Pancuronium Bromide for Chemical Immobilization of Adult Nile Crocodiles (Crocodylus niloticus): A Field Study.

(1) Background: Pancuronium bromide is a neuromuscular blocker used for immobilizing crocodiles that can be reversed with neostigmine. A recommended drug dose has only been established for saltwater crocodiles (Crocodylus porosus), mostly based on trials in juveniles and subadults. After trialing a dose recommendation in a small cohort of nine Nile crocodiles (Crocodylus niloticus), we developed and applied a new dose recommendation for large adult Nile crocodiles. (2) Methods: we trialed and adapted a pancuronium bromide (Pavulon 4 mg/2 mL) dose in Nile crocodiles originally established for saltwater crocodiles and applied the new dose for the immobilization of 32 Nile crocodiles destined for transport. Reversal was achieved with neostigmine (Stigmine 0.5 mg/mL). (3) Results: Nine crocodiles were included in the trial phase; the induction time was highly variable (average: 70 min; range: 20-143 min), and the recovery time was prolonged (average: 22 h; range: 50 min-5 days), especially in large animals after reversal with neostigmine. Based on these results, we established a dose-independent recommendation (3 mg pancuronium bromide and 2.5 mg neostigmine) for animals weighing ≥ 270 kg (TL ≥ ~3.8 m). When applied to 32 adult male crocodiles (BW range: 270-460 kg; TL range: 3.76-4.48 m), the shortest induction time was ~20 min and the longest ~45 min. (4) Conclusions: Pancuronium bromide and its antidote, neostigmine, are effective for the immobilization and reversal of adult male Nile crocodiles (TL ≥ 3.8 m or BW ≥ 270 kg) when given in a weight-independent fashion.

COMPLETE UNILATERAL MAXILLECTOMY IN A COHORT OF FIVE COLOMBIAN BOAS (BOA IMPERATOR).

Five unrelated adult Colombian boas (Boa imperator) presented with a 1- to 3-mon history of unilateral rostral swelling of the maxilla associated with a chronic rubbing against the enclosure's walls. Moderate to severe gingival inflammation and ulceration of the labial mucosa were present at the level of the swelling with tenderness to the touch. Radiography revealed osteolytic or proliferative lesions of the maxillary bone. Chronic maxillary osteomyelitis was diagnosed. Unilateral maxillectomy was performed on each animal under general anesthesia. Local anesthesia was also achieved by infiltrating lidocaine along the medial and lateral aspect of the maxillary gingiva and at the level of the maxillo-ectopterygoid joint. Using a lateral gingival approach, the maxillo-prefrontal, maxillary-palatine, and maxillo-ectopterygoid attachments were transected, and the maxillary bone removed. Histologic examination revealed pyogranulomatous stomatitis and osteomyelitis in all snakes, and presence of intralesional bacteria (n = 3 snakes). Gram-negative bacteria (Chryseobacterium indologenes and Proteus mirabilis) were cultured from the resected tissue of two snakes. One snake suffered from wound dehiscence 5 d postoperatively. All snakes were fed 15 d postoperatively and ingested dead mice without apparent difficulties. One snake was examined 2 mon and 1 yr after surgery, with no evidence of soft tissue or osseous infection and only minor facial scaring; all other snakes were lost to follow-up 15 d after surgery. Unilateral maxillectomy was performed in a cohort of five Colombian boas suffering from maxillary osteomyelitis. This surgical technique should be considered as an alternative to medical treatment in boid snakes.

Effects of a Single Opioid Dose on Gastrointestinal Motility in Rabbits (Oryctolagus cuniculus): Comparisons among Morphine, Butorphanol, and Tramadol.

The aim of this study was to evaluate and compare the effects of single doses of butorphanol, morphine, and tramadol on gastrointestinal motility in rabbits (Oryctolagus cuniculus) using non-invasive imaging methods, such as radiographic barium follow through and ultrasonographic contraction counts. Time-lapse radiographic and ultrasound examinations were performed before and after a single intramuscular dose of 5 mg kg-1 butorphanol, 10 mg kg-1 morphine, or 10 mg kg-1 tramadol. Pyloric and duodenal contraction counts by ultrasonography and radiographic repletion scores for the stomach and caecum were analysed using a mixed linear model. No significant effect was noted on ultrasound examinations of pyloric and duodenal contractions after administration of an opioid treatment. Morphine had a significant effect on the stomach and the caecum repletion scores, whereas butorphanol had a significant effect only on the caecum repletion score. Tramadol had no significant effect on the stomach or caecum repletion scores. The present findings suggest that a single dose of 5 mg kg-1 butorphanol or 10 mg kg-1 morphine temporarily slows gastrointestinal transit in healthy rabbits, preventing physiological progression of the alimentary bolus without the induction of ileus. In contrast, a single dose of 10 mg kg-1 tramadol has no such effects.

