Testudo Graeca Research Articles

Carapace Morphology Variations in Captive Tortoises: Insights from Three-Dimensional Analysis.

The carapace morphology of tortoises is a crucial characteristic used for species identification, with features such as shell shape, roughness, and color patterns varying among species. Understanding this morphological diversity is valuable not only for taxonomic classification but also for more specialized clinical approaches. This study investigated the morphological differences in the shells of Leopard tortoises (Stigmochelys pardalis), African spurred tortoises (Centrochelys sulcata), and Greek tortoises (spur-thighed tortoises; Testudo graeca) raised in captivity. Using 3D scanners, the carapaces were modeled, and a 3D geometric morphometric method was employed to analyze shape variations and dimensional features, with landmarks applied automatically. Among the species studied, African spurred tortoises had the largest carapace size. Principal component analysis (PCA) identified PC1 and PC3 as critical factors in distinguishing between species based on morphological characteristics. Positive PC1 values, associated with a shorter carapace height, indicated a flatter or more compact shell shape. A higher PC3 value corresponded to a raised shape at the back of the shell, while a lower PC3 value indicated a raised shape at the front. Specifically, Leopard tortoises exhibited a higher carapace shape than the other species, while African spurred tortoises had shorter carapaces. An allometric effect was observed in the carapaces, where smaller specimens tended to be proportionately higher-domed, whereas larger shells displayed a lower height in shape. These findings highlight the significance of shape variations in tortoise shells, which emerge during adaptation and have important implications for taxonomy and clinical practice. Such differences should be carefully considered in veterinary care and species identification.

Tick Infestation of the Mediterranean Spur-Thighed Tortoises (Testudo graeca, Linnaeus, 1758) from Western Regions of Algeria

Abstract The Spur-thighed tortoise has a large geographic distribution and can be infested by several tick species, mainly Hyalomma aegyptium. This tick has recently been confirmed as a natural carrier of numerous pathogenic, including zoonotic agents. The purpose of this study was to estimate the level of hard tick infestation in Mediterranean tortoises living in some regions of the west of Algeria (Oran, Mostaganem, and Tiaret) during the spring and summer of the year 2019. Among 39 turtles collected from the different study regions, 22 tested positive, which represents an infestation rate of 56.41 %. A total of 224 adult ticks and one nymph were collected, which can display a tick infestation intensity of 10.23 %. All of these ticks belong to a single species, which is Hyalomma aegyptium. In terms of abundance, this study showed 5.78 ticks/tortoise. Regarding sex, we have collected 198 male and 26 female ticks with a sex ratio of 7.62. The preferred attachment sites of Hyalomma aegyptium ticks were the inguinal, the limbs, and to a lesser degree, the neck. A few rare cases have been collected from the carapace. Further studies are needed to assess the impact of the intensity of H. aegyptium infestation on the tortoise’s health status and to assess the potential transmission of zoonotic pathogens.

Macro- and micro-anatomical investigation of the oropharyngeal roof of landform greek tortoise (Testudo graeca graeca) and semi-aquatic red-eared slider turtle (Trachemys scripta elegans)

