Three-dimensional genome architecture persists in a 52,000-year-old woolly mammoth skin sample.

Analyses of ancient DNA typically involve sequencing the surviving short oligonucleotides and aligning to genome assemblies from related, modern species. Here, we report that skin from a female woolly mammoth (†Mammuthus primigenius) that died 52,000 years ago retained its ancient genome architecture. We use PaleoHi-C to map chromatin contacts and assemble its genome, yielding 28 chromosome-length scaffolds. Chromosome territories, compartments, loops, Barr bodies, and inactive X chromosome (Xi) superdomains persist. The active and inactive genome compartments in mammoth skin more closely resemble Asian elephant skin than other elephant tissues. Our analyses uncover new biology. Differences in compartmentalization reveal genes whose transcription was potentially altered in mammoths vs. elephants. Mammoth Xi has a tetradic architecture, not bipartite like human and mouse. We hypothesize that, shortly after this mammoth's death, the sample spontaneously freeze-dried in the Siberian cold, leading to a glass transition that preserved subfossils of ancient chromosomes at nanometer scale.

Chromosome-length genome assemblies and cytogenomic analyses of pangolins reveal remarkable chromosome counts and plasticity.

We report the first chromosome-length genome assemblies for three species in the mammalian order Pholidota: the white-bellied, Chinese, and Sunda pangolins. Surprisingly, we observe extraordinary karyotypic plasticity within this order and, in female white-bellied pangolins, the largest number of chromosomes reported in a Laurasiatherian mammal: 2n = 114. We perform the first karyotype analysis of an African pangolin and report a Y-autosome fusion in white-bellied pangolins, resulting in 2n = 113 for males. We employ a novel strategy to confirm the fusion and identify the autosome involved by finding the pseudoautosomal region (PAR) in the female genome assembly and analyzing the 3D contact frequency between PAR sequences and the rest of the genome in male and female white-bellied pangolins. Analyses of genetic variability show that white-bellied pangolins have intermediate levels of genome-wide heterozygosity relative to Chinese and Sunda pangolins, consistent with two moderate declines of historical effective population size. Our results reveal a remarkable feature of pangolin genome biology and highlight the need for further studies of these unique and endangered mammals.

Use of oral chemotherapeutic agent, chlorambucil, as palliative treatment of thryoid adenocarcinoma in a golden-headed lion tamarin (Leontopithecus chrysomelas).

This case report describes the use of chlorambucil in a 7.5-year-old golden-headed lion tamarin (Leontopithecus chrysomelas) as palliative therapy for thyroid adenocarcinoma. Treatment was initiated at 0.1 mg/kg orally once daily. No physical abnormalities or substantial changes in complete blood cell counts and thyroid hormone levels from serial samples were detected.

Surgical and medical management for fractures of the second through fifth metacarpals in a red ruffed lemur (Varecia rubra).

A 21-yr-old female red ruffed lemur (Varecia rubra) was presented with swelling and disuse of the right manus. Severely displaced fractures of metacarpals II-V were diagnosed radiographically. The fractures were surgically stabilized with intramedullary Kirschner wires attached externally with an acrylic external fixator and a bone plate on the dorsal aspect of metacarpal III. The fractures of metacarpals II-V were predominantly healed on radiographs obtained 12 wk after surgery. However, diffuse disuse osteopenia and phalangeal contracture were present, with possible osteomyelitis. An exercise regimen of the affected hand was initiated due to the incomplete extension of the phalanges. After 4 wk of therapy, the extension of the phalanges had improved and the fractures appeared radiographically to be nearly completely healed. Although metacarpal fractures are common in nonhuman primates, they are reported infrequently in the literature.

Cutaneous atypical mycobacteriosis in a clouded leopard (Neofelis nebulosa).

A 16-yr-old male clouded leopard (Neofelis nebulosa) was presented for lethargy and anorexia. A cutaneous abdominal mass extending from the pubis to just caudal to the xiphoid process was present. A biopsy revealed histologic lesions consistent with an atypical mycobacterial infection consisting of diffuse, severe, pyogranulomatous dermatitis and panniculitis, with clear vacuoles and 3-5 microm, intravacuolar, faintly eosinophilic, filamentous bacilli that stained positively with FiteFaraco modified acid-fast stain. The clouded leopard had biochemical findings suggestive of chronic renal failure and euthanasia was elected. Histological evaluation of tissues collected at postmortem examination revealed multicentric B-cell lymphoma involving the oral cavity, liver, spleen, and multiple lymph nodes, bilateral testicular seminomas, thyroid follicular cell adenoma, thyroid C cell adenoma, and biliary cystadenomas. Bacterial culture and molecular sequencing identified the causative agent of the cutaneous abdominal mass as belonging to the Mycobacterium fortuitum group.

