Endoscopic sexing in turtles and tortoises: 467 cases (2007-2017).

BACKGROUND: This multi-institutional retrospective study evaluated the feasibility and safety of endoscopic sex identification in 467 turtles and tortoises, representing 10 species. METHODS: Medical records of turtles and tortoises that underwent endoscopic sex identification at the University of Georgia, New England Aquarium and Turtle Conservancy were reviewed for presurgical management, anaesthesia, endoscopic equipment and surgical techniques, endoscopic results and complications. RESULTS: The majority of animals weighed less than 200 g, were fasted and anaesthetised using an injectable combination of ketamine, dexmedetomidine and morphine or hydromorphone, supplemented by local lidocaine at the prefemoral site. Anaesthetic reversal using atipamezole alone or in combination with naloxone was routine. For uncomplicated procedures, mean total anaesthesia, surgery and recovery times were 22, 4 and 18 minutes, respectively. All animals were placed in lateral recumbency for a prefemoral endoscopic approach to the coelom using a rigid telescope and sterile fluid infusion to visualise the gonads. Sex identification was definitive in 99.4% (n = 464) of the animals. Iatrogenic bladder perforation was the most common complication (n = 5), which necessitated extended anaesthesia and surgical time for repair. Only a single anaesthetic-related death was reported, which was associated with human error and drug overdose. CONCLUSIONS: This is the first large-scale study to retrospectively evaluate endoscopic sex identification in multiple chelonian species. Results suggest that endoscopic sexing is a safe, accurate and practical means for sex identification in turtles and tortoises, and represents a valuable tool in their reproductive management.

Successful outcome of shell fractures in freshwater turtles treated with plate or screws and wire fixation techniques: 51 cases (2014-2019).

OBJECTIVE: To evaluate the outcome of surgical fixation of shell fractures in rehabilitated wild freshwater turtles. ANIMALS: 51 freshwater turtles with 86 shell fractures. PROCEDURES: The medical record database of a wildlife rehabilitation center in Wisconsin was searched from 2014 through 2019 for records of freshwater turtles with shell fractures repaired with a plate technique, screws and wire technique, or both. Signalment, fracture location, therapeutic approach (including the type of hardware used for repair), dry-docking duration, time to hardware removal, postremoval care, and outcome were evaluated. RESULTS: 36 of 51 (71%) turtles with shell fractures experienced successful fracture healing following surgical hardware fixation, and 33 (65%) were released. Shells of 38 (75%) turtles were repaired with plates only, 5 (10%) turtles with wire only, and 8 (16%) turtles with a combination of plates and screws and wires. Of the 51 turtles, 13 (25%) did not survive > 4 weeks following hardware repair, leaving 38 animals available to assess fracture healing. Median time to start staged removal was 42 days (range, 35 to 49 days) and to complete removal of the applied hardware was 56 days (range, 26 to 77 days). Complications associated with placement of the hardware occurred in 6 of 38 (16%) turtles. Complications included screw hole infection (4/38 [11%]), fracture necrosis (1/38 [3%]), and deep screw hole penetration (1/38 [3%]). CLINICAL RELEVANCE: Results indicated that shell fractures in freshwater turtles treated with surgical fixation techniques had a successful outcome. Most complications were minor, and fractures improved with time, resulting in acceptable fracture healing for release.

Circulating miRNome of Trachemys scripta after elective gonadectomy under general anesthesia.

Post-surgical management is an important issue in veterinary medicine, requiring biomarkers with high sensitivity and specificity for timely and effective treatment. Emerging evidence suggests that miRNAs are promising stress- and pain-related markers. The aims were to profile the circulating miRNA signature in plasma of turtles (Trachemys scripta) and point out potential candidate biomarkers to assess the status of the animal. The plasma of female turtles underwent surgical gonadectomy were collected 24 h pre-surgery, and 2.5 h and 36 h post-surgery. The expression of miRNAs was profiled by Next Generation Sequencing and the dysregulated miRNAs were validated using RT-qPCR. The diagnostic value of miRNAs was calculated by ROC curves. The results showed that 14 miRNAs were differentially expressed over time. RT-qPCR validation highlighted that 2-miR-499-3p and miR-203-5p-out of 8 miRNAs tested were effectively modulated. The Area Under the Curve (AUC) of miR-203-5p was fair (AUC 0.7934) in discriminating pre- and 36 h post-surgery samples and poor for other time points; the AUC of miR-499-3p was excellent (AUC 0.944) in discriminating pre-surgery and 2.5 h post-surgery samples, and fair in discriminating pre-surgery and 36 h post-surgery (AUC 0.7292) and 2.5 h and 36 h post-surgery (AUC 0.7569) samples. In conclusion, we demonstrated for the first time that miRNAs profile changes in plasma of turtles underwent surgical oophorectomy and identified miR-203-5p and miR-499-3p as potential candidate biomarkers to assess animals' status. Further studies are necessary to confirm their diagnostic value and to investigate functional and mechanistic networks to improve our understanding of the biological processes.

