Serum amino acid profiles in clinically normal Noma horses.

Plasma or serum amino acids are used to evaluate nutritional status and metabolic disorders. In this study, we aimed to set reference values of serum amino acid concentrations in the Noma horse, a Japanese native horse. Thirty-one horses were classified into six age groups: neonatal foal (0-4 days), foal (0.5-1 years), youth (5 years), middle age (10 years), old (15 years), and extra-old (>20 years). Horses >5 years of age were analyzed together as the adult group. In the adult horses, there were no significant differences among the serum amino acid concentrations of each age group. The foal group had higher concentrations of alanine, aspartic acid, glutamic acid, α-aminoadipic acid, and 3-methyl-histidine than the adult group. The neonatal foal group had higher serum concentrations of phenylalanine, lysine, alanine, proline, aspartic acid, glutamic acid, ß-alanine, and ß-amino-iso-butyric acid and lower tryptophan concentrations and Fischer's ratios than the adult group. The neonatal foal group had higher ß-amino-iso-butyric acid concentrations and lower tryptophan and 3-methyl-histidine concentrations than the foal group. Therefore, reference values might be set separately in neonatal foals, foals, and adult horses. The data for the serum amino acid concentrations can be used for health care through physiological and pathological evaluations in Noma horses.

Pathological findings of mammary gland carcinoma and hemangiosarcoma cases in two African wild dogs (Lycaon pictus).

There are few studies on diseases affecting endangered African wild dogs. We report our findings on malignant tumors in two African wild dogs. Case 1 was a 6-year-old intact female diagnosed with inflammatory mammary carcinoma with pulmonary metastasis. Case 2 was an 11-year-old male diagnosed with primary hemangiosarcoma of the left atrial coronary sulcus with metastasis to multiple organs. Additionally, the tumor had grown through the cardiac wall, causing cardiac tamponade. The identification of disease incidence trends provides important information which will allow for the early detection and treatment of malignant tumors, and aid in the conservation of this species.

Postmortem magnetic resonance imaging findings of tricuspid atresia with ventricular and atrial septal defects and subvalvular pulmonic stenosis in a Japanese native Noma horse.

The necropsy of a 2-day-old Noma horse that died of weakness showed an enlarged cardiac base and a narrow cardiac apex, suggesting cardiac malformation. The excised heart underwent imaging to investigate its luminal structure. On three-dimensional magnetic resonance imaging, the right atrium and right ventricle were discontinuous. The right atrium communicated with the left atrium and the left ventricle communicated with the right ventricle. The lumen narrowed near the pulmonary artery valve. Since the same findings were observed on gross examination, the foal was diagnosed with tricuspid atresia with ventricular and atrial septal defects, along with subvalvular pulmonic stenosis.

Spontaneous multiple cutaneous mixed tumors in Japanese giant salamander Andrias japonicus.

We examined 7 cutaneous mixed tumors in 2 wild-captured Japanese giant salamanders Andrias japonicus. The tumors were either already present and/or increased in size, or newly occurred during capativity. We sampled the 7 tumors from these animals and 3 verrucose protrusions from 3 unaffected animals, as controls, and examined them pathologically and virologically. The tumors (5 mm to 4 cm in size) were papillary protrusions or pendulated on the skin surface. The cut surface of the tumors was white, lobulated, partially hard, and contained mucus. All tumors presented similar histological characteristics of a hyaline structure and exhibited biphasic proliferation, with neoplastic epithelial cells partially composing the pseudo-ductal structure and staining positive for cytokeratin AE1/AE3. Vimentin 3B4-positive blast-like mesenchymal cells proliferated to fill the gaps in the epithelial components. Transition from unique mucous gland to neoplastic tissue was observed. The hyaline structure was stained blue by AZAN stain, Alcian blue-periodic acid-Schiff (PAS) double stain, and toluidine blue (TB) stain of pH 7.0, but was unstained by TB with pH values of 4.1 and 2.5. The mucus in the neoplastic tissue and in the mucous gland in verrucose protrusions was stained blue by Alcian blue-PAS double stain; TB staining at pH 7.0, 4.1, and 2.5 revealed metachromasy. No virus was detected in the tumors. The 7 tumors were diagnosed as cutaneous mixed tumors, and it was confirmed that the neoplastic cells originated from the mucous gland in the dermis. The biological behavior and pathological development of tumors should be elucidated because the tumors have the potential to negatively affect A. japonicus.

First demonstration of Strongyloides parasite from an imported pet meerkat - Possibly a novel species in the stercoralis/procyonis group.