Architecture of the bronchial tree in Cuvier's dwarf caiman (Paleosuchus palpebrosus).

We imaged the lungs of five Cuvier's dwarf caiman (Paleosuchus palpebrosus) via computed tomography (CT) and micro-computed tomography (µCT) and compared these data to the lungs of the American alligator (Alligator mississippiensis). These data demonstrate anatomical commonalities between the lungs of P. palpebrosus and A. mississippiensis, and a few notable differences. The structural similarities are (a) a proximally narrow, distally widened, hook-shaped primary bronchus; (b) a cervical ventral bronchus that branches of the primary bronchus and immediately makes a hairpin turn toward the apex of the lung; (c) a sequential series of dorsobronchi arising from the primary bronchus caudal to the cervical ventral bronchus; (d) intraspecifically highly variable medial sequence of secondary airways; (e) sac-like laterobronchi; and (f) grossly dead-ended caudal group bronchi in the caudal and ventral aspects of the lung. The primary differences between the two taxa are in the overall number of large bronchi (fewer in P. palpebrosus), and the number of branches that contribute to the cardiac regions. Imaging data of both a live and deceased specimen under varying states (postprandial, fasting, total lung capacity, open to atmosphere) indicate that the caudal margin and position of the lungs shift craniocaudally relative to the vertebral column. These imaging data suggest that the smooth thoracic ceiling may be correlated to visceral movement during ventilation, but this hypothesis warrants validation. These results provide the scaffolding for future comparisons between crocodilians, for generating preliminary reconstructions of the ancestral crocodilian bronchial tree, and establishing new hypotheses of bronchial homology across Archosauria.

Esophageal diverticula in Parma wallabies (Macropus parma).

Four adult, wild caught Parma wallabies (Macropus parma) presented with intermittent, postprandial, midcervical swellings. Esophageal diverticula were discovered in the four animals. One of two wallabies was managed successfully with surgery. A third animal died of other causes. The fourth animal died with possible complications from the diverticulum. This is the first published report of esophageal diverticula in macropods.

Magnetic resonance imaging findings in a red kangaroo (Macropus rufus) with otitis.

Magnetic resonance imaging (MRI) was performed on an adult, male Red kangaroo (Macropus rufus) with a history of nonspecific neurologic signs and acute discharge from the left ear. MRI revealed findings consistent with otitis and possible osteomyelitis of the temporal and mastoid bones. To the authors' knowledge, this is the first report of otitis and MRI findings in a kangaroo.

Ibex-associated malignant catarrhal fever in a bongo antelope (Tragelaphus euryceros).

A 4-yr-old male bongo antelope (Tragelaphus euryceros) died after an acute clinical course involving a febrile illness, anorexia, lethargy, minor oculonasal discharge, and diarrhea. Histologic lesions were compatible with malignant catarrhal fever (MCF). Polymerase chain reaction (PCR) revealed an amplified region of a herpesviral DNA polymerase gene sequence nearly identical to that of a MCF virus previously identified in Nubian ibex (Capra nubiana). The bongo had been housed across from an exhibit containing Nubian ibex that tested positive for MCF viral antibodies by competitive inhibition enzyme-linked immunosorbent assay. Further testing of the zoo's ibex via PCR also revealed viral DNA sequences nearly identical to those found in the bongo's tissues.

Oral eosinophilic granulomas in tigers (Panthera tigris)--a collection of 16 cases.

