Testudo Hermanni Research Articles

Morphological characteristics of the Hermann's tortoise (Testudo hermanni Gmelin, 1789) in the south-eastern part of Kosovo and Metohija

In this paper we present the results of the analysis of morphological characteristics of the Hermann's tortoise from three localities in south­eastern part of Kosovo and Metohija. 75 adult Hermann's tortoises (33 males and 42 females) have been analyzed. Morphometric analysis has been done on 7 traits. We considered the plastron black pigmentation patterns and frequencies for each pigmentation states were calculated. Females were larger than males for all morphometric traits. Based on the most important dimensions of carapace and plastron length and width, the Hermann's tortoises from the south-eastern part of Kosovo and Metohija are the most similar to the individuals of the population from the southern part of the Balkan Peninsula (the southern part of Serbia and the northern part of Greece). In relation to the populations from central and eastern Serbia, individuals from our studied area have higher values.

Analysis of movement patterns and macrohabitat use in Hermann’s tortoises (Testudo hermanni hermanni, Gmelin 1789) reintroduced in a coastal area dominated by pinewood in eastern Spain

The Hermann’s tortoise is a strictly European species found mostly in areas with Mediterranean and sub-Mediterranean climes and, especially the subspecies T. hermanni hermanni, has to deal with various threats which reduced its geographic distribution to only a few isolated populations. Thus, with the objective of recovering natural populations of the species, numerous reintroduction projects are being carried out throughout its historical and prehistorical geographic distribution. The aim of our study is to evaluate the success of the T. hermanni reintroduction project carried out in the Albufera de Valencia Natural Park since 2011, through the analysis of movements, home range sizes and habitat use in a new environment. The recorded activity pattern presents two peaks, one in spring and another in August, and the hibernation period (from December to February) is the same as observed in other populations. We did not find significant differences in home range sizes between sexes, but a tight relation between home range sizes and body sizes with larger individuals having larger home ranges. This relation was not observed for core areas (Kernel at 50%). Considering the habitat use, we did not find differences between the groups of individuals we defined, but different types of habitats were used differently. “Scrub” was the most frequented (∼50%), followed by “dunes” (∼20%), “mallaes” (∼20%) and “forest” (<20%). Our results can be used to improve the success of future reintroduction projects by directing ideal habitat, timing and carapace size of released Hermann’s tortoises.

Response to Perceptual Novelty in Tortoises-A Preliminary Study

<p class="jbls"><span lang="EN-GB">The ways of responding to novelty have not yet been thoroughly and comprehensively researched in reptiles, and studies undertaken so far have been conducted mainly on laboratory rats. The purpose of this paper is to present results of research carried out on the land tortoises <em>Testudo hermanni</em> and <em>Agrionemys horsfieldii</em>. Current research on tortoises as study subjects indicates that while undertaking environmental exploration these animals rely, to a large extent, on their sight, which is why this study employed visual stimuli to measure the tortoises’ response to novelty. In the course of the experiment, three objects were presented: the first imitated a biologically significant stimulus, the second was biologically indifferent, while the third was variable, and therefore perceived by the animals as novel. The results obtained suggest that the biologically significant and the variable objects elicited a response which involved the animals’ looking more intently at those objects than at the biologically indifferent object. It may be concluded, therefore, that tortoises have demonstrated exploratory responses which varied according to the object presented. While this study was only preliminary, the results obtained substantiate the view that tortoises may be interesting study subjects in comparative psychology.<strong></strong></span></p>

The Genome of a Tortoise Herpesvirus (Testudinid Herpesvirus 3) Has a Novel Structure and Contains a Large Region That Is Not Required for Replication In Vitro or Virulence In Vivo