The present investigation examined the oropharyngeal roof of two turtles having different feeding behaviors: the landform Greek tortoise (Testudo graeca graeca) primarily herbivores and the semi-aquatic red-eared slider turtle (Trachemys scripta elegans) lives in freshwater that opportunistic omnivorous grossly and by scanning and light microscopes. Grossly, the Greek tortoise had a V-shaped roof consisting of the upper rhamphotheca, peri-palatine region, upper alveolar ridge, peripheral palatine ridge, median palatine ridge, vomer, choanae, caudal palatine part, and pharynx. At the same time, the red-eared slider had a semilunar roof consisting of upper rhamphotheca, two peripheral palatine ridges, core of palatine ridges, upper alveolar band, vomer, choanae, caudal palatine part, and pharynx. SEM revealed that the red-eared slider roof appeared more straightforward. The upper rhamphotheca is sharp, with a median premaxillary notch in the red-eared slider that gives a powerful bite for cutting to compensate absence of the teeth. Additionally, the red-eared slider’s upper alveolar band is interrupted by a single upper alveolar ridge that appears spiky, pointed, and longer as it needs powerful chewing of prey and there are two types of teeth-like projections at its peri-palatine area for food-crushing and chewing. The Greek tortoise palatine region had numerous ridges and folds to provide roughness for food processing. Greek tortoises had small-sized choanae with two choanal folds to minimize choanal openings when eating dusty grasses. Histologically, Greek tortoise palate was rostrally thicker and more keratinized than caudally, and the caudal palatine region was characterized by a single pair of circumvallate-like papilla with multiple mucous openings and secretions, while red-eared slider palate was slightly keratinized at the peri-choanal region, and the rest of the palate was non-keratinized with few mucous openings. The current investigation found various structural oropharyngeal roof adaptations to feeding behavior in the omnivore red-eared slide compared to the herbivorous Greek turtle.

Age estimation in tortoises: an evaluation of methods for the spur-thighed tortoise (Testudo graeca)

Abstract Age and growth-related parameters provide a better understanding of life-history adaptations and trade-offs and can be studied in the wild through capture-mark-recapture (CMR) or sclerochronology (i.e., skeletochronology and counting growth marks on hard structures). In chelonians, skeletochronology is limited to carcasses, CMR is limited by time and funding constraints, but counting shell growth annuli (SGA count) is a cheaper and faster alternative. A long-term CMR study in Testudo graeca from south-eastern Romania allowed us to compare the age estimated by the two sclerochronology methods using carcasses, and calibrate the results with data obtained from recaptured tortoises. We found that the accuracy of SGA counts significantly decreased with time (i.e., years between captures and older individuals). Skeletochronology showed the best results with long bones (i.e., ulna, radius, femur), but overall underestimated the age when compared to SGA counts; we obtained comparable results from these two methods in tortoises up to 17 years old. The oldest age estimated by skeletochronology alone was 28 years, 30 years by SGA counts, and 40 years by SGA counts when calibrated by CMR. The growth models using age estimations from both skeletochronology and SGA counts were similar, highlighting their usefulness in monitoring growth trajectories. Our study showed that while age estimation in tortoises is challenging, a combination of methods can improve its accuracy.

Description of cases of axial bifurcation of the Mediterranean Spur-thighed tortoise – Testudo graeca Linnaeus, 1758 (Testudinidae, Reptilia) based on field research and herpetological collection of ZIN RAS

Description of cases of axial bifurcation of the Mediterranean Spur-thighed tortoise – Testudo graeca Linnaeus, 1758 (Testudinidae, Reptilia) based on field research and herpetological collection of ZIN RAS

Macroscopic, microscopic, and immunofluorescent characterization of the Greek tortoise (Testudo graeca graeca) oropharyngeal floor with concern to its feed adaptation as a herbivorous land reptile.