Oral eosinophilic granulomas were diagnosed in 16 tigers (Panthera tigris). All lesions were located on the hard or soft palate and typically consisted of flat or slightly raised circular ulcers. Histologic features of these lesions were essentially identical to those seen in oral eosinophilic granulomas of domestic cats and dogs. No clinical signs were noted in eight cases, though various degrees of inappetence, excessive salivation, and dysphagia were noted in the other eight tigers. Six cases were not treated. Treatment for the remaining 10 cases centered on corticosteroids and additional treatments included surgical removal, cryotherapy, antibiotics, and chlorpheniramine. Treatment with corticosteroids did appear to be effective in some cases, though lesions would worsen after cessation of therapy and no cases were cured. In addition, three cases developed complications possibly related to this corticosteroid therapy. The etiology of these lesions remains unknown, though an underlying allergic condition is likely.

Epitheliotropic lymphoma in a captive bontebok (Damaliscus pygargus).

A 16-yr-old, female, captive-bred bontebok (Damaliscus pygargus) estimated to be 75 kg, was presented with a 1-mo history of chronic diarrhea and weight loss. A cause for the deteriorating condition was not found antemortem, and the bontebok was humanely euthanatized when it failed to respond to empirical therapy. On necropsy, numerous nodules were present on the pleural surfaces and within the lung parenchyma, the pancreas appeared inflamed and fibrotic. Histologic examination revealed lymphoma in the mesenteric lymph nodes, alimentary tract, reproductive tract, and liver. In the small and large intestine, rumen, esophagus, bile duct, and vagina, the neoplastic cells had a tropism for epithelium. The neoplastic cells within the epithelium and subepithelium of the alimentary tract stained positively for CD3, consistent with T cell differentiation.

Preliminary single-dose pharmacokinetics of marbofloxacin in ball pythons (Python regius).

Pharmacokinetics of marbofloxacin in two male and four female adult ball pythons (Python regius) was determined after i.v. and p.o. administration of a single dose. Using a crossover design, each snake was given a single 10 mg/kg dose of marbofloxacin i.v. and p.o. Blood samples were collected prior to and 0.5, 1, 1.5, 3, 6, 12, and 24 hr after marbofloxacin administration. Marbofloxacin was quantitated by use of liquid chromatography-mass spectrometry. Following p.o. administration, marbofloxacin had a peak plasma concentration (Cmax) of 9.40 microg/ml and a time to Cmax (Tmax) of 9.0 hr. Based on the plasma pharmacokinetics generated in this study and pending any further studies to evaluate potential toxicity and multi-dose pharmacokinetics, we suggest a dosage for marbofloxacin in ball pythons of 10 mg/kg p.o. at least every 48 hr, depending on the sensitivity of the pathogen and as a basis for further research.

Serologic responses after vaccination of fennec foxes (Vulpes zerda) and meerkats (Suricata suricatta) with a live, canarypox-vectored canine distemper virus vaccine.

Fennec foxes (Vulpes zerda) and meerkats (Suricata suricatta) are considered to be susceptible to canine distemper virus (CDV) infection. Although no definitive clinical cases of natural CDV infections have been reported, mortalities due to CDV have been suspected and are reported in other closely related species. A commercially available monovalent, live, canarypox-vectored CDV vaccine induced neutralizing antibody titers that were maintained for at least a year in both fennec foxes and meerkats.

Pharmacokinetics and tissue concentrations of azithromycin in ball pythons (Python regius).

OBJECTIVE: To determine pharmacokinetics and tissue concentrations of azithromycin in ball pythons (Python regius) after IV or oral administration of a single dose. ANIMALS: 2 male and 5 female ball pythons. PROCEDURES: Using a crossover design, each snake was given a single dose of azithromycin (10 mg/kg) IV. After a 4-week washout period, each snake was given a single dose of azithromycin (10 mg/kg) orally. Blood samples were collected prior to dose administration and 1, 3, 6, 12, 24, 48, 72, and 96 hours after azithromycin administration. Azithromycin was quantitated by use of liquid chromatography-mass spectrometry. RESULTS: After IV administration, azithromycin had an apparent volume of distribution of 5.69 L/kg and a plasma clearance of 0.19 L/h/kg. Harmonic means for the terminal half-life were 17 hours following IV administration and 51 hours following oral administration. Mean residence times were 37 and 94 hours following IV and oral administration, respectively. Following oral administration, azithromycin had a peak plasma concentration (Cmax) of 1.04 microg/mL, a time to Cmax of 8.4 hours, and a prolonged mean absorption time of 57 hours. Mean oral bioavailability was 77%. Tissue concentrations ranged from 4 to 140 times the corresponding plasma concentration at 24 and 72 hours after azithromycin administration. CONCLUSIONS AND CLINICAL RELEVANCE: Azithromycin is well absorbed and tolerated by ball pythons. On the basis of plasma pharmacokinetics and tissue concentration data, we suggest an azithromycin dosage in ball pythons of 10 mg/kg, orally, every 2 to 7 days, depending upon the site of infection and susceptibil ity of the infective organism.

