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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.

Simple Summary: This study explored the morphological differences in the carapaces of three tortoise species—Leopard tortoises (Stigmochelys pardalis), African spurred tortoises (Centrochelys sulcata), and Greek tortoises (Testudo graeca)—raised in captivity in Türkiye. Using 3D scanning and geometric morphometric analysis, we identified the vital shape variations and dimensional features across species. Principal component analysis revealed that PC1 and PC3 were significant in distinguishing species, with Leopard tortoises showing higher carapace shapes and African spurred tortoises having shorter carapaces. An allometric effect indicated that smaller tortoises had higher shells. These findings provide valuable insights for taxonomy and clinical practice, emphasizing the need to consider shape variations in tortoise care and species identification. 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. [ABSTRACT FROM AUTHOR]

Tick-borne pathogens are a worldwide threat to public health that can only be mitigated by knowledge on tick-host associations coupled with surveillance of their infection by pathogenic microorganisms. This information is not equally available throughout tick vector distribution range and is deficient in some geographical areas. In this study we did a molecular survey of tick-borne pathogens associated with different tick species in Morocco. We analyzed four different species of Hyalomma (Hyalomma aegyptium, Hyalomma anatolicum, Hyalomma dromedarii and Hyalomma impeltatum) and Rhipicephalus sanguineus sensu lato ticks from four vertebrate hosts and from the vegetation. The most common tick species collected was H. aegyptium feeding on Testudo graeca tortoises, followed by H. anatolicum from the vegetation. Hyalomma aegyptium feeding on T. graeca was found infected with Borrelia turcica, representing the first detection for Western North Africa, and Rickettsia sibirica mongolitimonae, also detected for the first time in Morocco. Rickettsia aeschlimannii was also detected in H. aegyptium feeding on T. graeca. Additionally, R. aeschlimannii was detected in H. anatolicum from the vegetation, and Rickettsia massiliae in R. sanguineus s.l. from an Algerian hedgehog Atelix algirus. Because H. aegyptium are common parasites of T. graeca tortoises, and these animals are subjected to pet trade, it is important to evaluate the associated human health risks through regular surveillance and perform awareness campaigns for prevention of the general public.

Age‐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. [ABSTRACT FROM AUTHOR]

Context: Passive habitat rewilding after rural abandonment can affect wildlife differently depending on the type of habitats that it generates. Objective: Evaluate 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. Methods: We 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). Results: The 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. Conclusions: Our 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). [ABSTRACT FROM AUTHOR]

Abstract Background 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. Objective 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. Methods 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. Results 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. Conclusion 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.

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: 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. Objective: 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. Methods: 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. Results: 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. Conclusion: 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. [ABSTRACT FROM AUTHOR]

Recovery time from anesthesia can be extended in reptiles, consequently patients undergoing general anesthesia, require prolonged monitoring period which increases the probability of postoperative complications. Therefore, prolonged recovery time following inhalant anesthesia is a common complication in chelonians. Ability to intracardiac shunting and bypassing blood from pulmonary circulation, may contribute to their unpredictable inhalant anesthetic recovery times. The acupuncture point Governing vessel (GV-26) has been demonstrated to reduce anesthetic recovery times from inhalant anesthesia in many species. In this study eight spur-thighed tortoises (Testudo gracea) were anesthetized by isoflurane administration for 60 min. The trial was performed in two parts with four weeks washout period. In the first time, once isoflurane administration was discontinued, the tortoises received no therapy in the recovery period and in the second part of the study, tortoises received GV-26 electroacupuncture stimulation. Physiologic variables, anesthetic parameters, time to first movement and time to extubation were recorded. Data were compared with the use of independent sample t tests. Tortoises receiving GV-26 electroacupuncture had a significantly reduced time to return of voluntary movement (P

