Species and Genotypes of Cryptosporidium RON FAYER USDA AGRICULTURAL RESEARCH SERVICE WHAT IS A ?

It is a basic unit of biological classification and a taxonomic rank. A species is often defined as a group of organisms capable of interbreeding and producing fertile offspring. More precise or differing measures can be used, such as similarity of DNA, morphology or .

Species that are believed to have the same ancestors are grouped together, as a .

All species have two part name (a "binomial name"). The first part of a binomial name is the genus of the species. The second part is the specific name.

For example, Boa constrictor which is commonly called by its binomial name, and is one of four species of the Boa genus.

The first part of the name is capitalized, and the second part has a lower case. The two part name is written in italics. What are the sources of Cryptosporidium?

FIELD MOUSE

WOODCHUCK

VOLE

SKUNK

BEAVER

More than 150 species of infected with Cryptosporidium spp. Artiodactyla nasomaculatus (Addax) Aepyceros melampus () elaphus (Red //wapiti) Ammotragus lervia (Barbary ) Cervus eldi (Eld's deer) Alces alces () Cervus nippon () Antidorcas marsupialis () Cervus unicolor (Sambar) Antilocapra americana () Connochaetes gnou () Antilope cervicapra () Connochaetes taurinus (Blue-eared gnu) Axis axis (Axis deer) dama (Fallow deer) bison () Elaphurus davidianus (Pere David’s deer) Bison bonasus () Gazella dama (Addra ) indicus () Gazella dorcas (Dorca's gazelle) Bos taurus () Gazella leptoceros (Slender-horned gazelle) Boselaphus tragocamelus () Gazella subgutterosa (Persian gazelle) bubalis (Water ) Gazella thomsoni (Thomson's gazelle) Bubalus depressicornis (Lowland ) Giraffa camelopardalis () Camelus bactrianus (Bactrian ) Hexaprotodom liberiensis (Pygmy hippopatomus) falconeri (Turkomen ) niger (Sable ) Capra hircus () ellipsiprymmus (Ellipsen ) capreolus () glama () Cervus albirostris (Thorold's deer) Lama guanicoe () Cervus duvauceli ( deer)

Muntiacus reevesi ( deer) hemionus ( deer) Lontra canadensis (river otter) Odocoileus virginianus (White-tailed deer) Panthera pardus () gazella callotys (Fringe-eared oryx) Procyon lotor () Oryx gazella dammah (Scimitar horned oryx) Urocyon cinereoargenteus (Grey fox) aries (Sheep) Ursus americanus (Black bear) Ovis musimon () Ursus arctos () Ovis orientalis () Ursus (Thalarctos) maritimus () Sus scrofa () Vulpes vulpes () Syncerus caffer () Zalophus californianus (California sea ) oryx (Eland) Tayassu tajacu (Collared ) Order Chiroptera eurycerus () Eptesicus fuscus (Big brown bat) Order Carnivora Myotis adversus (Large-footed mouse-eared bat) Acironyx jubatus () Canis familiaris () Order Insectivora Canis latrans () Ateletrix albiventris (African ) russula (Greater white-toothed ) Felis catus () Erinaceus europaeus () MartesHelarctos foina malayanus (Beech marten) (Malayan bear ) Meles meles (Badger) araneus (Long-tailed shrew) Mephitis mephitis (Striped skunk) Sorex minutus (Pygmy shrew) Mustela putorius () Order Mustela vison (American mink) Oryctolagus cuniculus () Sylvilagus floridanus (Cottontail)

