The prevalence of antibiotic resistance in 376 Escherichia coli (E. coli) isolates from fecal samples of Hooded and White-naped cranes was investigated on the Izumi plain in Kagoshima prefecture, Japan, during winter 2016 and 2017. Resistance to oxytetracycline, ampicillin, and nalidixic acid were observed in 10.9%, 3.1–4.4%, and 2.1–7.7% of isolates, respectively. Since the previous surveillance in 2007, isolation rates of antibiotic-resistant E. coli recovered from wild cranes have remained at significantly low levels compared with those in Japanese livestock. Our results indicate that surveillance of antibiotic-resistant E. coli from wild cranes wintering in the Izumi plain could be a useful strategy to indicate natural environmental pollution by antibiotic-resistant bacteria in the environment.
Intestinal smooth muscle hypertrophy and hyperplasia has been described in human and several mammal species. In birds, only one case of intestinal smooth muscle hyperplasia has been reported. This paper describes the anatomopathological and histological findings of three cases of intestinal smooth muscle hypertrophy/hyperplasia in two different avian species belonging to the family Gallinidae and Columbidae. Grossly, it involved all tracts of the small intestine. Histologically, hyperplasia involved the mucosal villi, muscularis mucosa and inner and outer layers of the tunica muscularis. Hypertrophy was apparently detected only in the inner circular muscle layer. Lack of submucosal plexuses was also observed in all three animals. The results confirm the remarkable histological difference between mammals and avian species and show as these pathological changes can occur in different species of birds.
Actinobacillus species are known to be pathogenic to horses. To clarify etiological agents of actinobacillosis in Japanese adult horses, 27 isolates from Japanese Thoroughbred racehorses putatively identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry as Actinobacillus were further identified by PCR of the A. equuli toxin gene, by CAMP test, and by 16S rRNA sequencing analysis. Actinobacillus equuli subsp. haemolyticus was isolated most frequently (16/27) and was related to respiratory infections. Actinobacillus equuli subsp. equuli (4/27) was isolated from chronic cases or concomitant with other bacterial infections. The remainder were A. pleuropneumoniae, unclassified Actinobacillus species and Pasteurella caballi. Actinobacillus equuli including subsp. haemolyticus and subsp. equuli were the species most frequently isolated from equine actinobacillosis in Japan.
Cancer consists of heterogeneous cells that contain a small population of cells that possess stem cell properties; these cells, referred to as cancer stem cells (CSCs) or tumor-initiating cells, are involved in tumor progression and metastasis. Using a sphere-forming assay, canine mammary CSCs were found to be similar to human breast CSCs. Metabolic reprogramming has been recognized as a hallmark of various cancers. However, the significance of cellular metabolism in CSCs remains unclear. The aim of this study was to define the metabolic characteristics of CSCs derived from canine mammary tumors and gain an understanding of the maintenance of stemness. We identified metabolite profiles of canine mammary adenocarcinoma cell lines using gas chromatography-mass spectrometry. Metabolites were extracted from both adherent and sphere-forming cells derived from three cell lines. Sphere-forming cells were separated from adherent cells using an orthogonal, partial least-squares discriminant analysis. Sphere-forming cells were found to contain high levels of the amino acids alanine, glycine and proline compared with adherent cells. They also had high levels of palmitoleate, palmitate and dihomo-gamma-linolenic acid compared with adherent cells. In a sphere-forming assay, palmitate increased the number of spheres for all cell lines. These results indicate that the sphere-forming cells derived from canine mammary adenocarcinoma cell lines have specific metabolic profiles that may be useful for the development of CSC-specific therapies targeting metabolic pathways and potential stemness biomarkers; these results also clarify the maintenance of stemness in canine mammary CSCs.