Recovery time from anesthesia can be extended in reptiles, consequently patients undergoing general anesthesia, require prolonged monitoring period which increases the probability of postoperative complications. Therefore, prolonged recovery time following inhalant anesthesia is a common complication in chelonians. Ability to intracardiac shunting and bypassing blood from pulmonary circulation, may contribute to their unpredictable inhalant anesthetic recovery times. The acupuncture point Governing vessel (GV-26) has been demonstrated to reduce anesthetic recovery times from inhalant anesthesia in many species. In this study eight spur-thighed tortoises (Testudo gracea) were anesthetized by isoflurane administration for 60 min. The trial was performed in two parts with four weeks washout period. In the first time, once isoflurane administration was discontinued, the tortoises received no therapy in the recovery period and in the second part of the study, tortoises received GV-26 electroacupuncture stimulation. Physiologic variables, anesthetic parameters, time to first movement and time to extubation were recorded. Data were compared with the use of independent sample t tests. Tortoises receiving GV-26 electroacupuncture had a significantly reduced time to return of voluntary movement (P<0.001), and a significantly reduced time to extubation (P<0.001). Furthermore, time from first movement to extubation was significantly shorter (P<0.05). Hence, the use of GV)26 electroacupuncture results in significant reduction of anesthetic recovery time in spur-thighed tortoises which have received inhalant anesthetic. [ABSTRACT FROM AUTHOR]

Tortoises of the genus Testudo are the main hosts of Hyalomma aegyptium ticks. This species serves as a vector of several zoonotic pathogens. Therefore, the present study aimed to investigate the presence of four pathogens associated with H. aegyptium ticks obtained from tortoises from Tunisia. Conventional, multiplex and nested PCRs were used for Aanaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii and Babesia spp. screening. The molecular analyses revealed the presence of A. phagocytophilum and Babesia spp. None of the ticks, were infected by E. canis or C. burnetii species. Co-infection was detected in four ticks. As a conclusion, this is the first detection of A. phagocytophilum and Babesia spp. in H. aegyptium ticks collected from tortoises, in Tunisia. Thus, considering these results, the spur-thighed tortoise constitute a potential host of H. aegyptium which plays an important role in the transmission of pathogenic agents affecting both human and animals. In term of public health, a strict control and surveillance should be carried to reduce the circulation of such pathogens between different hosts. [ABSTRACT FROM AUTHOR]

Simple Summary: Assessing the body size and body shape variations between sexes and geographical populations can help us understand the adaptive responses of organisms in the face of the pressures to which they are subjected. To evaluate the influence of habitat-type conditions, we selected six Algerian populations of Testudo graeca living in different environments. The results of the traditional morphometric analyses showed that body size and shell shape were smaller and flattened, respectively, in males, especially under unfavorable conditions for tortoises; these changes were jointly caused by anthropogenic and natural pressures. We found clear evidence in several tortoise species that differences in growth durations up to the onset of maturity resulted primarily in different sizes at maturity and ultimately in different adult sizes. Using data for the body size and shell shape of Algerian Testudo graeca, we assessed how proximate causes shaped the observed variation in the morphology of adults. All of the studied populations displayed significant sexual size and shape dimorphisms. Relative to body length, females displayed larger, more voluminous and domed shells than males. We found clear evidence that variation in body size at maturity influenced sexual size dimorphism. Body size at maturity depends on the duration of growth from hatching up to the point of reaching sexual maturity. In the studied populations, sexual maturity, estimated by counting growth lines, was always reached earlier in males than in females (a time difference of 1.4–3.0 years). Similar to sexual size dimorphism, geographic variation in adult body sizes was also influenced by variations in the corresponding sizes at maturity. Remarkably, the population with the largest tortoises had the latest mean maturation time: 9.1 for males and 10.5 for females. Thus, the later completion of maturation was a determinant for a larger size in adulthood. The largest tortoises among the studied populations were measured at the Djelfa locality, where the recorded sizes of males and females reached 186 and 230 mm, respectively. [ABSTRACT FROM AUTHOR]

Context: Land-use change is one of the main threats to biodiversity on the global scale. Legacy effects of historical land-use changes may affect population dynamics of long-lived species, but they are difficult to evaluate through observational studies alone. We present here an interdisciplinary modelling approach as an alternative to address this problem in landscape ecology. Objectives: Assess effects of agricultural abandonment and anthropisation on the population dynamics of long-lived species. Specifically, we evaluated: (a) how changes in movement patterns caused by land-use change might impact population dynamics; (b) time-lag responses of demographic variables in relation to land-use changes. Methods: We applied an individual-based and spatial-explicit simulation model of the spur-tighed tortoise (Testudo graeca), an endangered species, to sequences of real-world landscape changes representing agricultural abandonment and anthropisation at the local scale. We analysed different demographic variables and compared an "impact scenario" (i.e., historical land-use changes) with a "control scenario" (no land-use changes). Results: While agricultural abandonment did not lead to relevant changes in demographic variables, anthropisation negatively affected the reproductive rate, population density and the extinction probability with time-lag responses of 20, 30 and 130 years, respectively, and caused an extinction debt of 22%. Conclusions: We provide an understanding of how changes in animal movement driven by land-use changes can translate into lagged impacts on demography and, ultimately, on population viability. Implementation of proactive mitigation management are needed to promote landscape connectivity, especially for long-lived species for which first signatures of an extinction debt may arise only after decades. [ABSTRACT FROM AUTHOR]

