Comparison of Subcutaneous versus Intramuscular Dexmedetomidine-Midazolam-Ketamine-Morphine (DMKM) Mixture as Chemical Restraint for Endoscopic Sex Determination in Aldabra Giant Tortoises (Aldabrachelys gigantea).

Sex identification through coelioscopy is a minimally invasive surgical technique used to determine the sex of chelonians by directly visualizing their internal reproductive organs. An adequate anaesthesiologic plan is essential to guarantee patient immobilization and proper analgesia during the entire surgical procedure. In this study, we evaluated the effects of a combination of dexmedetomidine (0.05 mg/kg), midazolam (1 mg/kg), ketamine (8 mg/kg), and morphine (1 mg/kg) (DMKM) randomly delivered intramuscularly (IM) or subcutaneously (SC) in twenty-one Aldabra giant tortoise (Aldabrachelys gigantea) into the right antebrachium for celioscopic sex identification. Heart rate (HR), respiratory rate (RR), and body temperature (BT) were measured, along with the skeletal muscle tone of the thoracic and pelvic limbs, neck retraction reflex, palpebral reflex, and jaw tone every 15 min. The anaesthesiologic plan was considered to be adequate at the loss of the thoracic and pelvic limb retraction reflexes. After a 45 min interval, if the anaesthetic plan was deemed insufficient for the celioscopic procedure, a 5 mg/kg dose of propofol was administered intravenously into the subcarapacial venous plexus. At the end of the procedure, atipamezole (0.5 mg/kg) and flumazenil (0.05 mg/kg) were administered intramuscularly into the left antebrachium as reversal agents. Both HR and RR decreased from baseline to both 15 and 30 min. Due to the persistence of thoracic and pelvic limb retraction reflexes 45 min after DMKM administration, 6/11 (55%) cases in the SC group required the additional administration of propofol, in contrast to only 1/10 (10%) cases in the IM group (p = 0.05). The recovery times were comparable between the successfully induced animals in the IM and SC groups. In this study, the intramuscular administration of a DMKM combination quickly produced chemical restraint, suitable for celioscopic sex determination.

Sex identification in juvenile and adult Indonesian blue-tongued skinks (Tiliqua gigas) through cystoscopy and accuracy of contrast radiography.

OBJECTIVE: To describe a nonsurgical endoscopic technique for sex identification in Indonesian blue-tongued skinks (Tiliqua gigas) and to assess accuracy of contrast radiography of the hemipenile/hemiclitoral pouches. ANIMALS: 42 clinically healthy Indonesian blue-tongued skinks between 6 months and 3 years old and weighing between 22 and 550 g. METHODS: Cystoscopy was performed under general anesthesia. Gonads were visualized through the transparency of the urinary bladder, and their gross morphology was described. Contrast was applied in the tail pouches before obtaining full-body radiographs. Two radiologists, blinded to the sex of the skink, evaluated the radiographs. RESULTS: Cystoscopy was achieved in all 42 skinks. Visualization of the gonads through the urinary bladder was possible in 41 (98%; 95% CI, 87% to 99%) of the skinks, with 18 of them identified as males and 23 identified as females. Median procedure time was 60 seconds (range, 25 to 180 seconds) and was not associated with procedure order (-0.69; 95% CI, -1.83 to 0.45) or with the weight (0.02 g; 95% CI, -0.07 to 1.0) or the identified sex (11.7; 95% CI, -15.07 to 38.45) of the skink. Radiographs had a sensitivity of 69.6% (95% CI, 47.1% to 86.8%) and a specificity of 75.0% (47.6% to 92.7%) to identify female skinks. All the skinks recovered uneventfully. CLINICAL RELEVANCE: Cystoscopic sex identification is feasible in Indonesian blue-tongued skinks of various age and size. Considering the difficulty in identifying their sex otherwise, this technique could provide a significant improvement in the veterinary care of this species. In this population, contrast radiographs showed limited accuracy for sex identification.

First case of systemic fatal mycobacteriosis caused by Mycobacterium goodii in a pet Kenyan sand boa (Eryx colubrinus loveridgei).

