doi: http://folia.paru.cas.cz Research Article Cryptosporidium testudinis sp. n., Cryptosporidium ducismarci Traversa, 2010 and Cryptosporidium tortoise genotype III (Apicomplexa: Cryptosporidiidae) in tortoises,,,,, 4, Lada, and 4 Cryptosporidium number of studies on these hosts. The aim of the present study was to characterise the genetic diversity and biology of cryptosporidia in screened for presence of cryptosporidia by microscopy using aniline-carbol-methyl violet staining, and Cryptosporidium - Cryptosporidium C. ducismarci Cryptosporidium C. ducismarci are genetically distinct from previously described species of Cryptosporidium. Oocysts of Cryptosporidium are morphologically distinguishable from C. ducismarci Cryptosporidium C. ducismarci Gray) were infectious Mauremys reevesii [Gray]), common garter snake ( [Linnaeus]), Taeniopygia guttata Cryptosporidium C. ducismarci and biological data support the establishment of Cryptosporidium Cryptosporidium testudinis sp. C. ducismarci as a separate species of the genus Cryptosporidium. The genus Cryptosporidium species of protist parasites that infect epithelial cells in the microvillus border of the gastrointestinal tract of all classes Cryptosporidium have been under intensive investigation for more species infecting humans, livestock and other mammals, with comparatively little attention paid to cryptosporidia in cies of Cryptosporidium have been described best in snakes of Cryptosporidium in tortoises remains poor. the microscopic detection of oocysts in the faeces of an croscopy as detection methods, oocysts of cryptosporidia - Zoobank number for article: This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Cryptosporidium in tortoises Table 1. Occurrence of species of Cryptosporidium Cryptosporidium toise Cryptosporidium in other reptiles and environmental samples. Groups Cryptosporidium spp. Country Tortoises Turtles Other reptiles (radiated tortoise) (gopher tortoise) association number) Cryptosporidium sp. e C. testudinis sp. n. Cryptosporidium sp. Cryptosporidium sp. Cryptosporidium sp. Cryptosporidium sp. Cryptosporidium sp. Indotestudo sp. Cryptosporidium sp. Testudo hermanni Gmelin Gray Lortet Testudo marginata (marginated tortoise) (Linnaeus) (green turtle) (bog turtle) Python regius (ball python) (veiled chameleon) C. ducismarci Traversa, C. testudinis sp. n. C. ducismarci Cryptosporidium sp. C. testudinis sp. n. C. ducismarci Cryptosporidium sp. C. ducismarci C. parvum Cryptosporidium sp. Cryptosporidium sp. C. testudinis sp. n. C. ducismarci C. ducismarci C. testudinis sp. n. were detected in faecal samples of various tortoise species to describe the Cryptosporidium tortoise genotype (later called Cryptosporidium- of C. parvum Cryptosporidium Cryptosporidium Cryptosporidium the name Cryptosporidium ducismarci Cryptosporidium Cryptosporidium species a result, many authors do not consider C. ducismarci to be Cryptosporidium infection in tortoises to date is provid- transmission, oocyst morphology and molecular characteristics of Cryptosporidium C. ducismarci studies, which show that Cryptosporidium tortoise geno- Cryptosporidium also provide previously unreported data on C. ducismarci, which is recognised as a valid species. Tortoise species owned by private breeders, pet shops and
Cryptosporidium in tortoises Cryptosporidium Tyzzer, Cryptosporidium testudinis sp. n., Cryptosporidium ducismarci Cryptosporidium tortoise genotype - The faecal consistency (loose if it took the form of the container and solid if it maintained its original shape) was noted at animals were screened without previous knowledge of parasitological status. Oocysts of Cryptosporidium C. ducismarci were originally isolated from faecal samples of naturally Gray). The tortoise infected with Cryptosporidium infected with C. ducismarci oocysts of cryptosporidia following aniline-carbol-methyl violet - - - Oocysts of Cryptosporidium C. ducismarci - Cryptosporidium - - ed. As a control, the morphometry of C. parvum Bos taurus Linnaeus) was measured by the same person using the same microscope. Cryptosporidium
Cryptosporidium in tortoises C. parvum was used as - - each locus were repeated two times. - - - - CryptosporidiumCryptosporidium testudinis sp. n., Cryptosporidium ducismarci Cryptosporidium scale bar indicate number of substitution per site. C. ducismarci - - ( Cryptosporidium oocyst wall protein - -