Crimean-Congo haemorrhagic fever is endemic in Bulgaria where the primary vector and reservoir are ticks Hyalomma marginatum. The present study examined the role of ticks Hyalomma aegyptium parasitizing on tortoises Testudo graeca in circulation of Crimean-Congo haemorrhagic fever virus. A total of 81 ticks H. aegyptium were collected from 23 specimens of T. graeca in Kardzhali District, SE Bulgaria. Real-time RT-PCR was performed for the detection of CCHFV RNA. CCHFV genome was detected in 10 specimens of H. aegyptium collected from two T. graeca hosts (infection rate 12.3 %). Further studies on H. aegyptium and their hosts are needed to assess their role as CCHFV vectors and reservoirs in SE Bulgaria. [ABSTRACT FROM AUTHOR]

Thyroid hormones and the factors influencing them are rarely studied in tortoises. This study therefore aimed to calculate reference intervals (RI) for thyroid hormones and iodine concentrations by species, sexes, and seasons in four species of adult Mediterranean tortoises (Testudo spp.) and to evaluate possible correlations between plasma thyroid hormones, serum iodine, plasma protein, and increased liver and kidney values. Thyroid hormones (total tetraiodothyronine [tT4], free tetraiodothyronine [fT4], total triiodothyronine [tT3], and free triiodothyronine [fT3]) were measured in plasma from adult, healthy Hermann's (Testudo hermanni, n = 255), spur-thighed (Testudo graeca, n = 89), marginated (Testudo marginata, n = 72), and Russian tortoises (Testudo horsfieldii, n = 30). Species, sex, and seasonal variations were determined by Bayesian information criterion (BIC), and correlations between thyroid hormones, protein, iodine, and increased liver/kidney values were evaluated by Spearman's rank correlation test. Total T4 did not reveal any species, sex, or seasonal differences (RI: 0.102–0.455 µg/dl), while seasonal differences were found for fT4 (RI: spring 0.624–9.012; summer 0.379–5.476; fall 0.376–5.426 pmol/L). The tT3 concentrations differed significantly depending on species, season, and the interaction of species × season. Seasonal differences were also found for fT3 and iodine. Several significant (P ≤ 0.05) correlations were detected between the analytes, including a positive correlation between tT4 and fT4. These results provide a tool for the evaluation of thyroid hormone concentrations in Mediterranean tortoises and confirm the influence of season on thyroid function in these animals. [ABSTRACT FROM AUTHOR]

The spur-thighed tortoise (Testudo graeca L., 1758) is classified as a vulnerable species by the IUCN. In this context, it is important to explore the factors affecting its populations to set up an effective conservation plan. The present study was carried out in seven localities of Morocco with the objective to determine the ticks parasitizing this tortoise and their prevalence in relation to host characteristics and environmental factors. We examined 392 free-living tortoises for the presence of ticks during the spring 2018. A total number of 2238 adult ticks (no immature tick detected) were collected and all identified as Hyalomma aegyptium. Infection with at least one tick was detected in 77.6% ± 6.5 of the total tortoises. The infestation prevalence varied according to localities. Infestation intensity varied from 1.5 to 12, while tick abundance ranged from 0.65 to 12 ticks/tortoise. Male tortoises were more infested than females (P < 0.01) and the infestation was age-dependant. Our statistical result provides that the best model drivers tick parasitism was which included locality and age. Further studies are needed to assess the impact of the high intensity of H. aegyptium infestation on the tortoise's health status and to assess the potential transmission of zoonotic pathogens. [ABSTRACT FROM AUTHOR]

Seven nematodes collected from a crushed turtle were brought to Selçuk University Faculty of Veterinary Parasitology Department. A few of them were transparented in lactophenol approximately for three weeks. After the parasites were cleared, their head regions were examined microscopically in apical and lateral positions. The posterior ends of the samples were also examined to separate male and females. According to the results of the microscopic examinations, it was found that five of seven parasites were female and remain two were male and parasites were identified as Angusticaecum holopterum (Rudolphi, 1819) considering their morphological characteristics.