Tissue tolerance to cyanoacrylate adhesives for closure of surgical wounds in Trachemys scripta skin.

BACKGROUND: The aim of this study was to evaluate the histological tolerance of medical cyanoacrylate adhesives (CAs), butyl-CA and octyl-CA, applied by two different methods in surgical skin incisions of T. scripta. METHODS: A two-trial controlled blind study each one including 12 female adult T. scripta turtles. Two 1-cm long full-thickness surgical skin incisions were made on the dorsal aspect of each hind limb. In trial 1 CAs were applied superficially over the apposed wound edges; each animal's four wounds were closed using butyl-CA, octyl-CA and ethyl-CA or absorbable suture as controls. In trial 2 adhesives were applied holding wound edges everted, and octyl-CA was replaced by a non-sutured control wound. At each time point (2, 7, 14 and 21 days) all wounds from three randomly chosen animals were biopsied. RESULTS: Medical grade CAs did not induce any sign of tissue damage and/or inflammatory reaction attributable to histotoxicity independently of the method of application. Wound dehiscence rate was higher when CAs were applied superficially over the apposed wound edges. CONCLUSIONS: Octyl and butyl-CA are well tolerated by turtles' soft skin but because of their high rate of dehiscence they are not recommended in high tension wounds; use in low tension wounds needs further investigation.

SPINAL ANESTHESIA IN GREEN SEA TURTLES (CHELONIA MYDAS) UNDERGOING SURGICAL REMOVAL OF CUTANEOUS FIBROPAPILLOMAS.

Techniques for anesthesia of green sea turtles (Chelonia mydas) are required for medical treatment. The use of spinal anesthesia has been reported in a few species of turtles for different purposes. The objective of this study was to evaluate the use of 2% lidocaine for spinal anesthesia of green sea turtles undergoing surgical removal of cutaneous fibropapillomas. Ten free-ranging green turtles presenting with cutaneous fibropapillomas were included in the study. Animals were accidentally captured or rescued by local fishermen and brought to the Ubatuba Research Base (Sao Paulo, Brazil) of the Brazilian Sea Turtle Conservation Program for rehabilitation. Animals were administered 2% lidocaine (0.2 ml/10 cm of carapace) in the epidural/subarachnoid space of the tail and monitored throughout surgery. The technique was effective for all animals, with fast onset of motor and sensory blockade (3 ± 1.76 min) and relatively fast recovery time (83.9 ± 16.2 min). Fibropapillomas were removed from all animals with no signs of pain (i.e., no behavioral response during surgical procedure, such as head and forelimb movement, showing discomfort) and they were all rehabilitated and successfully returned to their natural habitat. The technique was considered effective, safe, and affordable for use on green turtles undergoing surgical removal of cutaneous fibropapillomas.

SURGICAL ANATOMY OF CELIOTOMY APPROACHES TO THE STOMACH IN THE COMMON SNAPPING TURTLE (CHELYDRA SERPENTINA).

Entanglement in or ingestion of fishing gear is a common cause of morbidity and mortality in chelonians. Commercial and recreational fishing activities exert bycatch pressures sufficient to cause population declines in the common snapping turtle (Chelydra serpentina) and the alligator snapping turtle (Macrochelys spp.). Gastrotomy for the removal of fish hook foreign bodies from the stomach of freshwater turtles typically requires a plastron osteotomy but in sea turtles has been successfully accomplished via soft tissue approaches. This study compared the anatomy and feasibility of soft tissue surgical approaches to the stomach in the common snapping turtle in both the prefemoral and the axillary regions. Anatomical dissections were performed on cadavers of five adult common snapping turtles, and the surgical anatomy of the left axillary and left prefemoral regions was characterized. The left axillary approach required extensive transection of musculature and provided poor exposure of the coelomic cavity and stomach. In contrast, the left prefemoral approach was simple to perform and provided sufficient access to the stomach for gastrotomy. The prefemoral celiotomy has significant advantages over the axillary celiotomy in the common snapping turtle and should be considered the surgical approach of choice for gastrotomy in this species.

Orchiectomy in Testudo species: technical aspects and effect on courtship behaviour.