The current investigation focuses on gross anatomy, light, and scanning electron microscopy (SEM) of the Testudo graeca oropharyngeal floor, with particular reference to the immunofluorescence technique to examine its tongue. The T. graeca oropharyngeal floor showed many anatomical structures: the lower rhamphotheca, paralingual ridge, lower alveolar ridge, tongue, laryngeal mound, and glottis. The lower rhamphotheca appeared as a V-shaped jaw line with a highly serrated edge and a median tomium (beak). SEM observations of the lingual apex and the lingual body showed rectangular and conical filiform papillae with porous surfaces and taste pores. Meanwhile, the lingual root had two wings that carried papillae with different shapes: dagger-shaped, conical, bifurcated, and leaf-like papillae, and these papillae lacked taste pores. The laryngeal mound had openings for the laryngeal mucus gland and its secretions. Light microscopy findings showed mucous glands in the propria submucosa and near the mucosal surface of the lingual apex. The lingual root had lingual papillae and two hyaline cartilaginous skeletons between skeletal muscles, and the lingual papillae were elongated filiform, rectangular filiform papillae, and fungiform papillae. The lamina propria constituted the core of the lingual papillae and the mucous gland, they had a positive reaction with the periodic acid schiff (PAS) reagent. The apical surface of the fungiform papillae had taste pores. Under immunofluorescence, the vimentin was detected in taste bud cells, and synaptophysin reacted to the taste buds and nerve bundles. The current study of the Greek tortoise oropharyngeal floor investigated its herbivorous eating habits using its serrated lower rhamphotheca, a large tongue with differently shaped papillae, and numerous mucous glands. RESEARCH HIGHLIGHTS: The Greek tortoise (T. graeca graeca) oropharyngeal floor showed many anatomical structures: lower rhamphotheca, paralingual ridge, lower alveolar ridge, tongue, laryngeal mound, and glottis. SEM and light microscopy observations of the tongue revealed varied types and shapes of lingual papillae with a porous surface on the tongue apex (rectangular or conical filiform papillae), on the tongue body (filiform and fungiform papillae), and on the tongue root (dagger-shaped, conical, bifurcated, and leaf-like papillae). Light microscopy findings: the lamina propria constituted the core of the lingual papillae and had numerous mucous glands that had a slightly magenta-red color with PAS reagent. The apical surface of the fungiform papillae had taste pores. Vimentin and synaptophysin gave a reaction to the taste buds.

Alternative vegetation trajectories through passive habitat rewilding: opposite effects for animal conservation

ContextPassive habitat rewilding after rural abandonment can affect wildlife differently depending on the type of habitats that it generates.ObjectiveEvaluate and compare the effects of two alternative vegetation trajectories that occur through passive habitat rewilding in Mediterranean ecotone areas (crop-scrub and crop-pine forest transitions) on the long-term population dynamics of animal species.MethodsWe used the spur-thighed tortoise (Testudo graeca), a characteristic long-lived species of cultural landscapes, as study species. We applied a spatially explicit and individual-based model (STEPLAND) to simulate the movement and demographic processes in a long-term period, by comparing an “impact scenario” (i.e., historical land-use changes) to a “control scenario” (no land-use changes).ResultsThe two landscape scenarios resulted in different population trends. In the crop-scrub scenarios (control and impact), population densities increased similarly over time. However, the crop-pine forest scenario negatively affected population density throughout the simulation period, and showed a time-lag response of three decades. The extinction risk was 55% with a time-lag response of approximately 110 years.ConclusionsOur study highlights the need to analyse the legacy effects on long-lived ectotherms, using them as a proxy to understand the future effects of dynamic landscapes created by “passive habitat rewilding”. Our results showed how traditional agriculture in Mediterranean ecotone areas may generate “ecotone effects” (i.e. increase in demographical parameters), but also population extinction on long-lived ectotherms. Therefore, we consider it relevant to maintain traditional agricultural areas in Mediterranean landscapes, especially in ecotone areas associated with pine forests (generating mosaics with open habitats).

Shorter telomeres are associated with shell anomalies in a long‐lived tortoise

AbstractAge‐related telomere length (TL) variation is relatively well‐described for mammals, birds and other model organisms. Nevertheless, it remains largely unknown in ectotherms, especially turtles and tortoises, which are extremely long‐lived species with slow or negligible senescence. In this study, we described TL dynamics in wild spur‐thighed tortoises (Testudo graeca), one of the chelonian species with the lowest aging rates. By combining cross‐sectional (single) and longitudinal (capture–recapture) samplings, we assessed the relationship between TL and individual characteristics (sex, age, individual growth rate, body condition index, presence of shell anomalies). We did not find any association between TL and sex, individual growth rate, or body condition. However, the relationship with age remains uncertain, likely due to the complex dynamics of TL over time. Interestingly, shorter telomeres correlated significantly with shell anomalies, which are usually assumed as a fitness proxy for reptiles. Overall, our results suggest TL as a potential indicator for ontogenetic studies on tortoises, while its utility as a marker of biological age appears limited.