The study of the distribution of protected animal species in Europe is especially relevant in a changing climate. Therefore, in this work, we tried to solve the problem of the possibility of habitation of tortoises Testudo graeca Linnaeus, 1758 and Testudo hermanni Gmelin, 1789 in Eastern Europe by using species distribution models (SDMs). We used bioclimatic variables from the CliMond dataset (18 uncorrelated variables of 35) and 19 Paleoclim variables of the "early-Holocene" and "mid-Holocene". Packages Maxent and 'ntbox' were employed. In addition to our data, we used findings listed in the GBIF databases: 1,935 points for T. graeca and 991 points for T. hermanni. It has been shown that subspecies of tortoises differ in the characteristics of the ecological niche. In addition to direct anthropogenic influences, the limiting factor is the "Mean temperature of coldest quarter" (bio11) for both species. Moreover, T. graeca is less demanding and can tolerate both frost and higher temperatures during drier periods than T. hermanni. Modelling found that in the future it is possible for these species will expand in a north-eastern direction, where potentially suitable habitats will appear: by 2090 in the South of Ukraine (Odesa Region, Crimea) and East Ukraine (floodplain of the Siversky Donets River of the Don basin). [ABSTRACT FROM AUTHOR]

Animal populations have developed multiple strategies to deal with environmental change. Among them, the demographic buffering strategy consists in constraining the temporal variation of the vital rate(s) that most affect(s) the overall performance of the population. Tortoises are known to buffer their temporal variation in adult survival, which typically has the highest contribution to the population growth rate λ, at the expense of a high variability on reproductive rates, which contribute far less to λ. To identify the effects of projected increases in droughts in its natural habitat, we use field data collected across 15 locations of Testudo graeca in southeast Spain over a decade. We analyse the effects of environmental variables on reproduction rates. In addition, we couple the demographic and environmental data to parameterise an integral projection model to simulate the effects of different scenarios of drought recurrence on λ under different degrees of intensity in the survival–reproduction tradeoff. We find that droughts negatively affect the probability of laying eggs; however, the overall effects on λ under the current drought recurrence (one/decade) are negligible when survival is constant (independent of the reduction of reproduction by drought events) and when survival increased as a tradeoff with the reduction of reproduction rates, with a threshold to population viability at three or more droughts/decade. Additionally, we show that, although some species may buffer current environmental regimes by carefully orchestrating how their vital rates vary through time, a demographic buffering strategy is insufficient to ensure population viability in extreme regimes. Our findings support the hypothesis that the demographic buffering strategy has a limit of effectiveness when adverse conditions occur frequently. Our methodological approach provides a framework for ecologists to determine how effective the management of environmental drivers can be for demographically buffering populations, and which scenarios may not provide long‐term population persistence. [ABSTRACT FROM AUTHOR]

Using data for the body size and shell shape of Algerian Testudo graeca, we assessed how proximate causes shaped the observed variation in the morphology of adults. All of the studied populations displayed significant sexual size and shape dimorphisms. Relative to body length, females displayed larger, more voluminous and domed shells than males. We found clear evidence that variation in body size at maturity influenced sexual size dimorphism. Body size at maturity depends on the duration of growth from hatching up to the point of reaching sexual maturity. In the studied populations, sexual maturity, estimated by counting growth lines, was always reached earlier in males than in females (a time difference of 1.4–3.0 years). Similar to sexual size dimorphism, geographic variation in adult body sizes was also influenced by variations in the corresponding sizes at maturity. Remarkably, the population with the largest tortoises had the latest mean maturation time: 9.1 for males and 10.5 for females. Thus, the later completion of maturation was a determinant for a larger size in adulthood. The largest tortoises among the studied populations were measured at the Djelfa locality, where the recorded sizes of males and females reached 186 and 230 mm, respectively.