Courtship behaviour of tortoises includes biting, ramming, chasing, same-sex sexual behaviour, and it commonly is excessive in European tortoise species kept as pets. As a result, males are kept individually. To assess the effect of orchiectomy on excessive courtship behaviour in European tortoises, 50 tortoises (Testudo graeca , T hermanni and T marginata ) underwent endoscopically assisted orchiectomy from April through September 2013 to 2017. Statistical analyses were performed in relation to species, body mass, testicular size and season and owners were invited to fill out a follow-up questionnaire. Body mass of the tortoises ranged from 334 to 2645 g (mean 1056 g) and the age from 5 to estimated 60 years. Testicular length ranged from 1.4 to 7.0 cm (mean 2.7 cm) and testicular mass from 0.6 to 12.6 g (mean 3.9 g). A complete or partial reduction (allowing group-housing) of the excessive courtship behaviour was noted by 95 per cent of owners and 59 per cent of the owners reported a noticeable change of behaviour within a month of surgery. Given the seasonal variation in testicular size, the best period for castrations in male European tortoises is shortly after brumation (April, May), when testes are proportionally smaller.

Phacoemulsification cataract surgery in the loggerhead turtle (Caretta caretta): surgical technique and outcomes in 10 cases.

Cataract removal using phacoemulsification was performed in 10 loggerhead turtles being rehabilitated after stranding. All turtles had significant systemic abnormalities and had cataracts either at the time of rescue or developed them during rehabilitation. Surgical difficulties encountered included an extremely soft globe that did not allow for a partial thickness corneal incision, incomplete topical anesthesia of the ocular surface with proparacaine, inability to dilate the pupil pharmacologically, a markedly shallow anterior chamber, a thick posterior capsular plaque in most cases, and difficulty creating a watertight closure with sutures. Minimal to no intraocular inflammation was seen in all cases following surgery, but all cases in which corneal sutures were used developed a marked inflammatory reaction surrounding the sutures and appeared uncomfortable until sutures were removed or sloughed. All turtles appeared to have improved vision following surgery and were eventually released. Two turtles were re-encountered a year or more after release. Both showed signs of normal growth and the ability to capture prey in the wild. This report documents successful outcomes associated with cataract surgery in loggerhead turtles, but also presents significant surgical challenges that should be taken into account before attempting this procedure in this species.

Single-incision, prefemoral bilateral oophorosalpingectomy without coelioscopy in an Indian star tortoise (Geochelone elegans) with follicular stasis.

A female Indian star tortoise (Geochelone elegans) was evaluated for anorexia and suspected coelomic masses. Clinical examination indicated follicular stasis. The tortoise was anesthetized and placed in dorsal left lateral recumbency for a right prefemoral approach. The ovaries and oviducts were exposed through the right prefemoral incision. The ovarian vasculature, the mesovarium, the oviduct, mesosalpinx and associated vasculature were ligated and transected. Bilateral oophorosalpingectomy was performed through the incision, without coelioscopy. Since then, the same procedure has been performed in five other tortoises, and all recovered well. These results suggest that bilateral oophorosalpingectomy, performed through a prefemoral incision, without coelioscopy, is a safe and practical approach for treating follicular stasis in tortoises.

The use of silver amalgam for treatment of shell ulcerations in turtles: 60 cases.

Shell ulcers were diagnosed based on clinical appearance in 60 turtles. Under general anaesthesia, surgical curettage was performed using a burr mounted on a micro-grinder. A calcium hydroxide paste was applied to the debrided lesions before the application of restorative material. Lesions were completely filled with rapidly hardening silver amalgam. Treated terrapins were hospitalised in dry containers and later were housed in tanks with controlled temperature 24 hours. Fillings were found spontaneously expulsed at various periods after placement. No signs of mercury poisoning or tissue reaction to the prosthesis were apparent.

Surgery and electrochemotherapy for the treatment of cutaneous squamous cell carcinoma in a yellow-bellied slider (Trachemys scripta scripta).

CASE DESCRIPTION: A 5-year-old female yellow-bellied slider (Trachemys scripta scripta) was referred for evaluation of a 2-month nonhealing ulcerated mass on the dorsal aspect of the neck. CLINICAL FINDINGS: The turtle was quiet, alert, and responsive, with a 2 × 1.5-cm ulcerated lesion on the neck. Signs of discomfort were observed during manipulation of the neck; no other abnormalities were detected during physical evaluation. TREATMENT AND OUTCOME: Following total body radiography and hematologic and serum biochemical analysis, the turtle was anesthetized and the mass was surgically removed. The excised tissue was submitted for histologic evaluation. A histopathologic diagnosis of squamous cell carcinoma (SCC) was made. Further surgical revision was not an option because of the extensive nature of the lesion; therefore, the tumor bed was treated with electrochemotherapy (ECT). Two sessions of ECT were performed with a 2-week interval between treatments. Electrochemotherapy involved intratumoral administration of bleomycin followed by trains of biphasic electric pulses. The treatment was well tolerated, and the turtle was disease free after 12 months. CLINICAL RELEVANCE: ECT resulted in good local control of SCC and should be considered as a possible postsurgical adjuvant treatment in reptiles with cutaneous tumors.

