Ultrasonographic diagnosis of pericardial effusion and atrial dilation in a spur-thighed tortoise (Testudo graeca)

Ultrasonography was used to diagnose pericardial effusion, atrial dilatation and liver masses in a spur-thighed tortoise which was more than 80 years old and suffering from posthibernation anorexia, lethargy, oedema and pneumonia. The tortoise was treated twice with frusemide and ceftazidime for the pneumonia, resulting each time in a temporary remission for about a month. After a further recurrence, the animal was euthanased and the lesions predicted by ultrasound were confirmed postmortem. It is suggested that ultrasound may be useful for the differentiation of cardiac problems from other causes of posthibernation lethargy in the tortoise.     

Diagnosis and surgical repair of stifle luxation in a spur-thighed tortoise (Testudo graeca).

This report describes stifle luxation resultant from multiple ligamentous injuries in a spur-thighed tortoise (Testudo graeca). Despite an absence of obvious trauma in the case history, the tortoise presented with acute, unilateral hind limb lameness. Diagnosis of stifle luxation was based on cranial drawer motion, increased rotational joint instability. and radiographic findings. Joint stabilization was accomplished by a combination of cranial cruciate reconstruction using an autograft of lateral vastus muscle in an over-the-top procedure and lateral imbrication of the joint capsule. Postoperative management included restriction of limb movement for 6 wk and parenteral chondroprotective regime. Treatment was effective in restoring clinically normal function to the limb.     

Seasonal variation in blood biochemistry of long-term captive Mediterranean tortoises (Testudo graeca and T hermanni)

Reference values for five blood chemistry parameters in 18 Mediterranean tortoises of two species, Testudo graeca and T hermanni, were determined on up to 10 occasions during the year. Statistically significant seasonal variations were demonstrated in blood urea and blood glucose. Seasonal variations were demonstrated in blood urea and blood glucose. Seasonal variations in total plasma proteins, lipids and cholesterol, however, were limited to gravid females. The study also suggested that three energy sources were available to the tortoise during hibernation, lipids stored in the fat body, endogenous protein degradation and glucose derived from hepatic glycogen.     

Atypical systemic mycobacteriosis in Hermann's tortoises (Testudo hermanni)

Background: Hermann's tortoise (Testudo hermanni) is a near threatened species found throughout southern Europe. Captive breeding of endangered species is subject to health risks, such as the stress of captivity and ambient conditions that may favor the emergence of infectious diseases. An episode of systemic atypical mycobacteriosis was investigated in six Hermann's tortoises from a captive population. The symptoms appeared in the form of anorexia, weakness, and lethargy, with inflammatory and edematous lesions of the hind limbs and tail. Four of the affected tortoises were euthanized and necropsied. Methods: Blood samples and joint aspirates were obtained for assessment. Euthanasia of four affected animals was performed. Postmortem examination included necropsy, histopathology, and mycobacterium detection by culture and polymerase chain reaction (PCR). In the other two animals, only hematology and blood biochemistry was carried out. Results: Hematological and blood biochemistry analyses gave inconsistent results. In joint aspirates, mononuclear cells with phagocytozed bacillary structures were observed. Ziehl-Neelsen stain revealed the presence of acid-fast bacteria. Granulomatous lesions were observed in liver, lungs, spleen, heart, muscle, kidney, and ovaries. Culture and identification were positive for mycobacteria and molecular sequencing led to the identification of Mycobacterium nonchromogenicum. Conclusions and clinical relevance: Although mycobacteriosis is rare in reptile collections, strict hygienic and prophylactic measures in species of high ecological value must be performed, especially if the animals are to be used for reintroduction projects.     

Pulmonary candidiasis caused by Candida albicans in a Greek tortoise (Testudo graeca) and treatment with intrapulmonary amphotericin B.