A cycle of glacial and interglacial periods in the Quaternary caused species' ranges to expand and contract in response to climatic and environmental changes. During interglacial periods, many species expanded their distribution ranges from refugia into higher elevations and latitudes. In the present work, we projected the responses of the five lineages of Testudo graeca in the Middle East and Transcaucasia as the climate shifted from the Last Glacial Maximum (LGM, Mid - Holocene), to the present. Under the past LGM and Mid-Holocene bioclimatic conditions, models predicted relatively more suitable habitats for some of the lineages. The most significant bioclimatic variables in predicting the present and past potential distribution of clades are the precipitation of the warmest quarter for T. g. armeniaca (95.8 %), precipitation seasonality for T. g. buxtoni (85.0 %), minimum temperature of the coldest month for T. g. ibera (75.4 %), precipitation of the coldest quarter for T. g. terrestris (34.1 %), and the mean temperature of the driest quarter for T. g. zarudyni (88.8 %). Since the LGM, we hypothesise that the ranges of lineages have either expanded (T. g. ibera), contracted (T. g. zarudnyi) or remained stable (T. g. terrestris), and for other two taxa (T. g. armeniaca and T. g. buxtoni) the pattern remains unclear. Our analysis predicts multiple refugia for Testudo during the LGM and supports previous hypotheses about high lineage richness in Anatolia resulting from secondary contact. [ABSTRACT FROM AUTHOR]

Mate searching is a key component of sexual reproduction that can have important implications for population viability, especially for the mate‐finding Allee effect. Interannual sperm storage by females may be an adaptation that potentially attenuates mate limitation, but the demographic consequences of this functional trait have not been studied. Our goal is to assess the effect of female sperm storage durability on the strength of the mate‐finding Allee effect and the viability of populations subject to low population density and habitat alteration. We used an individual‐based simulation model that incorporates realistic representations of the demographic and spatial processes of our model species, the spur‐thighed tortoise (Testudo graeca). This allowed for a detailed assessment of reproductive rates, population growth rates, and extinction probabilities. We also studied the relationship between the number of reproductive males and the reproductive rates for scenarios combining different levels of sperm storage durability, initial population density, and landscape alteration. Our results showed that simulated populations parameterized with the field‐observed demographic rates collapsed for short sperm storage durability, but were viable for a durability of one year or longer. In contrast, the simulated populations with a low initial density were only viable in human‐altered landscapes for sperm storage durability of 4 years. We find that sperm storage is an effective mechanism that can reduce the strength of the mate‐finding Allee effect and contribute to the persistence of low‐density populations. Our study highlights the key role of sperm storage in the dynamics of species with limited movement ability to facilitate reproduction in patchy landscapes or during population expansion. This study represents the first quantification of the effect of sperm storage durability on population dynamics in different landscapes and population scenarios. [ABSTRACT FROM AUTHOR]

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. • Dendritic-shaped Langerhans cells were identified by E-Cadherin that indicated robust immunity in tortoise skin. • Melanocytes were identified by Melan-A antibody, which labeled the cytoplasm, and the Sox10 antibody, which labeled the nucleus. • Dendritic melanocytes were discovered under the epidermis, and clustered melanocytes were discovered in the dermal layer. • Several myofibroblasts were identified using alpha-smooth actin. B-cell lymphocytes were found in the dermis. • Tom20 identified skin cells with numerous mitochondria in melanocytes and the epidermis' basal layer. [ABSTRACT FROM AUTHOR]

Zoo and wildlife practitioners are constantly exposed to persistent dangers during physical restraint of wild animals. Chemical immobilization in reptiles is unpredictable due to their ectothermic nature. This study aims to determine a safe and effective anesthetic protocol for immobilization in chelonians and other reptiles. Varied doses of diazepam ketamine combinations were administered and evaluated in 16 healthy land tortoises (Testudo graeca). The tortoises were divided into four groups (4 per group) labeled DK-1 through DK-4. DK-1 had a combination of 44mg/kg of ketamine with 0.25mg/kg of diazepam. DK-2 had 22mg/kg of ketamine with 0.25mg/kg of diazepam. DK-3 had 44mg/kg of ketamine with 0.5mg/kg diazepam while DK-4 had 22mg/kg of ketamine with 0.5mg/kg of diazepam. Anesthetic effects were monitored to determine the duration required for partial extension of the head and limbs, full extension of limbs, and complete recovery from anesthesia. These visual inspection of partial recovery and full extension were adapted as surface and deep anaesthesia respectively in describing the depth of anaesthesia. All the time intervals were recorded in minutes, and summarized as mean and standard deviation. ANOVA was used to test for significance across the groups. Full extension of head and limbs was achieved within mean periods of 10, 15.5 and 13 minutes in DK-1, DK-3 and DK-4 respectively. Tortoises in DK-2, which were only sedated, demonstrated only surface depth of anaesthesia. Complete recovery occurred in mean periods of 128, 25, 158 (p < 0.05) and 132 minutes for groups DK-1, DK-2, DK-3, and DK-4 respectively. Diazepam-Ketamine anaesthetic cocktail provides a safe protocol for chemical restraint in tortoises. A higher dose of diazepam produced a longer duration of complete recovery. [ABSTRACT FROM AUTHOR]