Inflammatory bowel disease (IBD) is a common gastrointestinal disease in dogs. Decreased production of intestinal immunoglobulin A (IgA) has been suggested as a possible pathogenesis in a subset of canine IBD; however, the underlying cause remains unclear. Sphingosine-1-phosphate (S1P) is a lipid mediator that regulates intestinal IgA production by controlling lymphocyte trafficking in mice. The objectives of this study were to clarify the role of S1P in IgA production in dogs and to evaluate the expression of S1P-related molecules in dogs with IBD. First, an S1P receptor antagonist was administrated to five healthy dogs. The S1P receptor antagonist significantly decreased the IgA concentration in sera and feces but did not affect the IgG concentration. Moreover, the immunoreactivity of intestinal IgA was significantly decreased by S1P signal blockade. These results indicate that S1P signaling specifically regulates the intestinal IgA production in dogs. Subsequently, the intestinal S1P concentration and the expression of S1P-related molecules were measured in dogs with IBD and healthy dogs. The intestinal concentration of S1P was significantly lower in dogs with IBD than in healthy dogs. In addition, the gene expression levels of S1P receptor (S1P1) and S1P synthase (SK1) were significantly lower in dogs with IBD than in healthy dogs. Taken together, these observations suggest that decreased S1P production, likely caused by a lower expression of S1P synthetase, leads to attenuation of S1P/S1P1 signaling pathway and the production of intestinal IgA in dogs with IBD.
We monitored changes in serum leptin, adiponectin, and resistin concentrations in obese cats during weight loss. Six naturally developed obese cats were fed low-fat, high-fiber dry food during a 9-week experimental period. Serum leptin, adiponectin, and resistin concentrations were measured at week 0, 4, 8, and 9. Body weight became significantly lower week 4 onward than that at week 0 (P<0.05 or 0.01). At week 9, serum leptin concentrations were significantly lower than those at week 0 (P<0.05). Contrarily, serum adiponectin and resistin concentrations did not significantly differ within the 9 weeks. While serum leptin levels were strongly positively correlated with body weight (r=0.923, P<0.001), serum adiponectin levels were moderately negatively correlated with it (r=−0.529, P<0.01), with serum resistin having a no correlation with body weight. Serum leptin levels might be more closely related with pathogenesis of adiposity than serum adiponectin or resistin in cats.
A 12-year-old neutered female American cocker spaniel weighing 9.9 kg was presented for evaluation with a 2-day history of dyspnea and anorexia. Echocardiography revealed severe pulmonary hypertension (estimated systolic pulmonary arterial pressure, 93.4 mmHg) with right heart enlargement, pulmonary arterial dilation, and right ventricular dysfunction. The dilation of left heart and congenital cardiac shunt were not observed. Pulmonary thromboembolism (PTE) was confirmed by computed tomographic angiography. After treatment with antiplatelet and anticoagulant, the clinical sign and the echocardiographic abnormality of right heart were improved. These echocardiographic findings are not specific for PTE, but it can be useful as a rule-in test for PTE when other causes of pulmonary hypertension are excluded and a monitor of therapeutic efficacy.
The aim of this study was to evaluate the accuracy and precision of portable blood glucose meters, such as i-STAT 1 and Precision Xceed, for use in calves. Whole blood and plasma samples were obtained from eleven calves that received 2.5 or 5.0% dextrose-containing polyelectrolyte isotonic solutions. Measurements using the i-STAT 1 (r2=0.99, P<0.0001) and Precision Xceed (r2=0.96, P<0.0001) were well correlated with those by the hexokinase method, which is the gold standard. Although the accuracy of i-STAT 1 was equivalent to that of the hexokinase method, there was an autocorrelation in the residuals between the results from the Precision Xceed and the hexokinase method. Thus, the i-STAT 1 can be used to measure the blood glucose concentration in cattle.
To evaluate the preventive effects of combined vaccination for Pasteurella multocida, Mannheimia haemolytica and Histophilus somni on respiratory diseases in Japanese Black calves, 295 calves at one farm were alternately assigned to two groups; 147 calves received the vaccine at 4 and 8 weeks of age (vaccination group), and the other 148 calves did not receive vaccine (control group). The incidences of respiratory diseases were 25.9 and 70.9% in the vaccination and control groups, respectively, and the odds ratio for comparison between the two groups was 0.143 (95% confidence interval: 0.086–0.238). Administration of the multiple vaccine to Japanese black calves might be one of effective factor for prevention of respiratory diseases.