A comprehensive exploration of diverse skin cell types in the limb of the desert tortoise (Testudo graeca) through light, transmission, scanning electron microscopy, and immunofluorescence techniques

The Greek tortoise, inhabiting harsh desert environments, provides a compelling case for investigating skin adaptations to extreme conditions. We have utilized light microscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and immunofluorescence analysis to describe the structure of the arid-adapted limb skin in the Greek tortoise. Our aim was to identify the cell types that reflect the skin adaptation of this tortoise to arid conditions. Utilizing seven antibodies, we localized and elucidated the functions of various skin cells, shedding light on how the tortoise adapts to adverse environmental conditions. Our findings unveiled numerous scales on the limbs, varying in size and color, acting as protective armor against abrasions, bites, and other potential threats in their rocky habitats. The epidermis comprises four layers: stratum basalis, stratum spinosum, peri-corneous layer, and stratum corneum. Cytokeratin 14 (CK14) was explicitly detected in the basal layer of the epidermis, suggesting a role in maintaining epidermal integrity and cellular function. Langerhans cells were observed between epidermal cells filled with ribosomes and Birbeck granules. Numerous dendritic-shaped Langerhans cells revealed through E-Cadherin signify strong immunity in tortoises' skin. Melanophores were identified using the Melan-A antibody, labeling the cytoplasm, and the SOX10 antibody, labeling the nucleus, providing comprehensive insights into melanophores morphology and distribution. Two types of melanophores were found: dendritic below the stratum basalis of the epidermis and clustered oval melanophores in the deep dermal layer. Varied melanophores distribution resulted in a spotted skin pattern, potentially offering adaptive camouflage and protection against environmental challenges. Numerous myofibroblasts were discerned through alpha-smooth actin (α-SMA) expression, indicating that the Greek tortoise's skin possesses a robust tissue repair and remodeling capacity. B-cell lymphocytes detected via CD20 immunostaining exhibited sporadic distribution in the dermis, concentrating in lymphoid aggregates and around vessels, implying potential roles in local immune responses and inflammation modulation. Employing Tom20 to identify skin cells with abundant mitochondria revealed a notable presence in melanophores and the basal layer of the epidermis, suggesting high metabolic activity in these cell types and potentially influencing cellular functions. These findings contribute to our comprehension of tortoise skin anatomy and physiology, offering insights into the remarkable adaptations of this species finely tuned to their specific environmental habitats.

Body size variation in a lineage of spur-thighed tortoises (Testudo graeca whitei) contrasts with that expected from the species level

Abstract Ectotherms exhibit varying geographic size patterns shaped by environmental and evolutionary factors. This variability is noticeable within taxonomic groups. For instance, certain testudinids follow Bergmann’s rule (body size increases with latitude) and Rensch’s rule (sexual size dimorphism correlates with body size), while others do not. Here we hypothesize that body size patterns can even vary within a monophyletic lineage. To address this, we evaluated the body size patterns of the spur-thighed tortoise Testudo graeca that globally follows Bergmann’s and Rensch’s rules. We specifically investigated the influence of climate variables, latitude and elevation within the subspecies T. g. whitei throughout its natural distribution in North Africa, and in a recently expanded range in SE Spain (20 kya old). We found that males were smaller than females in both regions. The tortoises from SE Spain were smaller than those from North Africa, which showcased the smallest sizes ever reported for the species. Latitude was the main variable to explain tortoise body size. In particular, body size decreased with latitude in both regions, which contrasts with Bergmann’s rule expectations based on species-level findings. Finally, to further contradict species-level expectations, we did not find any statistical correlation between sexual size dimorphism and body size across the two studied regions. Such contradictory outcomes reveal complex geographic size patterns within T. graeca and raise conservation questions about demographic viability at smaller-sized sites.