An adult female Greek tortoise (Testudo graeca) presented with dyspnea, lethargy, and anorexia. Severe unilateral pulmonary candidiasis was diagnosed and confirmed by histologic and microbiologic evaluations. Initial treatment with ketoconazole resulted in plasma elevations of aspartate aminotransferase, lactate dehydrogenase, and bile acids consistent with imidazole-induced hepatotoxicity. Plasma chemistry abnormalities resolved upon withdrawal of the drug. Temporary osteotomy permitted access to the diseased lung and facilitated intrapulmonary catheterization. Intrapulmonary amphotericin B therapy at 0.1 mg/kg s.i.d. for 34 days proved to be both safe and effective in this case.     

Differences in helminth infections between captive and wild spur-thighed tortoises Testudo graeca in southern Spain: a potential risk of reintroductions of this species

Although the spur-thighed tortoise, Testudo graeca, is one of the most widely distributed species of tortoises, its natural populations are threatened through its whole range. Particularly at south-eastern Spain, the species is mainly threatened by habitat destruction and over-collection, given that this chelonian has been traditionally considered an appreciate pet. As south-eastern Spanish wildlife recovery centers shelter hundreds of captive animals mainly coming from illegal trade or captive-bred, there is a strong debate about what to do with these animals: maintaining them in captivity all along their lives or reintroducing them to wildlife. It is well known that the reintroduction of captive animals supposes a risk for the wild population due to the uncertainty of their genetic origin and to the possible spread of infectious diseases. However, despite the increasing evidence that infectious agents are a potential health hazard for wildlife, little is known about the risk that introduced parasites could suppose for the wild populations of spur-thighed tortoise. The present study investigates for the first time the presence of helminth eggs and worms in faeces from 107 wild and captive individuals collected from mid-March to mid-June 2010, and relates the findings to different environmental and host variables. Sixteen oxyurid species and the ascarid Angusticaecum holopterum were identified. This last nematode and the oxyurid species Tachygonetria palearticus and T. seurati had not been reported in Spanish wild T. graeca previously. The prevalence of oxyurid eggs and worms were 94% and 70%, respectively; while, ascarid eggs and worms were found in 26% and 5% of tortoises, respectively. Ascarid infections affected mostly captive animals and were associated to caparace deformities and symptoms of upper respiratory tract disease (p<0.05). Oxyurid infections were not associated to negative health traits and prevalence increased with age. In free-living tortoises, the distribution of pharingodonid genera also varied according to habitat; moreover, T. longicollis, T. pusilla, T. conica, T. robusta and Mehdiella stylosa where significantly more frequent in wild compared to captive tortoises (p<0.05). Study results highlight important differences in the nematode fauna of captive and free-living tortoises and questions one more time if the reintroductions of captive animals suppose a risk for the wild population since the former ones can harbor and distribute among free populations pathogens like ascarid nematodes.     

Scanning electron microscopic and histological features of the oral cavity of the Egyptian tortoise (Testudo kleinmanni)

This investigation was led to depict the structural and functional adaptations of the oral cavity of herbivorous Egyptian tortoises using scanning electron and light microscopes. The SEM showed that the triangular papillary tongue possessed three conical papillary subtypes: the rectangular conical on the tip, the round conical on the rest of the dorsal lingual surface and the elongated conical on the caudal portion of the lingual wing. The presence of the serrated lips with their valves compensated for the absence of the teeth. The rostral part had a vomeronasal opening while the middle part had the choana, but the caudal part had numerous openings of the salivary glands. There are three palatine folds: a single median palatine fold, two peripheral palatine folds and the choanal fold. The current histological results show the keratinized dorsal lingual surface, in which the keratinized layer extended to cover the papillae. Two types of lingual glands, according to their position, are papillary superficial and deep lingual glands. Papillary or superficial glands open in the interpapillary spaces via narrow openings, while the deep glands are surrounded by well-developed muscles and open via wide openings on the dorsal lingual surface. An entoglossal cartilaginous structure of hyaline cartilage was found in the mid- and hindtongue, with numerous chondrocytes lodged within the lacunae. Our results conclude that the oral cavity of the herbivorous Egyptian tortoise was adapted to the dietary and vigorous demands of the Egyptian fauna.     