Testudinid herpesvirus 3 (TeHV-3) is the causative agent of a lethal disease affecting several tortoise species. The threat that this virus poses to endangered animals is focusing efforts on characterizing its properties, in order to enable the development of prophylactic methods. We have sequenced the genomes of the two most studied TeHV-3 strains (1976 and 4295). TeHV-3 strain 1976 has a novel genome structure and is most closely related to a turtle herpesvirus, thus supporting its classification into genus Scutavirus, subfamily Alphaherpesvirinae, family Herpesviridae. The sequence of strain 1976 also revealed viral counterparts of cellular interleukin-10 and semaphorin, which have not been described previously in members of subfamily Alphaherpesvirinae. TeHV-3 strain 4295 is a mixture of three forms (m1, m2, and M), in which, in comparison to strain 1976, the genomes exhibit large, partially overlapping deletions of 12.5 to 22.4 kb. Viral subclones representing these forms were isolated by limiting dilution assays, and each replicated in cell culture comparably to strain 1976. With the goal of testing the potential of the three forms as attenuated vaccine candidates, strain 4295 was inoculated intranasally into Hermann's tortoises (Testudo hermanni). All inoculated subjects died, and PCR analyses demonstrated the ability of the m2 and M forms to spread and invade the brain. In contrast, the m1 form was detected in none of the organs tested, suggesting its potential as the basis of an attenuated vaccine candidate. Our findings represent a major step toward characterizing TeHV-3 and developing prophylactic methods against it. Testudinid herpesvirus 3 (TeHV-3) causes a lethal disease in tortoises, several species of which are endangered. We have characterized the viral genome and used this information to take steps toward developing an attenuated vaccine. We have sequenced the genomes of two strains (1976 and 4295), compared their growth in vitro, and investigated the pathogenesis of strain 4295, which consists of three deletion mutants. The major findings are that (i) TeHV-3 has a novel genome structure, (ii) its closest relative is a turtle herpesvirus, (iii) it contains interleukin-10 and semaphorin genes (the first time these have been reported in an alphaherpesvirus), (iv) a sizeable region of the genome is not required for viral replication in vitro or virulence in vivo, and (v) one of the components of strain 4295, which has a deletion of 22.4 kb, exhibits properties indicating that it may serve as the starting point for an attenuated vaccine.

The tortoises from the Lower Pleistocene palaeontological site of Quibas (Región de Murcia, Spain)

Turtle remains from the Lower Pleistocene palaeontological site of Quibas (Sierra de Quibas, Región de Murcia, Spain) are abundant, but they have not been analyzed taxonomically. Here we determine that all diagnostic specimens belong to Hermann's Tortoise (Chersine hermanni or Testudo hermanni) and they represent the oldest Iberian known occurrence of this taxon.

Contrast echocardiography for analysis of heart anatomy in tortoises

The cardiac anatomy in tortoises depicted on B-mode and color-Doppler should be better differentiated by additional contrast-echocardiography. An intravenous contrast agent (INN-sulphur hexafluoride [SonoVue®]) was injected in four tortoises (three Testudo hermanni, one Agrionemys horsfieldii), via the coccygeal vein, with parallel B-mode echocardiographic examination. The results of the contrast-enhanced echocardiography were compared with the contrast-free B-mode recordings and color-Doppler ultrasound. The use of SonoVue® enabled clearer distinction of the cardiac structures, differentiation of the arising major arteries, identification of wash-out-shunts, and visualization of blood flow patterns throughout the tortoise heart. This study enables the accurate depiction and differentiation of cardiac anatomy in tortoises through the use of a combination of B-mode, color-Doppler and contrast-echocardiography. Basic knowledge of blood flow in the reptile heart is essential to understand echocardiographic anatomy. Blood-flow-patterns and anatomy of the tortoise heart as found in this study will help to establish a basis for further echocardiographic examinations of these animals.

Hematological values for adult eastern Hermann's tortoise (Testudo hermanni boettgeri) in semi-natural conditions.

Hermann's tortoise (Testudo hermanni) is considered near threatened in the wild but, by contrast, it is one of the most popular pet tortoises in Europe. Scant data is reported in the veterinary literature on hematological values for T. hermanni and, to our knowledge, none focused on the subspecies boettgeri (eastern Hermann's tortoise). Published reports are based on small sample populations, and confusion arises when comparing the sampling sites, the anticoagulants, and the counting methods used. The purpose of the current study was to establish the normal mean values and reference intervals for the main hematological parameters for captive adult T. hermanni boettgeri and to evaluate the reliability of a semiautomated blood analyzer for red blood cell count and hematocrit determination. Blood values were determined in 23 adult tortoises using a Neubauer chamber with Natt and Herrick solution; red blood cells and hematocrit were also measured using a semiautomated blood analyzer. Statistical analysis included descriptive statistics, differences between sexes, and agreement between the counting methods. Reference intervals were calculated with the robust method. Wilcoxon signed rank test with continuity correction was used to investigate differences between sexes, and Bland-Altman analysis was performed to compare manual versus semiautomated values. Red blood cells, hematocrit, and hemoglobin determinations were significantly higher in males than in females. White blood cell counts did not show any sex variability. The agreement of manual versus semiautomated determination was considered acceptable for clinical use.