Order Marsupialia Cercocebus torquatus (White-collared monkey) Antechinus stuartii (Brown antechinus) Cercopithecus aethiops (Velvet monkey) Didelphis virginiana () Erythrocebus patas (Patas monkey) Isodon obesulus (Southern brown bandicoot) Eulemur macaco (Black lemur) Macropus giganteus (Eastern grey kangaroo) Gorilla gorilla (Gorilla) Macropus rufogriseus (Red neck wallaby) Homo sapiens (Humans) Macropus rufus (Red kangaroo) Hylobates syndactylus syndactylus (Siamang) Phascolarctos cinereus (Koala) Lemur catta (Ring-tailed lemur) Thylogale billardierii (Pademelon) Lemur macacomayottensis (Brown lemur) Trichosurus vulpecula (Brushtail possum) Lemur variegatus (Ruffed lemur) Macaca fascicularis (Long-tailed macaque) Order Perissodactyla Macaca fuscata (Japanese macaque) Ceratotherium simum (Southern white rhinoceros) Macaca mulatta (Rhesus monkey) Equus caballus () Macaca nemestrina (pigtail macaque) Equus przewalski (Miniature horse) Macaca radiata (Bonnet macaque) Equus (Zebra) Macaca thibetana (Pere David s macaque) Rhinoceros unicornis (Rhinoceros) Mandrillus leucophaeus (Drill) Tapirus terrestris (Brazilian ) Nycticebus pygmaeus (Lesser slow loris) Papio anubis (Olive ) Order Papio cynocephalus (Baboon) Ateles belzebuth (Marimonda spider monkey) Pithecia pithecia (White-faced saki) Calithrix jacchus (Common marmoset) Pongo pygmaeus (Orangutan Cercopithecus campbelli (Campbell's mona) Saguinus oedipus (-topped tamarin) Cercopithecus talapoin (Talapoin monkey) Saimiri sciureus (Squirrel monkey) Varecia variegata (Red-ruffed lemur) Cercocebus albigena (Mangabey) Sciurus niger () Sigmodon hispidus (Cotton rat) Order Rodentia Spermophilus beecheyi (California ground Apodemus agrarius (Field mouse) squirrel) Apodemus flavicollis (Field mouse) Spermophilus tridecemlineatus Apodemus sylvaticus (Field mouse) (13-lined ground squirrel) Castor canadensis (Beaver) Mus spretus (Western Mediterranean mouse) Castor (European beaver) Tamias sibiricus (Siberian chipmunk) porcellus ( pig) Tamias striatus (Chipmunk) laniger (Chinchilla) Clethrionomys glareolus (Red-backed vole) Order Monotremata Coendou prehensilis (ring tailed porcupine) Tacyglossus aculeatus (Echidna) Geomys bursarius (Pocket gopher) Glaucomys volans (Flying squirrel) Order Proboscidea Hystrix indica (Indian porcupine) Elephas maximus (Indian elephant) Marmota monax (Woodchuck) Loxodonta africana () Mesocricetus auratus (Golden ) Microtus agrestis (Field vole) Order Sirenia Microtus arvalis (Orkney vole) Dugong dugon (Dugong) Mus musculus (House mouse) Myocastor coypus (Coypu) Order Pinnipedia Ondatra zibethicus (Muskrat) Mirounga leonina (Southern elephant seal) Rattus norvegicus (Norwegian rat) Phoca hispida (Ringed seal) Rattus rattus (House rat) Zalophus californianus (California sea lion) Sciurus carolinensis (Gray squirrel)

MICROSCOPIC BASIS FOR DETECTION OF CRYPTOSPORIDIUM SELECTED SPECIES OF CRYPTOSPORIDIUM

Modified from Xiao et al. (2004) Length (μm) Width (μm) L:W Ratio No. of oocysts Parasite measured Mean Mean Mean C. parvum 44 4.85 4.39 1.11 C. hominis 44 4.91 4.28 1.15

C. meleagridis 55 4.93 4.40 1.12

Pig genotype I 56 5.05 4.41 1.15

C. saurophilum 20 4.94 4.49 1.14

Opossum 55 5.26 4.38 1.20 genotype I

Greatest Size Differences 0.41μm 0.21μm 0.09

Modified from Xiao et al. (2004) SPECIES OF CRYPTOSPORIDIUM (based on oocyst morphology, molecular, and host specificity data)

REPTILES/ FISH AMPHIBIANS BIRDS

C. scophthalmi C. serpentis C. meleagridis C. hominis C. molnari C. varanii C. baileyi C. parvum C. ducimari C. galli C. ubiquitum C. fragile C. felis C. canis C. muris C. suis C. cuniculus C. andersoni C. fayeri C. macropodum C. xiaoi C. ryanae C. bovis C. wrairi C. tyzzeri SPECIES OF CRYPTOSPORIDIUM (based on oocyst morphology, molecular, and host specificity data)