BACKGROUND: Environmental nontuberculous mycobacteria species that are not members of the M. tuberculosis complex, are ordinary inhabitants of a wide variety of environmental reservoirs and their role in human and animal diseases has been fully recognized. Even if spontaneous mycobacterial infections have been reported in a wide variety of reptiles, this is the first report of systemic fatal mycobacteriosis sustained by Mycobacterium goodii in a pet reptile.  CASE PRESENTATION: An adult, wild caught (WC), male Kenyan sand boa (Eryx colubrinus loveridgei) age unknown, was presented for clinical examination due to decreased activity level, decreased appetite and diarrhea. Blood tests showed unreliable results. Coprologic exam showed a moderate to severe presence of flagellates. X rays and ultrasound showed moderate presence of air and faeces in the large intestine. The snake was hospitalized and oral metronidazole was chosen as antiprotozoal agent in association with subcutaneous warm fluids. The snake was discharged after 2 weeks therapy in good clinical condition. Faecal exam resulted negative. One month after, the snake was quickly hospitalized again because of a recrudescence of symptoms. Biochemistry showed severe increase of AST, ALT and biliary acids. Severe leucocytosis and moderate to severe anemia were highlighted. Ultrasound examination revealed a severe diffused alteration of the liver parenchyma and a fine needle aspiration was performed. The cytological diagnosis was mixed inflammation, with a numerous of unstained rod-shaped bacteria both inside macrophages and free in the sample. The snake's condition rapidly deteriorated and euthanasia was performed. The histology of the coelomic organs confirmed a systemic mycobacteriosis. Real-time PCR identified the mycobacteria as Mycobacterium goodii. CONCLUSIONS: Species from the genus Mycobacterium are among the most important micro-organism including the causative agents of tuberculosis. Even if the general incidence of disease in reptiles due to mycobacteria is comparatively low, they can serve as reservoirs of many ubiquitous mycobacteria species. Mycobacterium goodii is a rapidly growing non-tuberculous mycobacterium that has recently been associated with severe infections in animals and humans. Although in this case the pathogenesis was not completely clear, we highlight the zoonotic risk of mycobacteriosis in exotic animals especially in WC specimens.

Clinical, surgical, and pathological findings in client-owned rabbits with histologically confirmed appendicitis: 19 cases (2015-2019).

OBJECTIVE: To report clinical, surgical, and pathological findings in client-owned rabbits with histologically confirmed appendicitis. ANIMALS: 19 rabbits. PROCEDURES: Medical records for client-owned rabbits that had a histologic diagnosis of appendicitis were reviewed. RESULTS: Median age of the rabbits at presentation was 24.0 months (range, 4 to 84 months). Seventeen cases occurred during the summer and fall seasons. Decreased appetite (17/19 rabbits), abnormal rectal temperature (hyperthermia, 9/16 rabbits; hypothermia, 4/16 rabbits), hypocalcemia (8/11 rabbits), and hypoglycemia (7/15 rabbits) were common signs. Abdominal ultrasonography and CT findings were suggestive of appendicitis in 6 of 8 rabbits and in 1 of 2 rabbits, respectively. Of the 6 rabbits that received medical treatment, 3 died at 48 hours, 1 died at 24 hours after hospitalization, and 1 died at 10 days after presentation; 1 rabbit was alive at 1,030 days after presentation. Of the 8 rabbits that underwent appendectomy, 3 died before discharge from the hospital and 1 died 113 days after surgery; 4 rabbits were alive at 315, 334, 1,433, and 1,473 days after presentation. The remaining 5 rabbits either died or were euthanized before treatment could be instituted. In each of the 19 rabbits, the appendix had evidence of severe inflammation with mucosal ulceration, heterophilic inflammation, and necrotic debris. CLINICAL RELEVANCE: For rabbits with decreased appetite and an apparently painful abdomen, hyperthermia, hypocalcemia, or hypoglycemia, appendicitis should be considered as a differential diagnosis. Further comparisons of medical and surgical treatments are required to establish treatment recommendations for rabbits with appendicitis.

Lack of detection of West Nile virus in an islander population of chelonians during a West Nile virus outbreak.

In 2011, several outbreaks of West Nile disease occurred in Sardinia (Italy). The region hosts several chelonian species. Because of the increasing concern on the potential role that ectotherms may play in the ecology of West Nile virus (WNV), in October 2011 blood samples were collected from 41 endemic Sardinian chelonians and tested for the presence of active WNV infection or neutralizing antibodies by real time polymerase chain reaction (RT-PCR) and serumneutralisation, respectively. Neither WNV neutralising antibodies (0%; 95% CI: 0­8.4%) nor WNV RNA (0%; 95% CI: 0­6.8%) were found in the tested samples. According to the results of this screening survey, it is unlikely that chelonians are involved in the epidemiology of the 2011 WNV outbreaks in Sardinia.