Reference values for hematology and plasma biochemistry variables, and protein electrophoresis of healthy Hermann's tortoises (Testudo hermannissp.)

Hermann's tortoise, Testudo hermanni, is currently on the International Union for Conservation of Nature (IUCN) red list of endangered species. Reptile medicine relies also on laboratory analyses to evaluate health status, but reference ranges for hematology and biochemistry variables and protein electrophoresis in plasma of healthy tortoises are not available. The purposes of this study were to establish reference ranges for select hematologic and biochemical variables in clinically healthy Hermann's tortoises, and evaluate the impact of sex and season. Blood samples were collected from 34 healthy tortoises at the end of September and beginning of July. Blood smears, HCT, concentrations of HGB and select plasma biochemical analytes, select enzyme activities, and plasma protein fractions were evaluated. Reference ranges were determined and checked for influence of sex and sampling time point. Typical reptilian RBC and WBC were observed in blood smears. HCT and concentrations of HGB, uric acid and urea, and ALT and AST activities were significantly higher in males than in females. Concentrations of glucose, uric acid, and phosphate, and AST activity were significantly higher at the beginning of July, whereas concentrations of urea and Cl were higher at the end of September prior to hibernation. The electrophoretic protein fractions included albumin, and α, β, and γ globulins. The reference ranges defined in the present study are useful for clinical tortoise medicine and conservation. Sex and seasonal sampling were identified as factors significantly affecting hematology and blood chemistry analytes; they should be taken into consideration when assessing tortoise health status.

Sighting frequency decreases over two decades in three populations of Testudo hermanni from central Italy

Sighting frequency of tortoises (Testudo hermanni), standardized by the number of person-hours of field effort, was studied between 1992 and 2013 at three different study areas in central Italy. Although the frequency of tortoise sightings fluctuated substantially across years and among study areas, there was a significant declining trend in all the three areas, with GLM analyses showing also a significant interaction between study area and sampling year. The decreasing trend was higher in Castel Fusano than in the other two sites, with yearly frequency of sightings being independent of search effort in all of the study areas. These trends may indicate population declines, because no other explanations seem plausible. Total rainfall from previous autumn to the study period was positively correlated to tortoise sightings in only one locality. The yearly frequency of sightings of juvenile tortoises did not vary significantly among study areas and across years, thus indicating that simple differences in detectability cannot explain the observed patterns. Summer fires and forest overgrowing seem to explain the tortoise decline in two study areas (Castel Fusano and Oriolo) whereas we were not able to identify any specific reason for tortoise decline in the third study area (Manziana).

Cloacal swab sampling is a reliable and harmless source of DNA for population and forensic genetics in tortoises

Testudo graeca, Testudo hermanni and Testudo marginata are endangered species listed in the IUCN Red List and Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Correct identification of species and distinct population units can support sound conservation projects and forensic applications. However, the collection of biological samples is not always easy or harmless. Blood sampling is usually invasive and risky. Salivary swab sampling is commonly used in other reptiles but can be unsafe for tortoises by the reason of their head retraction escape response. Cloacal swab sampling should be easier and less risky. In this study, we genotyped 37 tortoises at eight microsatellite loci and compared the reliability of individual genotypes obtained from blood, cloacal and buccal swabs. Results showed that performances of cloacal samples are comparable with those of buccal and blood samples, and proved that cloacal sampling is an alternative and reliable source of DNA for tortoise genotyping.

Explorations in anatomy: the remains from Royal London Hospital

This paper considers the faunal remains from recent excavations at the Royal London Hospital. The remains date to the beginning of the 19th century and offer an insight into the life of the hospital's patients and practices of the attached medical school. Many of the animal remains consist of partially dissected skeletons, including the unique finds of Hermann's tortoise (Testudo hermanni) and Cercopithecus monkey. The hospital diet and developments in comparative anatomy are discussed by integrating the results with documentary research. They show that zooarchaeological study of later post-medieval material can significantly enhance our understanding of the exploitation of animals in this period

Do Handling and Transport Stress Influence Adrenocortical Response in the Tortoises (Testudo hermanni)?