Strongyloides is a genus of parasitic nematodes of vertebrates that contains over 50 species, each with a variable host range. A recent molecular phylogenetic analysis on this genus showed that Strongyloides spp. from various carnivore hosts form a strongly supported clade together with Strongyloides stercoralis, a major pathogen of humans and dogs (named the "stercoralis/procyonis group"). In the present study, we obtained DNA sequencing data of Strongyloides sp. isolated from an imported meerkat (Suricata suricatta). Based on the phylogenetic analysis, we considered this a new member of the stercoralis/procyonis group. This study represents the first isolation and molecular characterization of a Strongyloides species from hosts belonging to the family Herpestidae (mongooses and meerkat). However, whether the meerkat serves as a natural host of this Strongyloides species remains to be investigated.

First report of ophidiomycosis in Asia caused by Ophidiomyces ophiodiicola in captive snakes in Japan.

Ophidiomycosis is an emerging infectious disease caused by the fungus Ophidiomyces ophiodiicola, which has been affecting wild and captive snakes in North America, Europe, and Australia. We report 12 cases of suspected ophidiomycosis in captive colubrid snakes in Japan. Pathological and microbiological examinations were performed, and the results confirmed the diagnosis of ophidiomycosis in two snakes, which indicated that the remaining sympatrically raised snakes also had ophidiomycosis since they exhibited similar lesions. This is the first report of ophidiomycosis in Asia caused by O. ophiodiicola. To prevent the expansion of ophidiomycosis in the natural environment in Japan, there is a need to evaluate the ophidiomycosis carrier status of imported snakes, the pathogenicity of the infection in native snakes, and the prevalence and distribution of O. ophiodiicola in wild and captive snakes. Measures also must be taken to prevent endemicity globally.

Rabbit hemorrhagic disease virus type 2 epidemic in a rabbit colony in Japan.

Twenty-three of 42 European rabbits (Oryctolagus cuniculus), belonging to the same rabbit colony, died in March 2020 (55% mortality) in Chiba prefecture, Japan. The disease course was extremely acute without indicators of death or hemorrhage. Necropsy revealed liver swelling, discoloration, cloudiness and fragility, and pulmonary edema. Histologically, severe hepatocellular necrosis (mainly peripheral) and intra-glomerular capillary hyalin thrombi were observed. On molecular-biological examination, reverse transcription polymerase chain reaction analysis of RNA from tissues detected a rabbit hemorrhagic disease virus, confirmed as a RHDV-2 VP60 fragment, which shared 99.42% nucleotide identity with the homologous fragment of RHDV-2 German isolate by nucleotide sequence analysis. This report shows the outbreak of rabbit hemorrhagic disease caused by RHDV-2, an emerging infectious disease, in Japan.

Pyogenic meningitis and trigeminal neuritis secondary to periodontogenic paranasal sinusitis in a red deer (Cervus elaphus).

An adult female red deer died of a severe seizure and dysbasia. Postmortem computed tomography (CT) and magnetic resonance imaging (MRI) were performed. On CT, deciduous right maxillary second and third premolar teeth were observed, and the right infraorbital canal was disrupted. MRI showed that the right trigeminal nerve was enlarged and the right subarachnoid cavity was occupied by fluid and gas. On gross examination, the right paranasal sinus, swollen muscles of the orbit and tonsils, right trigeminal nerve, and right cerebrum surface contained a yellowish-white, cheese-like pus. Based on these findings, the deer was believed to have developed pyogenic meningitis caused by a neuropathic infection secondary to periodontogenic paranasal sinusitis.

New canine parvovirus 2a infection in an imported Asian small-clawed otter (Aonyx cinereus) in Japan.

Post-import from the Republic of Indonesia to Japan in 2017, two juvenile, captive bred Asian small-clawed otters (Aonyx cinereus) exhibited gastrointestinal symptoms, including vomiting, diarrhea, and hematemesis, and died. One of them was examined postmortem. Microscopically, the small intestinal mucosa was necrotic with crypts lined by regenerating large epithelial cells. A gastric cardiac mucosal ulcerative lesion containing fungal yeasts and pseudohyphae morphologically indicated Candida spp. The lymph nodes exhibited marked lymphoid depletion. Canine parvovirus 2 (CPV-2) was isolated from an oral swab, and virus protein 2 (VP2) gene sequencing revealed new CPV-2a. To our knowledge, this is the first new CPV-2a infection report in Asian small-clawed otters. This infection should be considered in gastrointestinal symptom-related cases in this species.

Comparison of methods for detection of chytrid fungus (Batrachochytrium dendrobatidis) in bullfrog tadpole mouthparts.