Evaluation of changes in serum biochemical parameters in zarudny’s spur-thighed tortoises (Testudo graeca zarudnyi) influenced by gender and season

BackgroundThe Zarudny’s spur-thighed tortoise or Iranian tortoise (Testudo graeca zarudnyi) has just been reported from Iran so far. Therefore, the purpose of this study was to determine the effects of season and gender on serum biochemical parameters of this valuable species in Iran.ResultsThis study was performed on 20 clinically healthy adult Zarudny’s spur-thighed tortoises. Blood samples were collected from the jugular vein and then serum biochemical parameters and body weight were measured in autumn and winter. The following biochemical parameters were measured: total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein, total protein, creatinine, urea, glucose, calcium, inorganic phosphorus, total bilirubin, uric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase. Urea concentration increased significantly at emergence from hibernation. Also, in winter, total protein, phosphorous, creatinine, total bilirubin concentrations and alkaline phosphatase activity decreased significantly compared to autumn, but aspartate aminotransferase and lactate dehydrogenase activities were significantly higher than in autumn. There was no significant difference for the parameters mentioned above by gender, except for phosphorous. Phosphorous concentration in females was significantly higher than that of males in autumn. Cholesterol and low-density lipoprotein cholesterol levels in females were significantly higher than males in autumn and winter.ConclusionsThe present study is the first study that monitors the serum biochemical parameters of adult Zarudny’s spur-thighed tortoises based on season and gender. Seasonal reference intervals should be used for biochemical parameters in this valuable species. Also, sex-specific reference intervals for phosphate and cholesterol are necessary.

Taking advantage of reference-guided assembly in a slowly-evolving lineage: Application to Testudo graeca.

Obtaining de novo chromosome-level genome assemblies greatly enhances conservation and evolutionary biology studies. For many research teams, long-read sequencing technologies (that produce highly contiguous assemblies) remain unaffordable or unpractical. For the groups that display high synteny conservation, these limitations can be overcome by a reference-guided assembly using a close relative genome. Among chelonians, tortoises (Testudinidae) are considered one of the most endangered taxa, which calls for more genomic resources. Here we make the most of high synteny conservation in chelonians to produce the first chromosome-level genome assembly of the genus Testudo with one of the most iconic tortoise species in the Mediterranean basin: Testudo graeca. We used high-quality, paired-end Illumina sequences to build a reference-guided assembly with the chromosome-level reference of Gopherus evgoodei. We reconstructed a 2.29 Gb haploid genome with a scaffold N50 of 107.598 Mb and 5.37% gaps. We sequenced 25,998 protein-coding genes, and identified 41.2% of the assembly as repeats. Demographic history reconstruction based on the genome revealed two events (population decline and recovery) that were consistent with previously suggested phylogeographic patterns for the species. This outlines the value of such reference-guided assemblies for phylogeographic studies. Our results highlight the value of using close relatives to produce de novo draft assemblies in species where such resources are unavailable. Our annotated genome of T. graeca paves the way to delve deeper into the species' evolutionary history and provides a valuable resource to enhance direct conservation efforts on their threatened populations.

Sexual Dimorphism in <i>Testudo graeca zarudnyi</i> (Testudines: Testudinidae) from Yazd Province, Central Iran

To investigate morphological traits of the poorly known Zarudny’s tortoise, Testudo graeca zarudnyi Nikolsky, 1896, which has only been reported from Iran so far, we have conducted morphometric analysis to test sexual differences (sexual size and shape dimorphism, SSD) in Marvar Prohibited Hunting Area (central Iran). Our findings showed that males have wider carapace and larger hind limbs; while, females have longer plastron. This pattern has resulted in shape differences with males; that is, have more domed and wider carapace than to the females. Generally, Females are larger than males we will discuss it as a force to fecundity selection. The results showed that this subspecies does not follow Rench’s rule which argues that males are larger than females.