PRELIMINARY EVALUATION OF SPIROTOME® DEVICE FOR LIVER BIOPSY IN GREEN IGUANAS (IGUANA IGUANA): A PILOT STUDY.

The aim of this study was to evaluate a large-core manual biopsy device (Spirotome(®), Medinvents, 3500 Hasselt, Belgium) for liver sampling and histologic diagnosis in green iguanas (Iguana iguana). The study included eight green iguanas, and two ultrasound-guided biopsies were collected for each lizard, for 16 biopsies in total. The procedure was carried out under general anesthesia induced by intravenous injection of propofol (10 mg/kg) maintained with a mixture of 2.0% isoflurane and 0.8-1.2 L/min oxygen after tracheal intubation. Fourteen (87.5%) of the 16 biopsies were considered diagnostic. Liver biopsy quality was assessed according to sample size and tissue preservation. In particular, mean length (16.2 ± 4.5 mm), width (2.2 ± 0.5 mm), area (34.8 ± 6.9 mm(2)), and number of portal areas (9.4 ± 3.9) of each biopsy were recorded for all green iguanas. The total available surface of the sections obtained from the biopsies and their grade of preservation enabled a satisfactory evaluation of the parenchymal architecture. One of the green iguanas in the study died the day after the procedure due to severe hemocoeloma. Risk assessment evaluation suggested that small green iguanas may not be suitable for this biopsy procedure.

Use of Cystoscopy to Visualize Morphological Alteration of the Liver in a Posthatchling Turtle ( Cuora trifasciata ).

A 55 g, 7 mo old Chinese three-striped box turtle ( Cuora trifasciata ) was presented with a 2 wk history of lethargy and anorexia. The owner attempted various antibiotic treatments without clinical improvement. A cystoscopic evaluation of the coelom was performed. The liver appeared dark red-to-brown presenting multifocal irregularly shaped white-to-tan and roughly round areas of discoloration on the capsular surface. An adult keeled box turtle (Pyxidea mouhotii), which died in the same facility at the same time, had similar hepatic lesions detected upon necropsy. From hepatic lesions, an antibiotic-resistant Escherichia coli was isolated. Antibiotic treatment based on sensitivity testing led to a rapid resolution of the clinical signs in the Cuora trifasciata , with gain of appetite 4 days after the first antibiotic administration. A recheck cystoscopy demonstrated macroscopic resolution of hepatic lesions. This report demonstrates, to the best of the authors' knowledge, for the first time the clinical usefulness of cystoscopy for the diagnostic evaluation of the coelom of small chelonians. This minimally invasive technique is a valuable clinical tool in the medical approach to posthatchling chelonians.

MITE INFECTION IN A MASKED PALM CIVET (PAGUMA LARVATA) TREATED BY SELAMECTIN (STRONGHOLD®, PFIZER LTD.).

The masked palm civet (Paguma larvata) is a small Asian mammal (order Carnivora, family Viverridae) uncommon in Italy. Limited information is available about management and sanitary maintenance in captivity. A 4-mo-old masked palm civet presented with pruritus, itch, scratching, and disorexia. On physical examination, alopecia and crusts were detected on the ventral and lateral trunk, tail, legs, and lips. Skin scrapings and cytology revealed Notoedres spp. and bacterial infection. On histopathology, parasitic dermatitis was observed with the presence of a Sarcoptidae mite and Demodex spp. Selamectin spot-on (15 mg/kg every 2 wk, three applications) and marbofloxacin per os (2.5 mg/kg once daily for 2 wk) were administered, and the animal recovered in 1 mo. With the good response to this therapy, a notoedric mange was thought to be the main problem. This is the first report about the use of selamectin to treat a mite infection in masked palm civet.

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.

Pharmacokinetics of cefovecin sodium after subcutaneous administration to Hermann's tortoises (Testudo hermanni).