IRIDIUM 192 (192-Ir) HIGH DOSE RATE BRACHYTHERAPY IN A CENTRAL BEARDED DRAGON (POGONA VITTICEPS) WITH ROSTRAL SQUAMOUS CELL CARCINOMA.

A 0.5-kg, 9-yr-old, male central bearded dragon (Pogona vitticeps) presented with a proliferative mass (0.4 × 0.2 inches) on the left rostral aspect of the lower lip. Physical examination, blood work, and whole-body radiography did not reveal any other abnormalities. Histopathology confirmed squamous cell carcinoma. Considering the small size of the tumor, absence of deep tissue infiltration, and its radioresponsive characteristics, iridium 192 high dose rate brachytherapy was attempted. The dragon initially received three doses of 4 Gy/site at days 0, 7, and 17. Recurrence developed 3 mo later. Three more fractions of 6 Gy/site at days 0, 7, and 14 were delivered according to the same procedure. A second recurrence appeared after 2 mo. Surgical excision was then performed, followed by four fractions of 6 Gy/site on the surgical site at 2-wk intervals. Sixteen months posttreatment, no recurrence of the mass was observed.

TWO-DIMENSIONAL ECHOCARDIOGRAPHIC MEASUREMENTS IN THE BALL PYTHON (PYTHON REGIUS).

Reptiles can suffer from infectious and noninfectious cardiac pathologies, requiring the need for standardized diagnostic approaches and reference intervals. Despite the popularity of ball pythons (Python regius) as pets, echocardiographic measurements are unknown in this species. Twenty healthy adult ball pythons were evaluated to identify imaging planes, establish reference intervals for cardiac assessment by two-dimensional echocardiography, and study the effects of sex, body length, and body mass on heart rate, fractional shortening, and vascular, atrial, and ventricular dimensions. Echocardiography was performed under manual restraint. Most cardiac measurements were positively correlated with body length and mass, with the strongest correlation between ventricular end-systolic measurements and body length. The only significant difference found between sexes was for right and left atrial lengths. This study provides guidelines and reference intervals for two-dimensional echocardiographic measurements in adult healthy ball pythons.

HYPERTENSIVE HEART DISEASE AND ENCEPHALOPATHY IN A CENTRAL BEARDED DRAGON (POGONA VITTICEPS) WITH SEVERE ATHEROSCLEROSIS AND FIRST-DEGREE ATRIOVENTRICULAR BLOCK.

A 0.5 kg, 5-yr-old male bearded dragon (Pogona vitticeps) presented with a 2-mo history of lethargy, anorexia, and impaired locomotion. Upon physical examination, bradyarrhythmia (heart rate: 20 beats/min) and balance disorders were noted. Electrocardiography revealed a first-degree atrioventricular block (P-R interval: 360 ms). On echocardiography, all cardiac chambers were slightly above normal ranges. Complete blood count, blood biochemistry, and T4 were unremarkable except for mildly elevated aspartate aminotransferase. Adenovirus testing was negative by polymerase chain reaction. Following euthanasia, necropsy revealed marked thickening of the arterial trunks and histopathology confirmed multifocal atherosclerosis of efferent heart vessels, arteriosclerosis of cerebral arterioles, and multifocal spongiosis of brain tissue, more pronounced in the optic chiasma. Owing to its severity, atherosclerosis may have contributed to chronic arterial hypertension with damages to the heart, brain vessels, and brain tissue-optic chiasma.

In vitro and in vivo assessment of eprociclovir as antiviral treatment against testudinid herpesvirus 3 in Hermann's tortoise (Testudo hermanni).

Tortoises belonging to the Testudinidae family are infected by Testudinid herpesviruses. Testudinid herpesvirus 3 (TeHV-3) is considered the most pathogenic and affects several tortoise species, particularly those from the Testudo genus. As most species of this genus are endangered contribute to ecological concerns over this virus. Here, we aimed to explore the rational development of an antiviral treatment against TeHV-3 using Hermann's tortoise (Testudo hermanni) as a host model. Ten antiviral compounds were tested in cell culture for their toxicity and their activity against TeHV-3. Eight compounds exhibited different levels of activity against TeHV-3 with either no or only minor cytotoxic effects on cells. Next, eprociclovir (EPV, ciprovir) was selected for further investigations in vivo. Its pharmacokinetic properties were investigated after a single sub-cutaneous administration at 5 or 10 mg/kg. Plasma concentrations remained above half maximal effective concentration (EC50) for 2.2 and 4.4 h after administration at 5 and 10 mg/kg, respectively. Finally, EPV toxicity was investigated after administration at the dose of 10 mg/kg, BID for seven consecutive days. As early as one day after initiation of the treatment up to its end, EPV plasma concentration remained under the EC50. Apathy and anorexia developed after 7 days. Biochemical and anatomopathological examinations revealed nephrotoxic effects of EPV. Altogether, these data suggest that EPV is not a suitable molecule for the treatment of TeHV-3. Further studies are required to determine whether the other molecules identified here for their anti-TeHV-3 activity represent potential candidates for the development of efficacious treatments.