Baicalin is a plant-derived flavonoid that has anti-inflammatory and anti-oxidative effects. We investigated an anti-inflammatory effect of baicalin against lipopolysaccharide (LPS)-induced damage in cerebral cortex. Adult mice were divided into control, LPS-treated, and LPS and baicalin co-treated animals. LPS (250 µg/kg/day) and baicalin (10 mg/kg/day) were intraperitoneally injected for 7 days. LPS treatment induced histopathological changes in cerebral cortex, whereas baicalin protected neuronal cells against LPS toxicity. Moreover, baicalin treatment attenuated LPS-induced increases of reactive oxygen species and oxidative stress in cerebral cortices. Ionized calcium binding adaptor molecule-1 (Iba-1) and glial fibrillary acidic protein (GFAP) are known as markers of activated microglia and astrocyte, respectively. Results of Western blot and immunofluorescence staining showed that LPS exposure induces increases of Iba-1 and GFAP expressions, whereas baicalin alleviates LPS-induced increases of these proteins. Baicalin also prevented LPS-induced increase of nuclear factor kappa B (NF-κB). LPS treatment led to increases of pro-inflammatory factors including interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Increases of these pro-inflammatory mediators were attenuated in baicalin co-treated animals. These results demonstrated that baicalin regulates neuroglia activation and modulates inflammatory factors in LPS-induced neuronal injury. Thus, our findings suggest that baicalin exerts a neuroinflammatory effect against LPS-induced toxicity through decreasing oxidative stress and inhibiting NF-κB mediated inflammatory factors, such as IL-1β and TNF-α.
In a previous study, we showed that 2’, 3’-Cyclic Nucleotide 3’-Phosphodiesterase (CNPase) expression is induced in different temporal patterns in the cerebrum, cerebellum and medulla oblongata of hexachlorophene (HCP) and cuprizone (CPZ) treated rats. Here, we additionally examined the histopathological changes and CNPase expression in the spinal cord to clarify the reproducibility of different temporal patterns of CNPase expression in the spinal cord showing low degree or lack of spongy changes. Spongy changes were observed in HCP-treated rats, but not in CPZ-treated rats. Immunohistochemistry showed that intense expression of CNPase was not induced following HCP or CPZ treatment. Our data reveal that expression intensity of CNPase may be dependent on the degree of HCP- and CPZ-induced damage of the myelin sheath.
A one-year-old male Maltese terrier presented with mild ataxia and disorientation for 4 months. Over time, clinical signs progressed from paraparesis to non-ambulatory tetraparesis, voice change and dysphagia. Histological examination revealed concurrent leukoencephalomyelitis and polyneuritis. Infectious etiologies, including dengue, Japanese encephalitis, Zika, canine distemper, pseudorabies, rabies, toxoplasmosis, neosporosis, leishmaniasis, and encephalitozoonosis, were ruled out by PCR and/or immunohistochemical (IHC) staining. IHC tested on neurological tissues highlighted a heterogeneous population of infiltrating T and B lymphocytes admixed macrophages. Therefore, this case was diagnosed with current leukoencephalomyelitis and polyneuritis, resembling combined central and peripheral demyelination (CCPD), an autoimmune inflammatory demyelinating disease affecting both the CNS and PNS in humans.
The aim of the present study was to clarify roles of ATP-dependent potassium channels (KATP channels) in motility of the striated muscle portion in the esophagus. An isolated segment of the rat esophagus was placed in an organ bath and mechanical responses were recorded using a force transducer. Electrical stimulation of the vagus nerve evoked contractile response of striated muscle in the esophageal segment. Application of glibenclamide, an antagonist of KATP channels, increased amplitude of vagally mediated twitch contractions of the rat esophagus. On the other hand, minoxidil, an agonist of KATP channels, decreased amplitude of twitch contractions. RT-PCR revealed the expression of subunits of KATP channels in esophageal tissue. In addition, immunopositivity for subunits of KATP channels was observed in the striated muscle cells of the esophageal muscle layer. These findings indicate that KATP channels contribute to motor regulation of striated muscle in the rat esophagus.