OBJECTIVE: To determine the pharmacokinetics of cefovecin sodium after SC administration to Hermann's tortoises (Testudo hermanni). ANIMALS: 23 healthy adult Hermann's tortoises (15 males and 8 females). PROCEDURES: Cefovecin (8.0 mg/kg) was injected once in the subcutis of the neck region of Hermann's tortoises, and blood samples were obtained at predetermined time points. Plasma cefovecin concentrations were measured via ultraperformance liquid chromatography coupled to tandem mass spectrometry, and pharmacokinetic parameters were calculated with a noncompartmental model. Plasma protein concentration was quantified, and the percentage of cefovecin bound to protein was estimated with a centrifugation technique. RESULTS: Cefovecin was absorbed rapidly, reaching maximum plasma concentrations between 35 minutes and 2 hours after administration, with the exception of 1 group, in which it was reached after 4 hours. The mean ± SD time to maximum concentration was 1.22 ± 1.14 hours; area under the concentration-time curve was 220.35 ± 36.18 h•µg/mL The mean protein-bound fraction of cefovecin ranged from 41.3% to 47.5%. No adverse effects were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of a single dose of cefovecin SC appeared to be well-tolerated in this population of tortoises. Results of pharmacokinetic analysis indicated that the 2-week dosing interval suggested for dogs and cats cannot be considered effective in tortoises; however, further research is needed to determine therapeutic concentrations of the drug and appropriate dose ranges.

Disseminated mycobacteriosis in a stranded loggerhead sea turtle (Caretta caretta).

A loggerhead sea turtle (Caretta caretta) was found stranded alive along the Adriatic coast close to Ancona, Italy, displaying obtundation, tachypnea, and increased respiratory effort. It died a few hours after admission, and a postmortem examination was immediately performed. Miliary yellowish nodules were evident in the liver, and a lower number in the heart, stomach, and gut wall. Hundreds of whitish nodules were scattered in the lungs, with the majority of the pulmonary parenchyma being replaced by the lesions. Histologically, all nodular lesions consisted of a small central area of necrosis with acid-fast bacilli surrounded by epithelioid cells, macrophages, and lymphocytes. Giant cells were found in the spleen and the liver. Kidneys, lungs, liver, spleen, brain, and skin lesions were inoculated aseptically onto general isolation media and selective isolation media for mycobacteria. The isolate showed a restriction pattern identical to Mycobacterium chelonae by polymerase chain reaction-restriction fragment length polymorphism. To the best of the authors' knowledge, this is the first description of a disseminated infection caused by a potentially pathogenic mycobacteria in a stranded, free-ranging loggerhead sea turtle. Veterinary staff and biologists who handle sea turtles with suspected mycobacterial disease should protect themselves appropriately.

Evaluation of liver parenchyma and perfusion using dynamic contrast-enhanced computed tomography and contrast-enhanced ultrasonography in captive green iguanas (Iguana iguana) under general anesthesia.

BACKGROUND: Contrast-enhanced diagnostic imaging techniques are considered useful in veterinary and human medicine to evaluate liver perfusion and focal hepatic lesions. Although hepatic diseases are a common occurrence in reptile medicine, there is no reference to the use of contrast-enhanced ultrasound (CEUS) and contrast-enhanced computed tomography (CECT) to evaluate the liver in lizards. Therefore, the aim of this study was to evaluate the pattern of change in echogenicity and attenuation of the liver in green iguanas (Iguana iguana) after administration of specific contrast media. RESULTS: An increase in liver echogenicity and density was evident during CEUS and CECT, respectively. In CEUS, the mean ± SD (median; range) peak enhancement was 19.9% ± 7.5 (18.3; 11.7-34.6). Time to peak enhancement was 134.0 ± 125.1 (68.4; 59.6-364.5) seconds. During CECT, first visualization of the contrast medium was at 3.6 ± 0.5 (4; 3-4) seconds in the aorta, 10.7 ± 2.2 (10.5; 7-14) seconds in the hepatic arteries, and 15 ± 4.5 (14.5; 10-24) seconds in the liver parenchyma. Time to peak was 14.1 ± 3.4 (13; 11-21) and 31 ± 9.6 (29; 23-45) seconds in the aorta and the liver parenchyma, respectively. CONCLUSION: CEUS and dynamic CECT are practical means to determine liver hemodynamics in green iguanas. Distribution of contrast medium in iguana differed from mammals. Specific reference ranges of hepatic perfusion for diagnostic evaluation of the liver in iguanas are necessary since the use of mammalian references may lead the clinician to formulate incorrect diagnostic suspicions.

Computed tomography-guided bone biopsies for evaluation of proliferative vertebral lesions in two boa constrictors (Boa constrictor imperator).