First molecular detection of Francisella tularensis in turtle (Testudo graeca) and ticks (Hyalomma aegyptium) in Northwest of Iran

Francisella tularensis, causative agent of tularemia, is a contagious zoonotic ailment. This study was aimed to molecularly detect F. tularensis in tortoise blood (n = 100) and ticks (n = 100) collected in the West Azerbaijan province, Iran suing a 16SrRNA gene of the Francisella genus through employment of the Nested-PCR technique. The identified ticks were s Hyalomma aegyptium by morphological analysis. Seven percent (with a 95% CI: 3.5%–13.75%) of animal blood samples yielded positive results for the presence of the Francisella. Meanwhile, the Francisella was identified in tick samples at a rate of fifteen percent (15%) (with a 95% CI: 9%–23%). The samples containing positive results were specifically classified as F. tularensis subsp. holarctica. The samples were taken from ticks belonging to the H. aegyptium species that were gathered in Oshnavieh, southern part of West Azerbaijan province, Iran. This research was aimed to validate the existence of F. tularensis in ticks found within the West Azerbaijan province. Consequently, it is vital to acknowledge the potential of these ticks to transmit the bacteria to both livestock and humans through tick bites in this specific area.

Scavenging by young tortoises (Testudo sp.) could induce their predation by the Eurasian Eagle-Owl (Bubo bubo)

Abstract Eating owl pellets by non-predator vertebrates is a rare and difficult-to-prove food chain relationship. In this paper, I reported the first record of a Spur-thighed Tortoise (Testudo graeca) eating a Eurasian Eagle-Owl (Bubo bubo) pellet with remains of a right Wood Pigeon (Columba palumbus) wing. Scavenging of food remains around owl nests by young tortoises may possibly explain the few cases of tortoise-eating Eurasian Eagle-Owls.

Tick infestation in spur-thighed tortoise population: a pilot study for unraveling epidemiological patterns and demographic consequences

Ectoparasites, such as ticks, modulate host population dynamics by impacting demographic traits. They transmit infectious agents among their hosts, posing a critical threat to animal and public health. This study aimed to characterize and analyze the Hyalomma aegyptium infestation on one of its main hosts, the spur-thighed tortoise, its effects on demographic traits, and to determine the diversity of infectious agents present in both ticks and tortoises in the Maamora forest (northwestern Morocco). Our results show that 100% of the tortoises were parasitized by adult ticks in spring, an infestation intensity of 4 ticks/tortoise (5.1 and 3.6 ticks/tortoise in males and females, respectively; 4.2 and 3.3 ticks/tortoise in gravid and non-gravid females, respectively) and an abundance ranging from 1 to 12. Although without significant differences, male tortoises had higher tick abundances than females. The interaction of tortoise sex and body condition was significantly related to tick abundance, male body condition decreased with higher tick abundance in contrast to females. Nevertheless, the interaction of body condition and reproductive stage of females was not significantly related to tick abundance. Gravid females were significantly associated with tick abundance, showing a slightly higher infestation than non-gravid females. Molecular analysis of pooled tick samples revealed the presence of Ehrlichia ewingii, Candidatus Midichloria mitochondrii, and Rickettsia africae, with a minimum infection rate of 0.61 to 1.84%. However, blood sample analysis of the tortoises was infectious agent-free, pinpointing a lack of significant health problems. Given the possible effect on the transmission of zoonotic diseases by spur-thighed tortoises associated with their frequent collection as pets, it should be surveyed to control possible human health problems. In conservation terms, as a long-lived species, the role of tick infestation in demographic traits might be included in the management and conservation programs of spur-thighed tortoises.