REPTILES/ FISH AMPHIBIANS BIRDS MAMMALS

C. scophthalmi C. serpentis C. meleagridis C. hominis C. molnari C. varanii C. baileyi C. parvum C. ducimari C. galli C. ubiquitum C. fragile C. felis C. canis C. muris C. suis C. cuniculus C. andersoni C. fayeri C. macropodum C. xiaoi C. ryanae C. bovis C. wrairi C. tyzzeri Genotypes of Cryptosporidium

39 selected genotypes C. sp. Bear C. sp. Pig II C. sp. Deer C. sp. Raccoon C. sp. Deer mice C. sp. Rat I, II, III C. sp. Duck C. sp. Seal 1 and 2 C. sp. Ferret C. sp. Sheep novel genotype C. sp. Goat (Chalmers et al and Ryan et al) C. sp. Fox and Fox II C. sp. Chipmunk I, II,III C. sp. Goose I and II C. sp. Shrew C. sp. Hamster C. sp. Skunk C. sp. Horse C. sp. C. sp. Mongoose C. sp. Caribou (gastric) C. sp. Monkey C. sp. Lizard (gastric) C. sp. Muskrat I and II C. sp. Tortoise (gastric) C. sp. Opossum I and II C. sp. (gastric) C. sp. Ostrich C. muris Japanese field mouse

Most human are caused by C. hominis and C. parvum

12 other species or genotypes also detected in immunocompetent and/or immunocomprised humans:

C. meleagridis C. felis C. canis C. suis C. muris C. andersoni C. ubiquitum C. cuniculus C. fayeri and the horse, skunk, and monkey genotypes

(Pieniazek et al., 1999; McLauchlin et al., 2000; Ong et al., 2002; et al., 2003; Mallon, et al., 2003; Learmonth et al., 2004; Leoni et al., 2006; Chalmers et al., 2009; Robinson et al., 2010). PROCESS OF NAMING A SPECIES OF CRYPTOSPORIDIUM

1- Morphometric studies of oocysts 2- Genetic characterizations 3- Demonstration of natural and, when possible, at least some experimental host specificity 4- Compliance with ICZN PROCESS OF NAMING A SPECIES OF CRYPTOSPORIDIUM

1- Morphometric studies of oocysts 2- Genetic characterizations 3- Demonstration of natural and, when possible, at least some experimental host specificity 4- Compliance with ICZN GENETIC CHARACTERISTICS

8 10.4 Mb

SSUrRNA (Small sub unit ribosomal RNA) COWP (Crypto oocyst wall ) ACTIN HSP (heat shock ) TRAP C-1, C-2 (thrombospondin related adhesive proteins) b-TUBULIN GP60 (Glyco protein) GP40/15 ACETYL COA DHFR (dihydrate folate reductase) GENETIC CHARACTERISTICS

8 CHROMOSOMES 10.4 Mb

SSUrRNA (Small sub unit ribosomal RNA) COWP (Crypto oocyst wall protein) ACTIN HSP (heat shock proteins) TRAP C-1, C-2 (thrombospondin related adhesive proteins) b-TUBULIN GP60 (Glyco protein) GP40/15 ACETYL COA DHFR (dihydrate folate reductase) GENETIC CHARACTERISTICS

8 CHROMOSOMES 10.4 Mb

SSUrRNA (Small sub unit ribosomal RNA) COWP (Crypto oocyst wall protein) ACTIN HSP (heat shock proteins) TRAP C-1, C-2 (thrombospondin related adhesive proteins) b-TUBULIN GP60 (Glyco protein) GP40/15 ACETYL COA DHFR (dihydrate folate reductase) SUBTYPING

Based on sequence analysis of the 60-kDa glycoprotein (gp60)

Used in tracking the transmission of six Cryptosporidium species and genotypes, including C. hominis, C. parvum, C. meleagridis, C. fayeri, and the rabbit and horse genotypes