The goal of this study was to analyze circulating cortisol levels from tortoises (Testudo hermanni) to establish reference intervals and to develop guidelines for the interpretation of the effect of handling and transport stress. Blood samples were obtained from the caudal venous from 23 healthy juvenile tortoises (9 males and 14 females), aged 8–20 years, in basal condition, four weeks prior to and four weeks following handling and short transportation. The study was carried out on the experimental group: 10 tortoises, 4 males and 6 females, and on a control group: 13 tortoises, 5 males and 8 females. Compared to basal values, circulating cortisol concentrations was higher after handling and transport (+286%; P < 0.001), with an increase of +246% (P < 0.001) in males, +236% (P < 0.005) in females, +370% (P < 0.005) in subjects aged 8–12 years, and +240% (P < 0.001) in subjects aged 13–20 years. These observations support the hypotheses that cortisol may act to mediate the effects of handling and transport stress in this species and that four weeks following handling and transport were insufficient to restore their homeostasis.

Endoscopic assisted orchiectomy in Herman's tortoises (Testudo hermanni sp.).

Male tortoises in captivity are often aggressive against other males or females, in particular during mating season related to hormonal influences (testosterone). Castration in males is the treatment of choice in many vertebrate species. A novel technique of minimal invasive castration is presented for Herman's tortoises (Testudo hermanni). The procedure was performed in ten mature males. An endoscope (2.7 mm/30° angled) was inserted into a prefemoral incision on each side. The testicle was resected after ligation of the gubernaculum testis and the spermatic cord using hemoclips. Each testicle could be removed in approximately 20 minutes. Complications like hemorrhage or damage of adjacent tissue did not occur and all animals recovered uneventfully. Using the bilateral prefemoral entrance offers an alternative option for orchiectomy in Herman's tortoises without dissection of the shell. Thus complications like hemorrhage or impaired wound healing followed by sequestration of the bone flap are prevented. Because of the anatomical settings and the risk of tissue damage and time consumption, a unilateral approach is not recommended. Fasting the animals is necessary due to the voluminous gastrointestinal tract of this herbivorous tortoise and emptying the urinary bladder provides more space for manipulations in the coelomic cavity and prevents clipping and cutting of adjacent organs. The magnification via the endoscope is beneficial for orientation in the coelomic cavity, which is formed by the extremely convex carapace. Without this equipment it is challenging to visualize the dorsocaudally located gonads through the relatively small incision and the inserted instruments might block the view at the surgical field. Administration of hemoclips achieved a good hemostasis and the testicles could be resected without major blood loss. The described technique is a gentle method for resection of the testicles in this species and can be adapted to other European tortoise species of equal size.

Déclin d’une population de Tortue d’Hermann (Testudo hermanni hermanni Gmelin, 1789) en milieu forestier. Effets combinés de plusieurs facteurs anthropiques

Decline of a Hermann’s Tortoise (Testudo hermanni hermanni Gmelin, 1789) population in forest. Combined effects of several anthropogenic factors. This study investigates the evolution of a population of Hermann’s tortoises, located on the edge of the massif des Maures, over a period of 30 years through capture– recapture monitoring. It shows a considerable decline of 73 % in the number of adult tortoises between the early 1980s and 1999 ; then from 1999, the population stabilized at around 100 adult tortoises. The fall in numbers during the first period seems to be linked to several factors. The 22 % increase in forest cover between 1983 and 1995 increased the concentrations of eggs laid in some open areas, making it easier for the species’ natural predators to find them. Mechanized clearance of undergrowth carried out in 1989 and 1994 has undoubtedly reduced the size of the population. The presence of dogs has posed an additional threat (97 % of the adult tortoises have bite marks and 7 % are missing limbs). In addition, the impact of dog bites on the tortoises’ shells has, over time, removed individual markings, thereby distorting survival estimates. The stabilization of numbers over the last decade seems to be due to the discontinuation of certain practices (notably mechanized undergrowth clearance). However, the demographic structure (with almost no juveniles) could soon lead to another decrease in population. This situation seems to be specific to this particular population and cannot be applied generally to all the populations in Provence.