Abstract Macroparasites in general, and ectoparasites in particular, have the potential to regulate host population dynamics. In this context, this study addresses the tick parasitism traits of the Mediterranean spur-thighed tortoise (Testudo graeca) in the core area of its distribution range (northwestern Morocco, Maamora). It was discovered that 92.5% of the tortoises were parasitized by ticks in spring, with an infestation intensity and an abundance of 6.7 and 6.2 tick/tortoise, respectively. The observed parasitization rates were among the highest reported worldwide for T. graeca , which could relate to density-depended effects driving host-parasite interactions. The main tick species that parasitized the tortoises were Hyalomma aegyptium (95.6% of the ticks and in the 100% of the parasitized tortoises), Hy. marginatum , Hy. excavatum and Hy. scupense. Individual predictors for the tortoises, such as age, sex and the interaction between body condition and sex, were significantly related to tick abundance. Age-related behavioural differences might favour a higher host-tick effective contact in adults than in juveniles. The fact that males are more active in spring - the breeding season - might explain the observed male-bias in tick abundance and may also be responsible for the negative effect of male body condition on tick infestation rate in contrast to females. Given the potential role played by parasites as regards modulating population dynamics, our results suggest that ticks should be taken into account in the conservation and management programmes of this tortoise species. [ABSTRACT FROM AUTHOR]

Long-lived species are particularly interesting for investigation of trade-offs that shape reproductive allocation and the effective contribution to the next generations. Life history theory predicts that these species will buffer environmental stochasticity via changes in the reproductive investment, while maintaining high adult survival rates. The spur-thighed tortoise was selected as a case study in order to investigate the relationship between the linked maternal characteristics (size and age) and related traits in their hatchlings. We tracked naturally emerging hatchlings from young and old females under semi-natural conditions to test variations in hatchling numbers, body mass, size and survival over two years. We used linear mixed-effect models to analyze variations in hatchling body mass and size, and a mark–release–recapture framework to model their survival. Our study illustrates that old females of long-lived species have greater offspring numbers, greater survival and smaller size when compared with those of young females. The interannual variability evidenced the reduced offspring number and survival in the lower autumn rainfall and spring mean temperature year. Our results highlight the role of maternal age and climatic conditions in the population dynamics and the need for long-term studies of reproduction traits for designating adequate conservation strategies.

The trading and collection of wild animals as pets may be cause for concern regarding animal welfare and species conservation. These concerns can be exemplified by Mediterranean spur-thighed tortoise (Testudo graeca), a long-living species whose use as pets is long established. The human dimension plays a major role in the wildlife for the pet collection, and is particularly important in countries like Morocco, where this might pose a threat to the conservation of the species involved. This study, which is based on a questionnaire survey (n = 480 participants), documents the fact that many people in Morocco keep tortoises as pets: 55% of the participants in the survey and over two tortoises/person. Importantly, most captive tortoises, particularly juveniles, had been collected directly from wild populations by their owners (42%, n = 264). In general, the tortoise owners had limited knowledge of their tortoises’ habits and requirements, although rural people were more likely to acknowledge that the tortoise is a wild and threatened species. Our study reveals that non-commercial collection is a common activity in Morocco that may threaten wild tortoise populations and hence species conservation, and it could have consequences regarding the welfare of the animals. We were also able to identify the profile of people towards whom education campaigns should be directed in order to reduce the number of tortoises collected from wild populations. Additional field research should also be conducted to quantify the impact of pet collection on wild tortoise populations.