Recently, it has been proposed that neuromedin U (NMU) is “decretin”, which suppresses insulin secretion from the pancreas in vitro. Here we examined the possible involvement of NMU in insulin secretion in vivo by comparing the plasma glucose and insulin levels of wild-type mice with those of double knockout (D-KO) of the NMU and neuromedin S (NMS) genes, as NMS binds to the neuromedin U receptor. If NMU is, in fact, “decretin”, which inhibits insulin secretion from the pancreas, then NMU-deficient mice might result in higher plasma insulin levels than is the case in wild-type mice, or injection of NMU lead to suppression of plasma insulin level. In this study, we found that the fasting plasma level of insulin was not increased in D-KO mice. Glucose tolerance tests revealed no significant difference in plasma insulin levels between wild-type mice and D-KO mice under non-fasting conditions. After peripheral injection of NMU, plasma glucose and insulin levels did not show any significant changes in either wild-type or D-KO mice. Glucose tolerance testing after 3 weeks of high fat feeding revealed no significant difference in plasma insulin levels during 60 min after glucose injection between wild-type and D-KO mice. These results suggest that even if NMU is a decretin candidate, its physiological involvement in suppression of insulin secretion may be very minor in vivo.
We investigated the association among endometrial hyperemia, uterine bacterial infection, and features of the large ovarian follicles in dairy cows. Genital organs were collected in a complete set at a slaughterhouse, and the degree of endometrial hyperemia was examined for the direct evaluation of uterine inflammation. The rate of bacterial infection in the uterus was higher in cows with endometrial hyperemia regardless of the severity of hyperemia, compared with cows without hyperemia. Moreover, the characteristics of the follicular fluid were changed in cows with uterine bacterial infection and included high concentrations of lipopolysaccharide and malondialdehyde (lipid peroxidation marker). These findings can be utilized as the basic information for the direct evaluation of the uterine inflammatory status in dairy cows.
Food additive grade calcium hydroxide (FdCa(OH)2) in the solution of 0.17% was evaluated for its bactericidal efficacies toward Legionella pneumophila with or without sodium hypochlorite (NaOCl) at a concentration of 200 ppm total residual chlorine, at room temperature (RT) (25°C ± 2°C) and 42°C, either with or without 5% fetal bovine serum (FBS). Besides, FdCa(OH)2 in different concentration solutions were prepared in field water samples (hot spring and bath tab water) and evaluated for their bactericidal efficacies at 42°C. FdCa(OH)2 (0.17%) inactivated the L. pneumophila to the undetectable level (<2.6 log CFU/ml) within 5 min and 3 min, respectively, at RT and 42°C, with 5% FBS. At RT and 42°C, NaOCl inactivated L. pneumophila to the undetectable level within 5 min, without 5% FBS, but with 5% FBS, it could only inactivate this bacterium effectively (≥3 log reductions). Conversely, at RT and 42°C, the mixture of 0.17% FdCa(OH)2 and 200 ppm NaOCl could inactivate L. pneumophila to the undetectable level, respectively, within 3 min and 1 min, even with 5% FBS, and it was elucidated that FdCa(OH)2 has a synergistic bactericidal effect together with NaOCl. FdCa(OH)2 0.05% solution prepared in hot spring water could inactivate L. pneumophila to the undetectable within 3 min at 42°C. So, FdCa(OH)2 alone could show nice bactericidal efficacy at 42°C, even with 5% FBS, as well as in field water samples.
During fiscal years 2014–2018, a total of 254 internship programs were held by local government organizations under the VPcamp project, a project sponsored by the Japanese Ministry of Education, Science and Technology. We conducted a Poisson regression analysis using the number of applicants for each program as dependent variable and potential factors as independent variables that might affect the number of applicants. The factors that were found to significantly affect the number of applicants were: the program opening date; the type of venue where the program took place; the target grades of veterinary students; the regional location of the local government that organized the program; and the proximity of the local government from a veterinary school.
This study describes the clinical presentation of superficial swellings and evaluates the utility of ultrasonography in the diagnosis of such swellings in sheep and goats. Ninety-three sheep and 73 goats were included in this study on the basis of presence of superficial swellings on the head (n=56), neck (n=16), chest wall (n=3), abdominal wall (n=40), umbilicus (n=14), scrotum (n=16), testes (n=1), udder (n=7), limbs (n=2), gluteal region (n=5), tail (n=1), and penile urethra (n=5). Ultrasonographic evaluation of these superficial swellings allowed the diagnosis of abscesses (n=54; 32.52%), cysts (n=12; 7.23%), hernias (n=57; 34.33%), hematomas (n=14; 8.44%), tumors (n=24; 14.45%), and urethral diverticula (n=5; 3.03%). Each lesion type could be precisely discriminated (sensitivity, 88–100%; specificity, 80–100%; and P=0.001). Ultrasonography was found to have a specificity of 100% for the diagnosis of hernias, urethral diverticula, and tumors, and a lower specificity of 80% for hematomas and 93% for abscesses when used for evaluation of superficial swellings in sheep and goats. In conclusion, ultrasonography is a unique, non-invasive diagnostic imaging tool that allows the diagnosis, differential diagnosis, and subsequent surgical treatment of different types of superficial swellings in sheep and goats.