Feasibility and variability of corneal esthesiometry in Hermann's tortoises (Testudo hermanni) depending on age and body weight

BackgroundHermann's tortoise is one of the most popular reptiles kept as pet which underlines the importance to reinforce the information needed to provide advanced and adequate veterinary care in exotic animal species. Therefore, the purpose of this study, performed in Testudo hermanni, was to evaluate corneal touch threshold (CTT) and its feasibility according to age and body weight.MethodsFifty-one healthy tortoises were classified in 2 groups (≤2 years [young; n = 25] and 8–10 years [subadult; n = 26]). Central CTT was measured by means of a Cochet-Bonnet esthesiometer and defined as the filament length required to elicit a blink in at least 3/5 applications. CTT feasibility according to body weight was also evaluated by diving the individuals in weight groups consisting of “lighter” (<400 g [n = 31]) and “heavier” (≥400 g [n = 20]) animals.ResultsMean CTT was 5.99 cm for the whole population (90% CI: 5.87–6.11), being 5.98 cm for young (90% CI: 5.81–6.15) and 6 cm for all subadult (90% CI: 6.00–6.00) tortoises, and 5.98 cm for lighter (90% CI: 5.84–6.13) and 6 cm for all heavier (90% CI: 6.00–6.00) tortoises. No statistical differences were detected between age and weight groups (P = 0.159 and P = 0.159, respectively). Three animals presented unilateral faint fluorescein uptake postesthesiometry (3/51; 2.9%) that resolved spontaneously within 48 hours.Conclusions and clinical relevanceCochet-Bonnet esthesiometer was a safe means of confirming high corneal sensitivity in all tortoises, which was high regardless of age. Increasing filament lengths would ultimately be required to determine the true corneal sensitivity scope of Testudo hermanni.     

Development of species-specific PCR techniques for the detection of tortoise herpesvirus.

Previously, a nested polymerase chain reaction (PCR) was employed with consensus degenerate primers targeting highly conserved motifs within herpesviral DNA polymerase genes to detect a newly described tortoise herpesvirus. However, nucleotide sequence information obtained from the final amplified fragment was restricted to a small region of 181 bp. In the present study, additional sequences flanking this segment were determined from a PCR product successfully amplified using a set of known degenerate primers, which covered a 692-bp region within the tortoise herpesviral DNA polymerase gene. Polymerase chain reaction primers for specific amplification of the tortoise herpesviral DNA were designed on the basis of these nucleotide sequences and successfully amplified tortoise herpesviral DNA from the tissues of tortoises that were well characterized histopathologically with herpesviral infection. The lower limit of detection was 1,000 herpesviral DNA equivalents in the presence of normal tortoise genomic DNA. Furthermore, a more sensitive and specific PCR technique for the identification of herpesviral infections in tortoises was developed employing a heminested form, which will enable the detection of latent infections or herpesvirus carriers in tortoises.     

ULTRASONOGRAPHY OF THE CALIFORNIA DESERT TORTOISE (Xerobates agassizi): ANATOMY AND APPLICATION

Ultrasonographic examinations were performed on healthy California desert tortoises to evaluate the utility of the anatomic acoustic windows available and to gain information about the normal echoanatomy of the tortoise. A major objective was to evaluate the potential clinical value of this diagnostic tool in these species. The soft tissue areas of the integument were used as acoustic windows. Three imaging approaches were described as mediastinal, axillary, and inguinal openings. The mediastinal and axillary approaches offered good visualization of the heart, liver, and gallbladder. The inguinal window allowed imaging of the intestines, urinary/accessory bladders, kidneys, and gonads. The pancreas and spleen could not be visualized. A tortoise with pericardial effusion and a tortoise with hepatic lipidosis are presented to illustrate clinical applications of ultrasound in diagnosing diseases of the tortoise.     