Single surgeon coelioscopic orchiectomy of desert tortoises (Gopherus agassizii) for population management.

Orchiectomy in chelonians is a challenging procedure, especially in large species with deep and elongated testes and extensive mesorchial attachments. Single surgeon coelioscopic orchiectomy was performed in seven adult desert tortoises (Gopherus agassizii), maintained at the Desert Tortoise Conservation Center (DTCC) in Las Vegas, for population management. Surgery was successfully conducted through a bilateral prefemoral approach via sequential vascular clip ligation and radiosurgery (monopolar/bipolar). Bipolar endoscopic forceps were considered indispensable due to the extensive mesorchial attachments and their close association with the kidney. A mechanical arm was effectively used to permit orchiectomy to be completed by a single surgeon. Six of seven animals recovered from anaesthesia. Necropsy demonstrated that the death of the other was unrelated to surgical complications. One animal experienced surgically significant haemorrhage, but still made a clinical recovery. The six tortoises were returned to the DTCC and, six months postoperatively, remain healthy. This small study suggests this minimally invasive technique is an effective method for bilateral orchiectomy in desert tortoises and might be preferable in large chelonians with elongated testes.

Surgical treatment of injuries caused by fishing gear in the intracoelomic digestive tract of sea turtles.

We report the surgical techniques used to remove accidentally ingested hooks and branchlines localized in different parts of the digestive tract of 129 loggerhead sea turtles Caretta caretta, together with the characteristics and localization of lesions, and final outcome related to their severity. Hooks were removed from the cervical esophagus via the ventral surface of the neck, while the supraplastron approach was performed for hooks wedged in the intracoelomic portion of the esophagus. An approach through the left axillary region was preferred for fishhooks in the stomach, while hooks and long branchlines in the intestine or pyloric area were removed by approaching the coelomic cavity through the right or left prefemoral fossa. The ingestion of fishhooks, and/or longlines, often induces severe injuries in the digestive tract that could lead to the death of the turtles, with the extent of damage engendered by lines often more severe than that caused by hooks, leading to strangulation, intussusception, and tears that require resection of long tracts of intestine. Spontaneous expulsion of hooks, even where possible, involves long waiting times, with the possible impairment of the turtle's clinical condition, and should be avoided when the line is evident or suspected. The development of diversified surgical techniques enabled us to approach the coelomic cavity with minimally invasive and easy-to-perform methods, and survival rates proved very satisfactory.

Laser therapy in a soft-shelled turtle (Pelodiscus sinensis) for the treatment of skin and shell ulceration. A case report.

Skin and shell diseases in aquatic turtles are often associated with several underlying causes. The presented case report describes aetiology including differential diagnoses, diagnostic procedures and therapy of a soft-shelled turtle (Pelodiscus sinensis) suffering from a septicaemic ulcerative dermatitis. Central aspect hereby is the positive curing effect of laser therapy on skin and shell lesions.

The use of negative pressure wound therapy following subcarapacial abscess excision in a tortoise.

A 42-year-old female Testudo graeca was referred with a five-month history of a non-healing carapacial wound. Previous treatment had involved long-term antibiotic therapy and conscious debridement of necrotic tissue. On presentation an extensive deep wound with purulent discharge was visible on the right side of the carapace. A computed tomographic scan was performed to assess the extent of the wound. This revealed a large carapacial deficit with two underlying soft tissue masses. Surgical debridement was performed under general anaesthesia resulting in an even larger defect. Because of the extensive nature of the deficit, negative pressure wound therapy was applied to aid wound healing. A negative pressure of approximately 120 mmHg was maintained and bandages were changed every third day. Wound healing progressed rapidly and a healthy granulation bed was formed within 16 days.

Absence of post-lesion reactive gliosis in elasmobranchs and turtles and its bearing on the evolution of astroglia.