In healthy dogs, amino acid infusion significantly attenuates the decrease in body temperature during anesthesia by facilitating insulin secretion, suggesting that such an increase in insulin secretion is related to increased heat production. In dogs, selective cyclooxygenase-2 (COX-2) inhibitors, which are used for pain relief in veterinary medicine, possess anti-pyretic action. And, in mice and humans, selective COX-2 inhibitors increase insulin secretion and sensitivity. Therefore, treatment with COX-2 inhibitors may negate or accelerate the attenuating effect on decreased body temperature during anesthesia by amino acid infusion. In the present study, influences on insulin secretion and body temperature by treatment with meloxicam or robenacoxib at therapeutic dose were evaluated in healthy dogs. Treatment with meloxicam or robenacoxib did not affect insulin secretion in the unanesthetized and anesthetized dogs, and did not affect body temperature and heart rate under the anesthetized condition with amino acid infusion. In conclusion, COX-2 inhibitors at therapeutic doses did not affect body temperature during anesthesia in dogs administered amino acids.
The objective of the present study was to investigate whether double intra-vulvo submucosal administration of prostaglandin F2α (PGF2α) could improve farrowing synchronization compared to single administration, and the effects of the induction of farrowing on colostrum yield and piglet performances. In total, 91 sows were randomly assigned to one of three groups: i) treatment with a single or ii) double administration of PGF2α or iii) control group without any treatment. A synthetic analogue of PGF2α was administrated via the intra-vulvo submucosal route on day 114 of gestation, at 0800 hr (single administration) or at 0800 and 1400 hr (double administration). The animals were monitored during the farrowing process. The time interval from the first administration of PGF2α to the onset of parturition was compared between groups. The proportion of sows that farrowed within 32 hr of induced parturition was higher in the double administration group than in the single administration group (100 vs. 84.4%, P=0.046). The duration of farrowing was higher in single administration sows than in control (241.1 vs. 169.5 min, P=0.004) and tended to be higher than in double administration sows (241.1 vs. 190.3 min, P=0.088). Birth interval of piglets born after double administration of PGF2α was shorter than those born after a single administration (14.6 and 20.1 min, P=0.024). In sows, the induction of parturition using a double administration of PGF2α reduces variation in gestation length and significantly increases the proportion of sows with an early response to PGF2α.
In this study, we aimed to elucidate the utility of transrectal ultrasonography (TRUS) and blood lactate concentration (bLac) measurement to diagnose cows with severe uterine torsion. We investigated the association of TRUS and bLac measurement with macroscopic findings on laparotomy for severe uterine torsion in nine cows. We found that an increased ultrasonographic cross-sectional thickness (15–25 mm) and multiple hypoechogenic areas corresponded to macroscopic vascular compromise in the uterus on laparotomy. In addition, bLac was elevated (≥5.0 mmol/l) in cows showing uterine necrosis on laparotomy. A combined diagnostic approach with TRUS and bLac measurement enables assessment of the uterine vascular status and has utility for selecting the treatment option (including laparotomy) and predicting the outcomes.
Following an outbreak of classical swine fever (CSF) in Japan, 2018, CSFV JPN/1/2018 was isolated from an infected pig sample. In this study, we carried out a comparative experimental infection in pigs using this strain and the highly virulent ALD strain and compared outcomes, including clinical manifestation, virus shedding patterns and antibody responses. Although pigs inoculated orally or intramuscularly with JPN/1/2018 developed hyperthermia and had decreased leucocyte numbers, they survived for the whole experimental period and showed less severe clinical signs than those infected with the ALD strain. We confirmed the presence of characteristic multifocal infarction of the margin of the spleen that arises following infection with JPN/1/2018, albeit that this finding was not observed in all infected pigs. Both viruses efficiently spread to contact pigs in a similar manner, suggesting in transmissibility between the two strains. Viral RNAs were detected in all clinical samples, especially whole blood samples, before the pigs developed hyperthermia until at least approximately 2 weeks after inoculation. Our findings will be valuable for the investigations into epidemic events occurring in Japan and for establishing diagnostic strategies and control measures against CSF.