Molecularbiological diagnosis of herpes virus infection of a juvenile Russian tortoise (Agrionemys horsfieldii) with skin and lung lesions

Herpesvirus infections are significant in the care of turtles and tortoises. Clinical signs range from unspecific symptoms, due to the variety of organ manifestations, to the "classical" picture of rhinitis-stomatitis. The presented case study showed the typical disease only with respect to clinical symptoms following hibernation, but lacks stomatitis, erosions or plaques in the oral mucosa. On the other hand, skin lesions on the extremeties, causative with herpesvirus infection, could be diagnosed. In this case study, various symptoms, sampling procedures and diagnostics using two different PCR methods are presented. Following hibernation, samples from a Russian tortoise (Agrionemys horsfieldii) were taken ante mortem and post mortem and screened with respect to virology, pathology, bacteriology and parasitology. DNA-fragments specific for tortoise herpesvirus were detected in various organs and body liquids. Furthermore basophilic intranuclear inclusion bodies were found. The bacteriological examination showed a high level of Pasteurella testunis in the lungs. By parasitological examination nematodes (Oxyridae) were diagnosed. A potential prophylaxis tortoise herpesvirus is discussed.     

Identification of a novel herpesvirus from a California desert tortoise (Gopherus agassizii)

Herpesviruses are significant pathogens of tortoises, causing upper respiratory tract disease and necrotizing stomatitis, with infections often associated with high mortality rates. Herpesvirus infection in a captive California desert tortoise (Gopherus agassizii) was detected by light microscopic observation of intranuclear inclusion bodies in various tissues followed by transmission electron microscopic observation of herpesvirus-like particles, and amplification of herpesvirus nucleic acid sequences using polymerase chain reaction. Using an indirect enzyme linked immunosorbent assay, anti-tortoise herpesvirus antibodies were detected one month after initial onset of clinical signs. This novel herpesvirus is distinct from the previously described tortoise herpesvirus (tortoise herpesvirus-1, THV-1) sharing 83% sequence identity of 60 amino acids of a portion of the DNA polymerase gene and 79% sequence identity across 120 amino acids of a portion of the ribonucleotide reductase gene. Similar to THV-1, this novel herpesvirus, tortoise herpesvirus-2 (THV-2), also clusters with the alphaherpesviruses.     

Liver Size,Bodyweight, and Tolerance to Acute Complete Occlusion of Congenital Extrahepatic Portosystemic Shunts in Dogs

Objective— To investigate the relationship between preoperative liver size, bodyweight, and tolerance to shunt occlusion in dogs with congenital extrahepatic portosystemic shunt(s) (CPSS). Study Design— Longitudinal cohort study. Animals— Dogs with CPSS (n=35). Methods— Ultrasonography was used to measure preoperative maximum transverse dimension of the liver (TS) of each dog. Intraoperative portal pressures were measured, before and after CPSS occlusion, via a jejunal vein catheter. Tolerance to shunt occlusion was judged on gross visceral observations, and on changes in portal pressure, central venous and mean arterial pressures. Results— TS was significantly related to bodyweight (P<.05). Mean ratios for TS/bodyweight were calculated for dogs tolerant and intolerant of acute complete shunt occlusion. Dogs tolerant to occlusion had significantly higher TS/bodyweight ratios than dogs intolerant to occlusion (P=.025). Dogs with a TS/bodyweight ratio of >7 were more likely to tolerate CPSS occlusion than dogs with a TS/bodyweight ratio of <5 (P=.036). A model was generated to predict portal pressure rise after shunt occlusion, based on liver dimensions and bodyweight (R=0.668). Intestinal oxygenation did not correlate significantly with tolerance to CPSS occlusion (P=.29). Conclusion— In dogs with CPSS, liver size (relative to bodyweight) is significantly greater (P=.025) in dogs that are tolerant of full ligation than intolerant of occlusion. Clinical Relevance— Preoperative measurement of bodyweight and liver size help indicate the likelihood of tolerance to acute complete occlusion of CPSS in dogs.     