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Hyalomma aegyptium (Linnaeus, 1758) is a hard-tick species of the Ixodidae family with the main adult hosts being Palearctic tortoises of the genus Testudo. In Morocco, it is commonly found infecting Testudo graeca that has a wide geographic distribution and is subject to protection through international illegal trade legislation. This tortoise tick has been reported as an important vector of various human and animal pathogens. Unfortunately, to date, there are no strategies to control this hematophagous ectoparasite. The present study was conducted in laboratory to evaluate the acaricidal activity of essential oils (EOs) extracted from six plants of Moroccan origin as an alternative against H. aegyptium. All EOs tested displayed toxic effects on different life stages of H. aegyptium. Mentha suaveolens subsp. timija (LC50 = 0.910 µL/mL and LC90 = 1.465 µL/mL) and Satureja calamintha EOs (LC50 = 0.927 µL/mL and LC90 = 1.347 µL/mL) exhibited higher activity against hatching eggs, whereas Chenopodium ambrosioides EO was comparatively more toxic to larvae (LC50 = 0.444 µL/mL and LC90 = 0.918 µL/mL). Juniperus thurifera var. africana (LC50 = 0.0045 mL/cm2 and LC90 = 0.0118 mL/cm2) and Lavandula pedunculata subsp. atlantica EOs (LC50 = 0.0036 mL/cm2 and LC90 = 0.0110 mL/cm2) caused high mortality in nymphs. The effect of different EOs is discussed according to their chemical composition and bioactive components. [ABSTRACT FROM AUTHOR]

In this study, some biochemical parameters (glucose, urea, calcium, triglyceride, cholesterol, creatinine, uric acid, total protein, iron, albumin, phosphorus, H.D.L., magnesium) hemoglobin and hematocrit depending on the pre-reproductive and reproductive periods and sex were determined in a total of 20 Testudo graeca specimens in the Çanakkale (Turkey). For the biochemical parameters, 0,5 ml of blood was collected from the scale in the lateral ligament of the neck with a 5-ml aspirating syringe. The blood was transferred into the biochemical tubes and centrifuged; the plasma part was separated and was measured by spectrophotometer. In conclusion, plasma biochemistry parameters were determined on sexrelated pre-reproduction and reproduction periods in T. graeca. When comparing the pre-reproductive and the reproductive period in terms of glucose, uric acid and iron values, statistical differences were found in the females. It was determined that cholesterol values of males were significantly different between periods. In the females, glucose value was found high in the pre-reproductive period and uric acid and iron values were high in the reproductive period. Cholesterol values of males were found high in the pre-reproductive period. Iron levels in the pre-reproductive period were significantly different between males and females. [ABSTRACT FROM AUTHOR]

A prospective, randomized, crossover experimental pilot study was performed to compare the anesthetic quality and recovery times of intramuscular (IM) ketamine/medetomidine and S-ketamine/medetomidine in Testudo spp. Four adult male Hermann's tortoises (Testudo hermanni) and one adult male spur-thighed tortoise (Testudo graeca) were used in this study. Each animal underwent two different anesthetic protocols twice, with a washout period of two weeks in between each anesthetic event. This resulted in 20 anesthetic procedures. Protocols consisted of 0.1 mg/kg medetomidine IM with either 15 mg/kg ketamine (KM) or 15 mg/kg S-ketamine (SKM). After 60 min, medetomidine was reversed using 0.5mg/kg atipamezole IM. Muscle relaxation, ease of handling, sensitivity to a painful stimulus, palpebral and corneal reflexes, heart rate (HR), and respiratory rate (RR) were measured every 15 min during anesthesia. Time of initial effect, maximal anesthetic effect, first movements, complete wakefulness, first feeding after anesthesia, and quality of induction and recovery periods were compared between protocols. Quality of induction and recovery were excellent with both protocols. HR decreased within the first 15 min but began to rise at 60 min. Marked bradypnea occurred at the beginning of anesthesia but gradually disappeared over time. Muscle relaxation and handling scores increased during anesthesia, whereas sensitivity scores remained 0, regardless of protocol. Pairwise comparisons showed no significant differences between KM and SKM, with the exception of the palpebral reflex at 30 min (P > 0.046). Anesthetic quality was similar using KM or SKM. Both protocols were safe and effective in Testudo spp., but analgesic effects were considered weak. [ABSTRACT FROM AUTHOR]

Abstract It is the first time that Crimean-Congo haemorrhagic fever virus (CCHFV), causing potentially lethal disease of humans, has been reported from the Middle East region and from the tortoise tick Hyalomma aegyptium from a tortoise host, whose epidemiological significance may have remained almost completely overlooked so far. We used RT-PCR to screen for 245 ticks collected from 38 Testudo graeca tortoise individuals. Results of our genetic screening provide unambiguous evidence of occurrence of CCHFV in this region and host, suggesting a potentially important role of H. aegyptium in CCHF epidemiology.