In the mature mammalian and avian central nervous systems, neuronal destructions are followed by reactive gliosis, but data on other vertebrates are rather controversial. Mammals and birds belong to different amniote groups (Synapsida and Diapsida, respectively), but exhibit common general features in their glial architecture, mainly the predominance of astrocytes. Two vertebrate groups seem to be in special positions of glial evolution: turtles (Testudiniformes) and skates and rays (Batoidea). The purely ependymoglial system of turtles seems to be the simplest one among the extant amniotes. In skates and rays, true astrocytes are preponderant glial elements, in contrast to the other "anamniotes" (and even to reptiles). We investigated stab wounds by the immunohistochemical detection of GFAP in turtles (Trachemys-formerly Pseudemys-scripta elegans), a skate (Raja clavata) and rays (Dasyatis akajei and Torpedo marmorata). Sharks (Scyliorhinus canicula) as ependymoglia-predominated chondrichthyans, and-for positive controls-rats were also studied. In the elasmobranchs, other astroglial markers: glutamine synthetase and S100 protein were also applied. Neither turtles nor elasmobranchs presented considerable astroglial reactions. Critically surveying the former reports on different vertebrates, these results complete the picture that typical post-lesion reactive gliosis is confined to mammals and birds. Analysis of the astroglial systems from phylogenetic perspective suggests that the capability of forming glial demarcation and scar formation evolved independently in mammals and birds. Predominance of astrocytes is a necessary condition but not sufficient for reactive gliosis. The intense glial reactivity of mammals and birds may be attributed to their complex cerebralization.

Assessment of the effects of intramuscular administration of alfaxalone with and without medetomidine in Horsfield's tortoises (Agrionemys horsfieldii).

OBJECTIVE: To characterise four different intramuscular (IM) anaesthetic protocols, two with alfaxalone and two with alfaxalone in combination with medetomidine in terrestrial tortoises. STUDY DESIGN: Blinded, randomized, cross-over experimental study. ANIMALS: Nine healthy adult male Horsfield's tortoises (Agrionemys horsfieldii). METHODS: Each tortoise was randomly assigned to one of four different protocols: 1) 10 mg kg(-1) alfaxalone; 2) 10 mg kg(-1) alfaxalone + 0.10 mg kg(-1) medetomidine; 3) 20 mg kg(-1) alfaxalone; and 4) 20 mg kg(-1) alfaxalone + 0.05 mg kg(-1) medetomidine. During the experiment, the following variables were recorded: heart rate; respiratory rate; peripheral nociceptive responses; muscle strength; ability to intubate; palpebral, corneal and tap reflexes; and cloacal temperature. RESULTS: Protocols 1 and 2 resulted in moderate sedation with no analgesia, and moderate to deep sedation with minimal analgesia, respectively. Protocols 3 and 4 resulted in deep sedation or anaesthesia with variable analgesic effect; these two protocols had the longest total anaesthetic time and allowed intubation in 6/9 and 8/9 tortoises respectively. The total anaesthesia/sedation time produced by alfaxalone was significantly increased (p < 0.05) by the addition of medetomidine. There were no significant differences regarding time to plateau phase and duration of plateau phase. Baseline heart rate of 53 ± 6 beats minute(-1) decreased significantly (p < 0.05) with all protocols, and was lower (p < 0.05) in protocols 3 and 4. Heart rate increased after atipamezole administration, but the increase was transient. In two tortoises, extreme bradycardia with no cardiac activity for 10 minutes was observed with protocols 3 and 4. CONCLUSION AND CLINICAL RELEVANCE: Alfaxalone 10 and 20 mg kg(-1) IM can be used for sedation for non-painful procedures. Alfaxalone in combination with medetomidine can be used for deeper sedation or anaesthesia, but the observed respiratory and cardiovascular depression may limit its use.

Coelioscopic orchiectomy can be effectively and safely accomplished in chelonians.

Coelioscopic orchiectomy was performed in 27 male turtles (25 juvenile to adult red-eared sliders (Trachemys scripta elegans), one adult eastern painted turtle (Chrysems picta picta), and one juvenile male yellow-spotted Amazon River turtle (Podocnemis unifilis)). Orchiectomy was conducted under coelioscopic visualisation using ligation and transection of the mesorchium, or transection of the mesorchium with monopolar radiosurgical scissors. In 22 cases, bilateral orchiectomy was performed through a single incision; five turtles required bilateral incisions. All turtles recovered from anaesthesia. Nine turtles died within one year of surgery from conditions believed to be unrelated to surgery. One turtle was lost to follow-up. Seventeen turtles remain clinically healthy one to three years postoperatively. Coelioscopic orchiectomy provides a minimally invasive method for sterilisation of male chelonians and provides excellent visualisation during surgery. This technique is a useful model for the development of additional minimally invasive surgical techniques for chelonians.