We previously reported that the tadpole of bullfrog (Lithobates catesbeiana) is a useful model for the field surveillance of the Batrachochytrium dendrobatidis (Bd) distribution. In the present study, we compared Bd detection rates in swab-scraped and resected mouthpart samples, using nested polymerase chain reaction (PCR). The resulting detection rates for swab-scraped and resected specimens were 67 and 65%, respectively, with no significant difference. Furthermore, we performed a histopathological examination for Bd distribution in the mouthparts; we found that Bd infection occurred in the tip and basement of the jaw sheaths and tooth rows. We recommend using swab-scraped samples for Bd detection. Moreover, careful attention should be paid to scraping the tip and basement of the jaw sheaths and the entire oral cavity to reduce the rates of false-negative results on nested PCR of the mouthparts of bullfrog tadpoles.

Spontaneous oral chytridiomycosis in wild bullfrog tadpoles in Japan.

Batrachochytrium dendrobatidis (Bd) infects Anuran larvae (tadpole) mouthparts and causes oral chytridiomycosis, which can be diagnosed in tadpoles by detecting mouthparts deformities. However, oral chytridiomycosis may or may not be observable, depending on species, tadpole stage and season, and has never been reported in Japan. We aimed to observe oral chytridiomycosis characteristics in bullfrog (Lithobates catesbeiana) tadpoles, determine associated pathologic features and investigate the usability of bullfrog tadpoles in Japanese Bd field surveys. Wild-captured bullfrog tadpole mouthparts were examined macroscopically, histopathologically and by molecular biological examination. Macroscopic lesions were observed in 21 of 59 tadpole mouthparts. Lesions were most frequently located in the lower jaw sheaths and were mainly recognized by partial depigmentation (11 tadpoles; some were completely depigmented) and thinning of the pigmented layer (10 tadpoles). Partial defects of the tips and blunt cutting edges of the jaw sheaths were observed with severe jaw sheath depigmentation. Whitened tooth rows were observed in 7 tadpoles. Histologically, the stratified epithelium (pigmented epithelium) showed partial or diffuse hypopigmentation or pigment loss. Irregular stratified epithelium thickening with hyperkeratosis or parakeratosis was observed in the jaw sheaths. Bd infection was confirmed in 20 of 21 tadpoles presenting jaw sheath deformities, by histopathological examination and/or nested polymerase chain reaction. Depigmentation and thinning of the pigmented layers of jaw sheaths were associated with Bd infection. Thus, diagnosis of Bd infection by macroscopic observation of bullfrog tadpole mouthparts is feasible. This is the first report of oral chytridiomycosis in wild bullfrog tadpoles in Japan.

Oxalate nephropathy in free-living American bullfrog tadpoles.

In February 2014, wild American bullfrog Lithobates catesbeianus tadpoles from an artificial pond in the Kyusyu region, Japan, presented with coelomic and subcutaneous edema and erythema within the skin. A pathological examination of 57 tadpoles of American bullfrogs in the region was conducted to evaluate the disease. Crystal deposition of varying degrees was found in the kidneys of 35 tadpoles (61.4%). The crystals were transparent, pleomorphic in shape, highly birefringent in polarized light, and arranged in a radial pattern within the renal tubular lumen. Using Alizarin Red S stain and liquid chromatography, these crystals were identified as calcium oxalate. Severe coelomic and subcutaneous edema was observed in 7 of these 35 tadpoles (20.0%). Ammonia levels in coelomic fluid were extremely elevated (>1000 µg dl(-1)) in 4 tadpoles examined. These findings suggest that oxalate deposition in kidneys causes metabolic disorder with renal nephropathy. The source of the oxalate could not be determined; however, the presence of calcium oxalates in pond sediments, as revealed by liquid chromatography, suggested that the deposition was most likely due to ingestion of oxalate materials from the environment. This is the first report of oxalate nephropathy in free-living amphibians.

First report of spontaneous chytridiomycosis in frogs in Asia.

This is the first report of amphibian chytridiomycosis in Asia. We discovered a lethal outbreak in Japan, among 45 exotic frogs from 18 species kept for breeding by a private owner. Of these 45 frogs, 16 died and another 7 were found to be infected by chytrid fungus Batrachochytrium dendrobatidis (Bd) but survived after treatment. Bd was detected in frogs from 9 species (Lepidobatrachus laevis, Ceratophrys cornuta, C. cranwelli, C. ornata, C. calcarata, Chacophrys pierotti, Occidozyga lima, Leptodactylus pentadactylus and Plethodontohyla tuberata).