Comparison of 11 herpesvirus isolates from tortoises using partial sequences from three conserved genes

Herpesviruses are an important cause of epidemic disease in tortoises. There are at least two serologically distinct herpesviruses capable of infecting tortoises. Methods for the diagnosis of herpesvirus infections in tortoises include virus isolation and a number of different PCRs. We have compared 11 virus isolates collected from various species in different countries over several years using sequences from three different viral genes. During this study we used four different PCR protocols described for the diagnosis of herpesvirus infections in tortoises. The protocols used included two based on portions of the DNA polymerase gene, one targeting the UL5 homologue, and one targeting the UL39 homologue. Comparison of the methods showed that the tortoise herpesvirus-specific protocols were all serotype specific. Sequences of the obtained amplicons were compared with one another and with sequences of herpesviruses available in GenBank. The sequence alignments showed that the tortoise herpesviruses were most closely related to members of the subfamily Alphaherpesvirinae. They also showed that the tortoise isolates could be clearly divided into two genogroups.     

Herpesvirus epidemic in Greek (Testudo hermanni) and Moorish land tortoises (Testudo graeca) in Switzerland

Based on an epidemic in Switzerland, the histopathological findings of a viremic Herpes infection in Spur-tailed and Spur-thighed mediterranean land tortoises are described for the first time. Horizontal viral transmission has been proved; the vertical transmission is suggested. An impression smear of the tongue enables an early clinical diagnosis.     

Patterns in the Distribution of Southern African Terrestrial Tortoises (Cryptodira: Testudinidae)

Distribution maps for the ten southern African species of land tortoise have been prepared using published sources and recent museum records. Three hundred and thirty-six new records have been added to these maps by a recent Cape Province tortoise survey, and a further 65 of the older records have been re-confirmed. A relatively clear picture of the range of each species is now available, although the records in South West Africa are not yet adequate enough to resolve the problems posed by the vague localities published at the beginning of the century. The Cape Province of South Africa clearly stands out as the world's richest area in terrestrial testudinids.     

Evanescent-wave biosensor for field serodiagnosis of tortoise mycoplasmosis

Disease has become an increasingly important issue for wildlife management over the past two decades. Adequate surveillance is fundamental for disease prevention and control, thus there is an increasing need for diagnostic assays for wildlife management. The objective of this study was to evaluate the performance of a field-portable biosensor adapted for rapid detection of specific antibodies in tortoise plasma that reflect a history of exposure to Mycoplasma agassizii, which is an agent of tortoise upper respiratory tract disease. Banked plasma samples were tested in two blinded trials, and the parameters that define the reliability of a diagnostic test were estimated based on externally validated tortoise plasma controls. The mean sensitivity of the biosensor (ability to identify exposed tortoises in the group of all exposed individuals) was 78%; the mean specificity (unexposed individuals with negative test result, out of all unexposed individuals tested) was 73%; the mean positive predictive value (exposed individuals with positive test, out of all individuals with positive test) was 82%; the mean negative predictive value (unexposed individuals with negative test, out of all individuals with negative test) was 68%. In a 15-min field-portable format, the biosensor was able to discriminate between true seropositive (n=34) and true seronegative (n=23) tortoise plasma with overall accuracy of 84%. The goals established for the tortoise population can help managers decide whether potential diagnostic errors should impact management decision-making, and whether the benefits of the field-portable format of the biosensor assay outweigh any potential disadvantages.     

Ultrasonographic measurement of normal renal parameters

Both kidneys were imaged ultrasono-graphically in each of 100 normal adult dogs. Small but significant differences were found in the dimensions of the left and right kidneys and in renal dimensions of male and female dogs of similar bodyweight. There was a statistically significant correlation between renal length and bodyweight (r = 0–84, P<0–001). The relationship between renal length and bodyweight was linear except at very low or very high bodyweights. There was also a statistically significant correlation between renal volume and bodyweight (r = 0–91, P<0–001), and the relationship remained linear even at extremes of bodyweight. Graphs could therefore be constructed for renal lengthy bodyweight and renal volume/bodyweight with 95 per cent confidence limits.