Cryptosporidium in tortoises Cryptosporidium Cryptosporidium spp., including Cryptosporidium testudinis sp. n. and Cryptosporidium ducismarci of substitution per site. - - Mauremys reevesii Mus mus- Linnaeus), an adult common garter snake (Thamnophis Taeniopygia guttata with Cryptosporidium C. ducismarci. Three sporidia. Cryptosporidium-positive animals were additionally sampled weekly - spectively. Consistency and colour of faeces and intensity of the Ten samples were positive by microscopy, with an infection Cryptosporidium - - clusters among isolates of cryptosporidia from tortois- the clusters included Cryptosporidium, cies in the present study. A second cluster included C. ducismarci, previously reported from a marginated tortoise (Testudo marginata A third cluster included a single isolate from a Leopard tortoise ( in the present study and an isolate from was most closely related to Cryptosporidium tortoise geno- - Cryptosporidium
Cryptosporidium in tortoises Cryptosporidium Cryptosporidium oocyst wall protein genes. Tortoise n Cryptosporidium spp. Actin C. testudinis sp. n. C. ducismarci (Miller) (sulcata tortoise) - 0/0 - - - - - 0/0 - - - - (Gray) (Chaco tortoise) C. testudinis sp. n. - 0/0 - - - - - 0/0 - - - - - 0/0 - - - - C. ducismarci (Kuhl) (serrated tortoise) 4 C. testudinis sp. n. C. testudinis sp. n. C. ducismarci - Testudo graeca Linnaeus (Greek tortoise) C. testudinis sp. n. C. ducismarci Testudo hermanni C. testudinis sp. n. C. ducismarci 4 Testudo C. testudinis sp. n. C. ducismarci 4 Testudo C. ducismarci Testudo marginata C. testudinis sp. n. C. ducismarci 4 Terrapene - 0/0 - - - - Total C. testudinis sp. n. - C. ducismarci - 0 - - Cryptosporidium tortoise Two morphotypes of oocysts were detected in screened faecal samples. On the basis of morphometrics, oocysts of Cryptosporidium larger than oocysts of C. ducismarci Cryptosporidium data on C. ducismarci Cryptosporidium testudinis Oocysts are shed fully sporulated with ± developmental stages unknown. Type host: ( Gray). breeder). Location in the host unknown. O t h e r h o s t s : chaco tortoise ( [Gray]), Greek tortoise (Testudo graeca (Testudo hermanni ), leopard tortoise ( ), marginated tortoise (Testudo marginata), radiated tortoise ( radiata [Kuhl]). - testudinis is derived from the Oocysts are larger than those of C. ducismarci reagents developed primarily for C. parvum ferentiated genetically from other cryptosporidia based on
*6 *==**M >6*J 6'7 Cryptosporidium testudinis & Cryptosporidium in tortoises Cryptosporidium ducismarci 0 /2 92 Ooocysts of Cryptosporidium testudinis sp. n. and Cryptosporidium ducismarci V >6*6 # 8 es ( Gray). Oocysts visualised in various preparations. A \ ~ # D B \ 9 H 9 H#! D C \ 9 HCryptosporidium E V H 3 9 0 /2 :2 Course of infection of Cryptosporidium testudinis sp. n. and Cryptosporidium ducismarci V >6*6 8 (? 9 # F # X KU black circle indicates microscopic detection of oocysts. * 2 GF # W 9! a 8 ; ), but not 8 ` tles (Mauremys reevesii), a common garter snake (Tham? " 9 X! ;Taeniopygia guttata) or $ K # ; ). $ X KU C. testudinis W X ** K #! W # 76 K ;E 7? W & # >66 K W!! was terminated (data not shown). Oocysts of C. testudinis W 9 #! F # W!! F # 9 '7 K W!!! * 666 Y F # #! F! 9 W!! W! " W! U! W U! # W! Cryptosporidium ducismarci V >6*6 E = 7 * ( - 2 Oocysts are shed fully sporulated (four sporozoites and oocyst residuum inside) and measure E >6*J J' 6'7 4 =\7 = ]# ;# [ 7 6 ]#? ^ = '\7 ' ]# ;# [ = @ ]#? W!!MW! * * ± 6 6' ; [ '6? 8 ( / 2 Oocysts of C. ducismarci are smaller than those of C. testudinis and indistinguishable from those of C. parvum! # S to other species of Cryptosporidium and cross react with ## # C. parvum. ( - (& $ W! KU!! "! # $ "! 8 9 Q $$N YP W & ; U }'=76*@\ }'=76>J }'=76'5\ }'=76=7 }'=7677\ }'=76J'? * 2 >66@ Cryptosporidium genotype named Cryptosporidium W! ~ ;V >66@ >6*6?! X! Cryptosporidium! $$N YP Q!! 5 *6