Molecular detection of Coxiella burnetii in blood samples of turtles (Testudo graeca) and their associated ticks

Molecular detection of Coxiella burnetii in blood samples of turtles (Testudo graeca) and their associated ticks

Data on the largest specimens of Testudo graeca ibera Pallas, 1814 found in Bulgaria with five new records

Data on the largest specimens of Testudo graeca ibera Pallas, 1814 found in Bulgaria with five new records

A Retrospective Study on Wild Animals Admitted to Animal Rescue and Rehabilitation Centres in Türkiye*

The aim of this study was to retrospectively reveal the rehabilitation numbers of wild animals admitted to rescue and rehabilitation centres in Türkiye between 2017 and 2021, to investigate the wild animals hospitalized in the centres under mammal, bird and reptile classes at the level of order and species, and to identify the deficiencies in wildlife rehabilitation and to make recommendations. The main material of the research was the data for the years 2017-2021 obtained from the Wildlife Information System-YABIS, a database of the Ministry of Agriculture and Forestry. As the method, numerical data on the species downloaded from YABIS were analysed. Accordingly, it was understood that a total of 35764 cases were admitted to all rescue centres across Türkiye. About 61% of these cases were treated and released into nature, about 34% died and about 5% were placed in zoos. While the Cetartiodactyla was the most affected order in mammals, the most accepted species in the centres was the Roe deer (Capreolus capreolus). In birds, the Columbiformes was the most affected order, the most accepted species in the centers was the Rock dove (Columba livia). In reptiles, the Testudinata was the most affected order, the most accepted species in the centres was the Mediterranean spur-thighed tortoise (Testudo graeca). The increase in the number of cases brought to rehabilitation centers between 2017-2021, excluding 2019, clearly showed the role of rescue and rehabilitation centers in protecting wild animals and providing sustainable wildlife.

Ultrasonography and CT examination of ovarian follicular development in 'Testudo graeca' during 1 year in captivity.

The Greek or Mediterranean tortoise (Testudo graeca), commonly known as the spur-thighed tortoise, is a species of tortoise in the family Testudinidae that is found in North Africa, Western Asia and Europe. Several species of this genus are under threat in the wild, mainly from habitat destruction therefore, accurate information about the sexual cycle and the exact time of follicular development can be effective in successful reproduction planning and preventing species extinction. The experimental part of this study is focused on the evaluation of the reproductive apparatus of tortoises and development of follicles in ovaries during 1 year by ultrasonography and computed tomography. Ultrasonography and CT scanning was carried out on 10 female 'T. Graeca' tortoises which were maintained at Tehran Eram Zoo from November 2021 to November 2022 (1 year). Ultrasonography and CT exams were performed on all animals. In the first study, on 28 November 2021, the ovarian cycle was characterized by the presence of previtellogenic follicles and atretic follicles. The numbers of follicles were best demonstrated on CT examination. Ultrasonography and CT scan in the second study showed no change in type of follicles from previous study. In the third study, atretic follicles were more than previtellogenic follicles, whereas in the fourth one, the numbers of previtellogenic follicles were more than atretic follicles. In following, at 6 and 29 June 2022, the absence of atretic follicles and just presence of previtellogenic follicles were observed. On 20 July 2022, the presence of a few number of atretic follicles and more previtellogenic follicles were noticed. On 30 July 2022, the absence of atretic follicles and presence of previtellogenic follicles and, for the first time, preovulatory follicles were obvious. In four following studies the absence of atretic follicles and the presence of previtellogenic follicles were diagnosed. In the last study, the absence of atretic follicles and the presence of preovulatory and previtellogenic follicles were observed. In current study, the reproductive cycle of female T. Graeca is evaluated during 1 year in captivity with ultrasonography and CT scan. It can be concluded that in captivity, the reproductive cycle does not result in the formation of eggs or calcareous shells and atretic follicles in the study year continued as before; so this should be highly considered in captivity breeding programmes. This study also revealed that among imaging modalities, CT scan is the best modality for detecting the shape, size, type and numbers of the follicles for further evaluation of location and shape of the follicles.