Two boa constrictors (Boa constrictor imperator) presented with paresis of the trunk originating cranial to the cloaca. Radiographs were consistent with proliferative bone lesions involving several vertebrae. Computed tomography (CT) demonstrated the presence of lytic/expansile lesions. Computed tomography-guided biopsies of the lesions were performed without complications. Histology was consistent with bacterial osteomyelitis and osteoarthritis. Gram-negative bacteria (Salmonella sp. and Pseudomonas sp.) were isolated from cultures of the biopsies. Medical treatment with specific antibiotics was attempted for several weeks in both cases without clinical or radiographic improvements. The animals were euthanized, and necropsy confirmed the findings observed upon CT. To the authors' knowledge, this is the first report of the use of CT-guided biopsies to evaluate proliferative vertebral lesions in snakes. In the present report, CT-guided biopsies were easily performed, and both histologic and microbiologic results were consistent with the final diagnosis.

Clinical hematology in reptilian species.

The presence of nucleated erythrocytes and thrombocytes in reptile blood limits the use of electronic cell-counting devices for leukocytes so that manual counting techniques and evaluation of blood smears are necessary, resulting in time-consuming procedures requiring good training and experience. The evaluation of most hematologic values is complicated by the many physiologic adaptations occurring in reptiles, making references values very difficult to interpret. This article brings together most of the bibliography about hemochrome evaluation in reptiles, with schematic instruction about sample collection, handling, and evaluation. Information about evaluation of both qualitative and quantitative aspects of reptile hematology are also given.

Fire ignition during laser surgery in pet rodents.

BACKGROUND: Laser surgery is an attractive alternative to other means of section device in terms of tissue inflammation and interaction, which has been extensively used in human and veterinary medicine. Although accidental ignition during laser surgeries is sporadically reported in human medical literature, to the authors' knowledge this is the first report regarding laser-dependent fire ignition during surgery in veterinary medicine. CASE PRESENTATION: Two rodents, a 13-month old, 27-gram, male pet mouse (Mus musculus) and a 1-year old, female Russian hamster (Phodopus sungorus), underwent surgical removal of masses with diode laser. During the surgical procedures fires ignited from the face masks. The mouse presented severe burns on the head and both forelimbs, it was hospitalized and approximately 2 months after surgery burns were resolved. The hamster presented severe burns on the face and the proximal regions of the body. At 72 hours from the accident the hamster was euthanized. CONCLUSION: The present report suggests that fire ignition is a potential life-threatening complication of laser surgery in non-intubated rodents maintained under volatile anesthesia. High oxygen concentrations, the presence of combustible, and the narrowness of the surgical field with the face mask during laser surgery on rodents are risk factors for fire ignition.

Cutaneous progressive angiomatosis on the muzzle of a dog, treated by laser photocoagulation therapy.

A 10-year old, female, cocker spaniel was presented with a bleeding lesion on the muzzle and nasal planum observed initially as a small raised nodule at 1 year of age. This became clinically more pronounced at 5 years of age, progressively enlarged thereafter and was diagnosed initially as a vascular tumour. On clinical examination, multiple confluent nodules of variable diameter (0.5-1.5 cm) deformed the outline of the nose. Histological examination revealed a progressive angiomatosis with multifocal infiltrative growth of increased dermal vascularization with different sized and type of anastomosing vessels lined by a monolayer of reactive endothelium, often arranged as papillary projections into the lumina. Partial nosectomy, the first therapeutic approach, was unsuccessful and the lesion relapsed within 6 months. However, subsequent laser photocoagulation therapy provided a good cosmetic outcome, no relapse 1 year later, and appeared to provide an effective alternative to ablation surgery.

Osteitis deformans (Paget's disease) in a Burmese python (Python molurus bivittatus)--a case report.

Osteitis deformans (Paget's disease of bone) is a chronic focal disorder of bone remodelling characterized by an initial increase in osteoclast-mediated bone resorption, with subsequent compensatory increase in new bone formation, resulting in a disorganized mosaic of woven and lamellar bone. In the Burmese python (Python molurus bivittatus) of this report, multifocal gross swellings involving the proximal third of the vertebral spine were observed and associated with anorexia, a relative inability to move or to fully extend the body, and to strike at prey. Serum biochemistry revealed elevated alkaline-phosphatase activity. Radiographic changes (irregular bone proliferation along the vertebral margins), computed tomography scan results (abnormal mineral density), and histopathological features (generalized thickening of the bony trabeculae at the expense of the intertrabecular spaces and irregular patches of lamellar bone with a characteristic "mosaic" pattern) indicated osteitis deformans.