Gastrointestinal parasitic burdens in UK tortoises: a survey of tortoise owners and potential risk factors

Despite gastrointestinal parasites being commonly diagnosed in captive tortoises throughout the UK, there is a lack of data regarding the prevalence. The aims of this study were to investigate the...

Genetic Variation and Population Structure in the Endangered Hermann's Tortoise: The Roles of Geography and Human-Mediated Processes

The Hermann's tortoise (Testudo hermanni) is an endangered land tortoise distributed in disjoint populations across Mediterranean Europe. We investigated its genetic variation by typing 1 mitochondrial locus and 9 nuclear microsatellites in approximately 300 individuals from 22 localities. Our goal was to understand the relative impact of natural and human-mediated processes in shaping the genetic structure and to identify the genetic priorities for the conservation of this species. We found that 1) all geographic areas are highly differentiated, mainly as a function of their distance but with a clear genetic discontinuity (F st values larger than 0.4) between the Eastern and the Western subspecies; 2) the contact zone between subspecies is located farthest to the west than previously believed, and it probably coincides with the delta of the largest Italian river; 3) extinction events due to climatic conditions in the Upper Palaeolithic and subsequent human-mediated translocations in the Neolithic possibly explain the unexpected similarity among Spain, Sicily, and Corsica. For conservation purposes, the large majority of genetic pools appears native although hybridization among subspecies, related to extensive 20th century trade of tortoises across Europe, is observed in Spain and some Italian samples. Most populations do not seem at immediate risk of low genetic variation, except the French population, which has very low nuclear genetic diversity (heterozygosity = 0.25) and where 50 out of 51 sampled animals shared the same mitochondrial sequence. In general, restocking and reintroduction plans should carefully consider the genetic background of the individuals.

Taxonomic and functional response of a Mediterranean reptile assemblage to a repeated fire regime

Wildfires are common disturbances that have a major impact on ecosystems. Recent decades have seen an increase in fire frequency and extension due to the combined effects of climate change and land-use history. We studied the taxonomic and functional response of a reptile assemblage to repeated fires in southern France to understand shifts in dominant species and diversity, as well as the mechanisms that underlie responses according to functional traits of species. In the spring of 2010, we sampled reptiles in areas with three types of fire regime: unburned, burned once (2003) and burned 4–5 times (last fire in 2003) along a fire history of 51-years period. With this field sampling design, we examined the intermediate disturbance hypothesis and the habitat accommodation model of succession as methods to predict reptile responses to natural fire regimes. We also compared habitat structure at the study area between 1944 and 2006 to certify that repeated-fire regimes have modified the habitat for reptiles. The comparison of the habitat structure between both periods demonstrated that repeated-fire regimes modified the landscape from a homogeneous sparse forest to a contrasted heterogeneous mixture of scrubland and dense forest. We found a loss of reptile diversity after one and multiple fires, a result that contradicts the intermediate disturbance hypothesis. Reptile composition differed among the three fire regimes: there was a shift in dominant species and a reduction of beta diversity related to an increase in the number of fires. We also observed a functional response to repeated fires, with an increased frequency of insectivorous reptiles, which live in open areas, are specialists in their ecological niche, and have a short lifespan. These results suggest that reptile replacement according to fire regime accounts for a habitat accommodation model following particular traits of species. Our study indicated that areas subjected to repeated fires have a more strictly Mediterranean reptile assemblage than unburned areas, due to the ability of Mediterranean species to survive thermal environments in open (burned) areas. At a regional scale, changes in dominant species between unburned and repeatedly burned areas might be an argument for maintaining a patchwork of areas burned at variable intervals. However, the increase in fire frequency and extension suggests a future scenario of extinction for species negatively impacted by fire, such as the endangered Hermann’s tortoise Testudo hermanni, for which the study area is home to one of the last native populations in the western Mediterranean.