In the testes of the Sunda porcupine (Hystrix javanica), the expression of the steroidogenic acute regulatory protein (StAR) and steroidogenic enzymes, such as cytochrome P450 side chain cleavage (P450scc), 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 17α-hydroxylase (P450c17) and cytochrome P450 aromatase (P450arom), was immunohistochemically examined to clarify the location of steroidogenesis. In this study, complete spermatogenesis (spermiogenesis) was observed in the testes of the examined Sunda porcupine, and spermatozoa of the Sunda porcupine had a spatulate sperm head unlike that of rats and mice which has an apical hook. On immunostaining of StAR, P450scc, 3β-HSD, P450c17 and P450arom, immunoreactivity for all proteins was only detected in the Leydig cells and not observed within the seminiferous tubules, suggesting that the Leydig cells can synthesize both androgen and estrogen from cholesterol in the Sunda porcupine testes.
Influenza virus is known to affect wild felids. To explore the prevalence of influenza viruses in these animal species, 196 archival sera from 5 felid species including Panthera tigris (N=147), Prionailurus viverrinus (N=35), Panthera leo (N=5), Pardofelis temminckii (N=8) and Neofelis nebulosa (N=1) collected between 2011 and 2015 in 10 provinces of Thailand were determined for the presence of antibody to avian and human influenza viruses. Blocking enzyme-linked immunosorbent (ELISA) assay and hemagglutination inhibition (HI) assay were employed as the screening tests, which the serum samples with HI antibody titers ≥20 were further confirmed by cytopathic effect/hemagglutination based-microneutralization (CPE/HA-based microNT) test. Based on HI and microNT assays, the seropositive rates of low pathogenic avian influenza (LPAI) H5 virus, highly pathogenic avian influenza (HPAI) H5 virus and human H1 virus were 1.53% (3/196), 2.04% (4/196) and 6.63% (13/196), respectively. In addition, we also found antibody against both LPAI H5 virus and HPAI H5 virus in 2 out of 196 tested sera (1.02%). Evidences of influenza virus infection were found in captive P. tigris in Kanchanaburi, Nakhon Sawan and Ratchaburi provinces of Thailand. The findings of our study highlights the need of a continuous active surveillance program of influenza viruses in wild felid species.
The transgenerational maintenance of symbiotic microbes that benefit host nutrition and health is evolutionarily advantageous. In some vertebrate lineages, coprophagy is used as a strategy for effectively transmitting microbes across generations. However, this strategy has still not been studied in birds. Accordingly, the aim of the present study was to evaluate the role of maternal cecal feces consumption by Japanese rock ptarmigan (Lagopus muta japonica) chicks as a strategy for acquiring essential gut microbes. Both the duration of coprophagy behavior by the chicks and the development process of the chick cecal microbiome (n=20 one- to three-week-old chicks, from three broods) were investigated. In all three broods, coprophagy behavior was only observed from 3 to 18 days of age. Furthermore, there was no significant difference in the number of bacterial operational taxonomic units (OTUs) in 1-week-old chicks (n=651) and adults (n=609), and most of the main OTUs observed in the adults were already present in the 1-week-old chicks. These results indicate that, in this precocial bird species, coprophagy may contribute to the early establishment of cecal bacteria that are essential for food digestion and, thus, chick survival. In fact, Japanese rock ptarmigan chicks consume the same food as their hens from the time of hatching. This behavior may have applications to ex-situ conservation.
A wild adult Eurasian scops owl (Otus scops), which was unable to fly, was rescued. Physical examination revealed a sticky exudate around the glottis. Heterophilic leukocytosis was identified through complete blood count, and radiography revealed a marked elevated density of posterior air sacs and inner cavities in both sides of the humerus and femur. Fungal cultures of samples taken from the owl suggested a respiratory fungal infection. Through molecular typing, the fungus was identified as Epicoccum nigrum. The owl was treated with oral itraconazole and broad-spectrum antibiotics. After one month, the inner cavities of pneumatic bones were slightly distinguishable by radiography and the owl started to fly well. Two months later, the air sac and all pneumatic bones displayed normal appearance.