Influence of a single dose of buprenorphine on rabbit (Oryctolagus cuniculus) gastrointestinal motility.

OBJECTIVE: To establish a noninvasive imaging protocol for rabbit gastrointestinal transit evaluation. To assess the effect of a single injection of buprenorphine on the digestive transit of rabbits via this new technique. STUDY DESIGN: Prospective, parallel study. ANIMALS: Fifteen specific pathogen-free male New Zealand White rabbits weighing 2.68 ± 0.28 kg. METHODS: A 10 mL kg-1 barium meal was administered and the rabbits were subjected to serial radiographic and ultrasound examinations without treatment and 1 week later following a single intramuscular dose of 100 µg kg-1 of buprenorphine. Radiographic data from the stomach and caecum were collected and assigned a retention score ranging from 0 (no barium) to 3 (large amount of barium). The resulting scores and pyloric and duodenal contraction counts were analysed using a mixed linear model and are expressed as least square mean (lsm) ± standard error. Transit was estimated based on the apparition time of faeces in the pelvic area and analysed using a Wilcoxon test. A p < 0.05 was considered significant. RESULTS: Buprenorphine treatment induced a higher lsm number of pyloric (1.73 ± 0.19 versus 0.78 ± 0.19, p < 0.01) and lsm duodenal contractions (17.35 ± 1.04 versus 13.44 ± 1.04, p < 0.01). Buprenorphine administration decreased the lsm barium retention score in the stomach (2.44 ± 0.05 versus 2.64 ± 0.05, p < 0.01), but had no effect on the lsm barium retention score in the caecum. The time to apparition of faeces in the pelvic area was not influenced by buprenorphine administration (p = 0.66). CONCLUSIONS AND CLINICAL RELEVANCE: A single high dose of buprenorphine appears to have no adverse effect on gastrointestinal motility in healthy rabbits.

Toxicokinetics of selenium in the slider turtle, Trachemys scripta.

Selenium (Se) is an essential element that can be harmful for wildlife. However, its toxicity in poikilothermic amniotes, including turtles, remains poorly investigated. The present study aims at identifying selenium toxicokinetics and toxicity in juvenile slider turtles (age: 7 months), Trachemys scripta, dietary exposed to selenium, as selenomethionine SeMet, for eight weeks. Non-destructive tissues (i.e. carapace, scutes, skin and blood) were further tested for their suitability to predict selenium levels in target tissues (i.e. kidney, liver and muscle) for conservation perspective. 130 juvenile yellow-bellied slider turtles were assigned in three groups of 42 individuals each (i.e. control, SeMet1 and SeMet2). These groups were subjected to a feeding trial including an eight-week supplementation period SP 8 and a following 4-week elimination period EP 4 . During the SP8, turtles fed on diet containing 1.1 ± 0.04, 22.1 ± 1.0 and 45.0 ± 2.0 µg g(-1) of selenium (control, SeMet1 and SeMet2, respectively). During the EP4, turtles fed on non-supplemented diet. At different time during the trial, six individuals per group were sacrificed and tissues collected (i.e. carapace, scutes, skin, blood, liver, kidney, muscle) for analyses. During the SP8 (Fig. 1), both SeMet1 and SeMet2 turtles efficiently accumulated selenium from a SeMet dietary source. The more selenium was concentrated in the food, the more it was in the turtle body but the less it was removed from their tissues. Moreover, SeMet was found to be the more abundant selenium species in turtles' tissues. Body condition (i.e. growth in mass and size, feeding behaviour and activity) and survival of the SeMet1 and SeMet2 turtles seemed to be unaffected by the selenium exposure. There were clear evidences that reptilian species are differently affected by and sensitive to selenium exposure but the lack of any adverse effects was quite unexpected. Fig. 1 Design of the feeding trial. T, Time of tissues collection in weeks. The feeding trial included a supplementation period of 8 weeks (i.e. SP8) followed by an elimination period of 4 weeks (i.e. EP4). Six turtles from each turtle group (i.e. control, SeMet1 and SeMet2) were sacrifice at each collection time, from T1 to T12. At T0, four turtles were sacrificed.

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.

UNLABELLED: 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. IMPORTANCE: 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.

Detection of Usutu virus in a bullfinch (Pyrrhula pyrrhula) and a great spotted woodpecker (Dendrocopos major) in north-west Europe.

In October 2012, a 3-year-old bullfinch (Pyrrhula pyrrhula) held in captivity for its entire lifespan and a wild adult great spotted woodpecker (Dendrocopos major), both with neurological signs, were found 4 km from each other and 5 days apart in the Meuse Valley, Belgium. Non-suppurative encephalitis and mild degeneration and necrosis were identified in the brain and cerebellum, and Usutu virus antigen and RNA were detected by immunohistochemistry and real-time reverse transcriptase PCR, respectively. The two cases reported here represent the most western distribution of clinical disease in birds due to Usutu virus in Europe.