Cystoscopic sex identification of posthatchling chelonians

To describe a noninvasive technique for sex identification of posthatchling chelonians and to assess its safety in Hermann's tortoises (Testudo hermanni). Validation study and clinical trial. 15 recently dead posthachling chelonians and 25 healthy posthatchling Hermann's tortoises. Cystoscopy was performed on both dead and anesthetized live chelonians. Dead chelonians ranged in body weight from 32.4 to 75.1 g (0.07 to 0.17 lb; median, 45.7 g [0.10 lb]). Dead chelonians were dissected immediately after cystoscopy, and gonads were collected for histologic examination. Urinary bladder was macroscopically evaluated in situ to assess its integrity after retrograde injection of saline (0.9% NaCl) solution. Hermann's tortoises ranged in body weight from 27.3 to 57.8 g (0.06 to 0.13 lb; median, 37.0 g [0.08 lb]). Cystoscopic examination of live tortoises was performed following induction of general anesthesia with a mixture of morphine, dexmedetomidine, and ketamine administered IM. The Pearson coefficient was used to assess the consistency between procedure time and body weight; κ statistic was used to evaluate agreement between sex identified by cystoscopy and histologic examination beyond that expected by mere chance. Visualization of gonads was feasible through the thin, transparent urinary bladder wall in all the animals evaluated in this study. Blinded histologic examination confirmed the results of cystoscopic gonad identification in all dead chelonians (κ = 1.0). The urinary bladder did not have evidence of macroscopic leakage or microscopic alterations of normal tissue architecture within the representative sections chosen for histologic examination. In live tortoises, median procedure time (range) was 90 (39 to 345) seconds. No significant correlations were found between procedure time and body weight. Cystoscopy performed by means of rigid endoscopy with fluid instillation was found to be an effective method for sex identification of immature chelonians. Furthermore, no complications were observed when this procedure was performed in vivo.

Assessment of dual-energy x-ray absorptiometry for use in evaluating the effects of dietary and environmental management on Hermann's tortoises (Testudo hermanni)

To assess dual-energy x-ray absorptiometry (DXA) for evaluating effects of diet and environment on bone mineral density in Hermann's tortoises (Testudo hermanni). 26 Hermann's tortoises within 1 month after hatching. Group 1 was housed in an artificial setting and fed naturally growing vegetation. Group 2 was housed in an artificial setting and fed vegetables grown for human consumption. Group 3 was maintained in an outside enclosure and fed naturally growing vegetation. After 10 months, pyramidal growth, body weight, and adverse conditions were assessed. Bone mineral density (BMD) of the axial and appendicular skeleton, shell, vertebral column, and pelvis was measured via DXA. Group 2 had the highest mean ± SD body weight (65.42 ± 30.85 g), followed by group 1 (51.08 ± 22.92 g) and group 3 (35.74 ± 7.13 g). Mean BMD of the shell varied significantly among groups (group 1, 0.05 ± 0.03 g/cm(2)•m; group 2, 0.09 ± 0.15 g/cm(2)•m; and group 3, undetectable). The BMD of the axial and appendicular skeleton, vertebral column, and pelvis did not differ significantly among groups. Pyramidal growth was highest in group 1 and not evident in group 3. Tortoises raised in artificial conditions did not have deficits in BMD, compared with results for outdoor-housed hibernating tortoises. Supplemental calcium was apparently not necessary when an adequate photothermal habitat and plant-based diet were provided. Higher BMD of captive-raised tortoises was morphologically associated with a higher incidence of pyramidal growth in captive-raised groups.

Characterization and quantification of the influence of season and gender on plasma chemistries of Hermann’s tortoises (Testudo hermanni, Gmelin 1789)

The present study is the first to monitor plasma chemistries of a group of 30 Hermann’s tortoises (HTs) over two summer seasons to characterize and quantify seasonal and gender-related influences. The following analytes were analyzed: ALT, ALP, AST, BA, CA, CHO, CK, LDH, GLU, GLDH, P, TP, TRIG, urea and uric acid. Two-way repeated-measures ANOVA, mean values, confidence intervals were calculated. The overall results showed distinct patterns and somewhat unexpected results concerning the plasma chemistry changes induced by season and gender in a majority of routine biochemical analytes. TRIG, CHO, CA, and P were significantly higher in females. AST, ALT, BA, LDH and GLDH showed an increase in males in midsummer. ALP showed a significant and analogous seasonal variation for both sexes. GLU showed a moderate increase during midsummer. Uric acid and urea concentrations showed a distinct seasonal variation. For some analytes, fitted curves could be calculated describing the seasonal variation.