine PythoCA, 7α-OHpythoCA Pipe VI X Taurine PythoCA, 7α-OHpythoCA Dipsadidae Farancia abacura Mud VI X Taurine -CA, CA, and DCA Elapidae Acanthophis antarcticus Bungarus fasciatus Bungarus flaviceps Calliophis bivirgata Dendroaspis polylepis Laticauda colubrine Death adder VI X Taurine Mostly CA Banded krait VI / Red-head trait VI / Blue VI / Malaysian coral South African mamba Yellow-lipped sea X Taurine CA, minor fraction of alloca X Taurine CA, minor fraction of alloca X Taurine CA, minor fraction of alloca VI X Taurine Mixture of and C 23 bile, some with VI X Taurine Mostly CA
Additional file 1 (cont.), p. 12 Conjugation Major bile s Methodology Comments Laticauda semifasciata Naja haje Naja pallida Notechis scutatus Ophiophagus hannah Pelamis platurus Pseudechis colletti Erabu sea VI X Taurine Mostly CA Egyptian VI / X Taurine Mostly CA, LC/MS/MS banded cobra Red-spitting VI / X Taurine CA, minor fraction of cobra alloca Australian tiger VI X Taurine Mostly CA King cobra VI / X Taurine CA, minor fraction of alloca Yellow-bellied VI X Taurine Mostly CA sea Collett s black VI X Taurine Mostly CA Homalopsidae Enhydris plumbea Homalopsis buccata Plumbeous water Puff-faced water VI X Taurine -CA, CA VI X Taurine -CA, CA Lamprophiidae Lycodonomorphus Red wolf VI X Taurine Mostly CA rufulus Psammophis Indo-Chinese VI X Taurine Mostly CA condanarus sand Pseudaspis cana Mole VI / X Taurine CA, minor fraction of alloca Natricidae Nerodia erythrogaster Nerodia fasciata fasciata Opisthotropis lateralis Storeria occipitomaculata Plain-bellied water VI X Taurine Mostly CA Banded water VI X Taurine Mostly CA Bicolored VI X Taurine Mostly CA stream Red-bellied VI X Taurine Mostly CA
Additional file 1 (cont.), p. 13 Conjugation Major bile s Methodology Comments Pareatidae Pareas margaritophorus Asthenodipsas vertebralis White-spotted slug Mountain slug VI X Taurine Mostly CA VI X Taurine Mostly CA Pythonidae Aspidites Black-headed VI X Taurine Mostly pythoca melanocephalus python Aspidites Woma python VI X Taurine PythoCA, CA Shows pythoca to CA transition ramsayi Liasis albertsii D Albert s VI X Taurine Mostly pythoca python Morelia boeleni Boehlin s VI X Taurine Mostly pythoca python Morelia spilota Carpet python VI X Taurine Mostly pythoca variegata Morelia spilata Diamond VI X Taurine Mostly pythoca python Morelia viridis Green tree VI X Taurine Mostly pythoca python Python bivittatus Burmese VI X Taurine Mostly pythoca python Python curtus Blood python VI X Taurine Mostly pythoca Python molurus Indian rock VI X Taurine Mostly pythoca molurus python Python regius Ball python VI X Taurine Mostly pythoca Pytho sebae African rock python VI X Taurine Mostly pythoca Uropeltidae Rhinophis sp. Shield-tail VI X Taurine PythoCA, CA Viperidae
Additional file 1 (cont.), p. 14 Conjugation Major bile s Methodology Comments Agkistrodon contortix contortix Agkistrodon contortix pictigaster Azemiops feae Copperhead VI X Taurine CA Trans-Peco copperhead VI X Taurine CA White-headed VI X Taurine CA viper Bitis arietans Puff adder VI X Taurine CA, CDCA Bitis atropos Mountain adder VI X Taurine Mixture of and C 23 bile, some with Bitis cornuta Eastern homsman adder VI X Taurine Mixture of and C 23 bile, some with Bitis gabonica Gaboon viper VI X Taurine Mixture of and C 23 bile, some with Bitis nasicornis Bothriechis schlegelii Bothrops alternates Bothrops jaracara Bothrops neuwiedi Cerastes cerastes Crotalus adamanteus Crotalus catalinensis Crotalus cerastes Rhinoceros viper VI X Taurine Mixture of and C 23 bile, some with Eyelash viper VI X Taurine CA Urutu VI X Taurine CA Jaracara VI X Taurine CA Neuwied s lancehead Homed sand viper VI X Taurine CA VI X Taurine Mixture of and C 23 bile, some with VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated Eastern diamondback Santa Cantalina and Δ22 bile Sidewinder VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile
Additional file 1 (cont.), p. 15 Conjugation Major bile s Methodology Comments Crotalus durissus Crotalus helleri Crotalus lepidus klauberi Crotalus ruber South American rattle South Pacific rattle Banded rock rattle Red diamond rattle Mojave green rattle Arubu Island rattle Uracoan rattle VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile Crotalus scutulatus Crotalus unicolor Crotalus vegrandis Daboia russelli Russell s viper VI X Taurine Mixture of and C 23 bile, some with Deinagkistrodon acutus Eristicophis macmahoni Snub-nosed pit viper Leaf-nosed viper VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile VI X Taurine Mixture of and C 23 bile, some with Lachesis muta Bushmaster VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile Macrovipera lebetina Turan bluntnosed viper VI X Taurine Mixture of and C 23 bile, some with Parias flavomaculatus Parias mcgregori Trimeresurus trigonocephalus Tropidolaemus wagleri CROCODYLIA Philippine VI X Taurine Mostly CA Small amounts of 23R-hydroxylated viper and Δ22 bile McGregor s VI X Taurine Mostly CA Small amounts of 23R-hydroxylated tree viper and Δ22 bile Sri Lanka viper VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile Wagler s viper VI X Taurine Mostly CA Small amounts of 23R-hydroxylated and Δ22 bile
Additional file 1 (cont.), p. 16 Conjugation Major bile s Methodology Comments Alligatoridae Alligator mississipiensis Alligator sinensis Caiman crocodilus Caiman latirostris Melanosuchus niger Paleosuchus palpebrosus Crocodylidae Crocodylus acutus Crocodylus intermedius Crocodylus johnsoni Crocodylus niloticus Crocodylus novaeguineae Crocodylus palustris American alligator Chinese alligator Spectacled caiman Broad-nosed caiman IV X Taurine Mostly 3α,7α,12αtrihydroxy-- cholestan-27-oic Black caiman Dwarf caiman American crocodile Orinoco crocodile Johnson s crocodile Nile crocodile New Guinea crocodile Mugger IV X Taurine Mostly 3α,7α,12α- trihydroxy--
Additional file 1 (cont.), p. 17 Conjugation Major bile s Methodology Comments Crocodylus porosus Crocodylus siamensis Osteolaemus tetraspis Australian water crocodile Siamese crocodile Dwarf crocodile cholestan-27-oic Gavialidae Gavialis gangeticus Tomistoma schlegelii Indian gavial False gavial