There are at least 10 gp60 subtype families of C. parvum, two (IIa and IId) of which are involved in zoonotic transmission TIME APRIL 19,1993 Cryptosporidium hominis

Humans * Sheep** ** Dugong

*natural and experimental ** experimental Cryptosporidium parvum

Humans Domesticated animals: Cattle Sheep Mice Rats Wild animals: White tail deer Mice Wildebeest Cryptosporidium meleagridis

HOSTS

Turkey Human* MOST OFTEN Chicken Parrots Cockatiels Pigs Rats Rabbits Mice Cattle

* 3rd most prevalent spp. in humans * IMMUNOCOMPETENT 36% * IMMUNOCOMPROMISED 64% Cryptosporidium felis

HOSTS: CAT HUMAN* 97 cases COW *Lucio-Forster et al. 2010 Trends in Parasitology 26:174

Estimated 70 million and homeless stray in the US The Humane Society Jan 2011 Cryptosporidium canis HOSTS: FOXES CATTLE HUMANS* 28 cases

*Lucio-Forster et al. 2010 Trends in Parasitology 26:174

~75 million dogs in US homes, no. of feral dogs unknown Cryptosporidium muris

MICE RATS RABBITS GERBILS GUINEA PIGS DOGS CAMEL CALVES HUMANS (mmunocompromised) Cryptosporidium ubiquitum (formerly: cervine genotype)

HUMANS* WHITE TAIL DEER SHEEP CATTLE SIKA DEER IBEX BOER GOAT MOUFLON BEAVER MICE GERBILS CHIPMUNK WOODCHUCK RACCOON DEER MOUSE SQUIRREL COENDU LEMURS * 9 immunocompetent > 500 Wild Samples from Chesapeake Watershed

Species Crypto Zoonotic

Beaver 1% Muskrat 11% Otter 0% Fox 8% C. canis (5) Raccoon 4% C. sp. skunk (2) White-tailed Deer 58%

Atlantic CoastSHELLFISH Shellfish Survey 13 States IFA PCR 38 Sites Maine + + Massachusetts + + Connecticut + + Rhode Island + + + - New Jersey + + Delaware - - Maryland + + Virginia + + + - South Carolina + - - - + + Collection No. Positive Province Species/genotype Site Location IFA PCR or State or New NW Gulf of St. 1 O 3/25 1/5 C. parvum Brunswick Lawrence 3 Maine O Blue Hill Bay 1/25 2/5 C. hominis Piscatagua River C. parvum 4 Maine O 1/25 1/5 (Conn.) C. parvum CRYPTO 5 Mass. O Cape Cod Bay 1/25 5/5 C. meleagridis IN U.S. 6 Mass. O South Cape Cod 2/25 5/5 C. parvum C. parvum COASTAL 7 Mass. C South Cape Cod 0/25 5/5 C. meleagridis C. parvum WATERS 8 Mass. C South Cape Cod 0/25 5/5

Rhode Upper Narragansett 9 O 1/25 2/5 C. parvum Island Bay

Long Island Sound, C. hominis 13 Connecticut O 0/25 1/5 Norwalk C. meleagridis

Long Island Sound, C. hominis 14 Connecticut C 0/25 1/5 Milford C. meleagridis C. parvum 21 New Jersey C Great Egg Harbor 1/25 1/5

25 Maryland O Big Choptank River 0/25 1/5 C. parvum

26 Virginia O James River 0/25 1/5 C. parvum Inter Coastal Fayer et al 2003. 37 Fla. O Waterway, St. 0/25 1/5 C. parvum Parasitol Res 89:141. Augustine SUMMARY

DEFINITION OF SPECIES

SOURCES OF WATERBORNE CRYPTOSPORIDIUM

150 HOSTS

NAMES OF CRYPTOSPORIDIUM SPECIES AND GENOTYPES

THE NAMING PROCESS

CRYPTO USED FOR

7 SELECTED SPECIES AND THEIR HOST RANGES

WILDLIFE IN WETLAND AREAS DRAINING INTO CHESAPEAKE BAY

ATLANTIC COAST SHELLFISH CONTAMINATED WITH CRYPTOSPORIDIUM