Cryptosporidium in tortoises Cryptosporidium ducismarci lacked description of oocyst morphology. Therefore, it was not be considered as a valid species by some authors. This article redescribes C. ducismarci by providing additional morphological, biological and molecular data to support its established in a ), but not Mauremys reevesii), a common garter snake ( Taeniopygia guttata ). C. ducismarci - not shown). Cryptosporidium testudinis, C. ducismarci and Cryptosporidium ly, which is comparable to data provided by Traversa et C. testudinis, i.e. C. ducismarci were performed on captive tortoises and the occurrence in wild animals is not known. The morphology of oocysts of C. testudinis and C. ducismarci is typical of those of species of Cryptosporidium C. testudinis C. ducismarci, which makes it possible to distinguish these species microscopi- Cryptosporidium, including C. serpentis, which are oval C. testudinis are spherical. Other characteristics of oocysts of C. testudinis and C. ducismarci, including thickness of the wall, its inner structure and ability to be detected using Cryptosporidium guish C. testudinis and C. ducismarci from other species of Cryptosporidium C. testudinis (reported as Cryptosporidium Python regius tus Cryptosporidium, that is described herein as C. testudinis and C. ducismarci demonstrated that an uncharacterised Cryptosporidium inoculum, prepared from the combined faeces of a naturally - was infectious for black rat snakes, C. testudinis and C. ducismarci produced no clinical signs in tortoises, which contrasts with previous reports of symptoms such as weight loss, weakness, lethargy, pneumonia, apathy, depression, innapetence, dehydration, diarrhoea and edema of the head and neck in tortoises infected with C. testudinis (referred as Cryptosporidium C. ducismarci (referred as Cryptosporidium - clinical signs could have been due to the presence of other of clinical signs during infection by C. testudinis study, only tortoises infected with C. parvum (referred as C. pestis of eight Cryptosporidium were also positive for, Proteus sp. (both Hexamita - tortoise and pancake tortoise infected with C. ducismarci and other pathogens such spp. showed mod- cytes in Cryptosporidium infection in - C. testudinis and C. ducismarci in C. testudinis were C. ducismarci was never detected in the faeces by this approach. This is probably due to low number of oocysts being shed and the - - - C. serpentis Cryptosporidium spp., such as C. tyzzeri - C. erinacei hedgehogs, and C. ryanae caused by C. testudinis and C. ducismarci are characterised by low oocyst shedding for a prolonged period with- C. varanii and C. serpentis - -
Cryptosporidium in tortoises C. testudinis and C. ducismarci are genetically distinct C. testudinis and C. ducismarci from C. varanii, respectively. At the actin locus, C. testudinis and C. ducismarci C. serpentis and C. varanii, respec- C. testudinis and C. ducismarci C. muris and ences are much greater than those between closely related Cryptosporidium C. parvum and C. tyzzeri spectively, and distances between C. muris and C. andersoni - for providing data and samples for our research. characterization of Cryptosporidium spp. in mammals and rep- methods for calf-derived Cryptosporidium parvum oocysts us- karyot. Microbiol. Cryptosporidium in snakes with hypertrophic elaphid snakes with Cryptosporidium serpentis Cryptosporidium Cryptosporidium ryanae Bos taurus). n. Bos taurus). J. Cryptosporidium sp. infections in green turtles,, as a potential source viron. Microbiol. sion of Cryptosporidium oocyst isolates from mammals, birds Cryptosporidium tortoise, - - - - Cryptosporidium genotypes in storm event water samples from three - Cryptosporidium serpentis in a wild-caught corn snake ( ) and assay that can additionally discern C. parvum from C. wrairi. tosporidiidae): molecular and biological evidence of cryptic species within gastric Cryptosporidium Cryptosporidium erinacei - Cryptosporidium pigs ( - Cryptosporidium pher tortoise ( ) from eight populations in sporidia by aniline-carbol-methyl violet and tartrazine in smears logical analysis of snakes naturally infected with Cryptosporidium serpentis ducing reptiles into a captive collection: the role of the veterinar-
Cryptosporidium in tortoises - of Cryptosporidium ceedings of the Annual meeting of the American Association of Cryptosporidium tyzzeri mice ( Testudo hermanni Cryptosporidium gen- - Cryptosporidium - Cryptosporidium Giardia and evolutionary relationships of Cryptosporidium parasites at - tis in leopard geckos ( ) associated with Cryptosporidium Cryptosporidium infecting tortoises: Cryptosporidium ducismarci tors Cryptosporidium from tortoises: Genetic characterisa- Cryptosporidium Genetic diversity within Cryptosporidium parvum and related Cryptosporidium of Cryptosporidium crobiol. adaptation and host-parasite co-evolution in Cryptosporidium: Cryptosporidium - Cryptosporidium testudinis sp. n., Cryptosporidium ducismarci Cryptosporidium