Hematology and plasma biochemistry of captive puna ibis (Plegadis ridgewayi).

Normal hematology and plasma biochemistry values are reported for a single captive population of 14 puna ibises (Plegadis ridgewayi). The natural biology and conservation status of this species is virtually unknown and it is considered a vulnerable species in parts of South America. The values presented here may be useful to clinicians or wildlife managers working with puna ibises.

Assessment of multiple cardiocentesis in ball pythons (Python regius).

This study evaluated the gross and microscopic effects of serial blood collection from six ball pythons (Python regius) by using cardiocentesis. We collected 39 blood samples from each snake over a 120-day period. Cardiocentesis was performed on manually restrained snakes, with each sample requiring approximately 15 sec to collect. No clinically apparent complications were noted in any of the snakes after the cardiocentesis procedures, and all snakes survived until they were euthanized 73 days after the last blood sample. Minimal gross lesions were noted at necropsy; faint brown pigmentation of the pericardium was present in five of six snakes, and three snakes had approximately 0.5 ml dark pigmented fluid in the pericardial space. One snake had a small, organized hematoma in the pericardial space. Microscopic findings were limited to moderate and regularly arranged collagen fibrosis and focal thickening of the epicardium. The pericardial sac in all snakes had a mild infiltrate of hemosiderin-laden macrophages and small numbers of heterophils. The results suggest that serial cardiocentesis is well tolerated in ball pythons.

Ocular biology and diseases of Old World chameleons.

For centuries, Old World chameleons (Chamaeleonidae family) have been collected and studied for their unusual biology and features, which are unique among lizards and other vertebrates. They have advanced mechanisms for capturing prey with their tongue, but have a primitive mechanism for hearing. Chameleons have the most studied ocular system because of their highly adapted yet primitive biology. This system has specific features that are susceptible to new diseases, which may require novel therapies.

Dilated cardiomyopathy and amebic gastritis in a giant anteater (Myrmecophaga tridactyla).

An approximately 11-mo-old female giant anteater (Myrmecophaga tridactyla) exhibited anorexia, lethargy, hypothermia, depression, and minimal response to external stimuli. Radiography and ultrasonography revealed an enlarged heart, with free gas and fluid in the abdomen. Abdominocentesis produced a clear brown fluid with an acute to subacute septic suppurative exudate. Cardiac ultrasonography revealed a dilated, thin-walled left ventricle with a comparatively low fractional shortening. Despite intensive supportive care, the anteater died. Postmortem findings included gastric ulceration with perforation near the pylorus. Entameba spp. and Acanthamoeba spp. were both identified in large numbers at the site of the gastric ulceration and perforation.

Spinal compression due to atlantal vertebral malformation in two African lions (Panthera leo).

Two young (14-mo-old and 6-mo-old), unrelated, male African lions (Panthera leo) were presented to the Veterinary Teaching Hospitals of Oklahoma State University and Kansas State University with progressive ambulatory difficulty. In both cases, limited neurologic evaluation demonstrated pelvic limb paresis and ataxia with conscious proprioceptive deficits. Spinal imaging showed nearly identical lesions in both cases. Radiography and myelography demonstrated cervical stenosis secondary to atlantal (C1) malformation producing a dorsoventral deformity of the laminar arch with atlantoaxial spinal cord compression between the ventrally displaced laminar arch of the atlas and the underlying odontoid process of C2. Computed tomography of the atlanto-axial junction confirmed cervical stenosis and cord compression, showing flattening of the spinal cord between the laminar arch of C1 and the dens of C2. Decompressive surgery consisting of dorsal laminectomy of C1 was performed. Each lion demonstrated progressive improvement of neurologic status to recovery of normal ambulation after surgical intervention. Neurologic disease in large captive felids is rare; atlanto-axial spondylomyelopathy has not been reported previously.