We examined the orbicularis oculi (OC) muscle and its innervation among the markedly modified facial muscles of odontocetes, clarifying features related to secondary adaptations to aquatic life. The OC muscle has been described as a well-developed sphincter encircling the eye in cetaceans; however, this study revealed two distinct types of OC morphology among odontocetes. In specimens from the families Kogiidae and Ziphiidae, the muscle bundles of the OC completely encircled the palpebral fissure in concentric circles, similar to that of Bovidae specimens. However, in specimens from Phocoenidae and Delphinidae, the OC featured a muscular area with muscle fibers and an aponeurotic area comprising collagen fibers lacking facial nerve distribution. Our results suggest that this aponeurotic area was acquired in the common ancestor of species within the Delphinoidea superfamily, and that the upper eyelid is less mobile than the lower eyelid. The palpebral branches, which originate from the facial nerve, extend from the lower to the upper palpebral regions in the examined odontocetes, showing a clear difference from the entry course seen at the lateral canthus in artiodactyls. The proximal palpebral branches diverged directly from the facial nerve trunk, forming a characteristic diverging point distal to that of artiodactyls. These branches probably correspond to the auriculopalpebral nerve, which has lost the rostral auricular branch component. The morphological changes in cetacean heads also led to changes in the branching pattern and pathway of the facial nerve.

Forty male brown bear heads were non-destructively analyzed by computed tomography. Skull, frontal, and cranial cavity volumes were measured, and growth curves and regression lines were constructed. The morphological dynamics of the frontal sinus were also examined. Skull volume increased rapidly from 0 to 11 years of age, then grew more slowly until 21 years, when it plateaued. The frontal sinus also increased rapidly from 0 years, reaching approximately 300 mL by 10 years, after which growth ceased. The frontal sinus-to-skull ratio rose quickly from 0 to 4 years, then increased more gradually until 10 years, and then plateaued. Morphologically, the frontal sinus developed posteriorly to cover the anterior part of the cranial cavity, extended ventrally in the anterior lateral part of the cranial cavity and the orbital region, and protruded into the zygomatic process of the frontal bone. The cranial cavity increased slowly from 0 years, with continued growth beyond 11 years at a reduced rate. The cranial cavity-to-skull ratio decreased because the skull volume significantly increased. Both the frontal sinus and cranial cavity volumes strongly correlated with skull volume, and a well-fitting regression line was obtained. This means that the volumes of the frontal sinus and cranial cavity correlate more closely with skull volume than with age. This is the first report on the non-destructive analysis of the changes in volume and morphology of the frontal sinus and cranial cavity in male brown bears.

Mucosa-associated lymphoid tissue, which plays a crucial role in both immunological protection and inflammation at mucosal sites, shows interspecies variation in its distribution. In the ocular region, lacrimal duct-associated lymphoid tissue (LDALT) has rarely been investigated in animals other than humans or rodents. Although conjunctiva-associated lymphoid tissue (CALT) has been reported in various species, its cell composition remains unclear. Herein, we aimed to elucidate the morphology and development of the feline ocular immune system by examining the lacrimal tract and its associated tissues in adolescent (7- to 8-month-old) and fetal cats. The feline lacrimal tract, composed of the lacrimal canaliculi, lacrimal sac, and nasolacrimal duct, was visualized using CT. Histologically, a small lymphocyte aggregate, identified as LDALT, was observed exclusively along the lacrimal canaliculus in one of the four adolescent cats. In contrast, extensive development of the CALT was observed in tissues surrounding the openings of the lacrimal tract, particularly on the cornea-facing side of the third eyelid. CALT is characterized by CD20⁺ B cell aggregates surrounded by CD3⁺ T cells, including a small subset coexpressing CD20 and CD3. Neither LDALT nor CALT was observed in late gestational fetuses. In conclusion, CALT appears to contribute more significantly than LDALT to feline ocular immunity. Furthermore, given the pro-inflammatory nature of both CALT and CD3⁺CD20⁺ cells, our findings suggest that CALT is involved in the pathology of feline conjunctivitis.

Arterial distribution to the tensor fasciae antebrachii (TFA) muscle is little described in the veterinary literature. In this study, we investigated TFA vascularity intra- and inter-specifically, using specimens obtained from dogs, cattle, pigs, and horses at necropsy. Canine TFA specimens showed vascularity from the collateral ulnar artery in all cases, and from the thoracodorsal artery in some cases, with no brachial or deep brachial arterial vascularity. Bovine TFA specimens showed vascularity from the thoracodorsal artery. All porcine TFA specimens showed deep brachial arterial vascularity. Equine TFA specimens showed strong subscapular arterial vascularity, as well as some supply from branches of the deep brachial artery. This article will encourage further active research.

Sialocele recurrence in dogs following surgical excision of the monostomatic sublingual gland is often attributed to incomplete removal of sublingual gland tissue posterior to the site of duct rupture, suggested as solitary lobules of the monostomatic sublingual gland. Here we examined these solitary lobules in six healthy beagle cadavers. Gross anatomical observation revealed solitary lobules distributed along the major sublingual duct and separated from the rostral lobe of the monostomatic sublingual gland. Histologically, the solitary lobules were connected to the major sublingual duct and showed a mixed glandular structure similar to the monostomatic sublingual gland but distinct from the polystomatic sublingual gland. These findings elucidate the anatomical basis of sialocele recurrence and emphasize the importance of completely excising solitary lobules.

This study investigated the relationship between herd average annual somatic cell count (Herd SCC) and epidemiological indicators of intramammary infection (IMI) in the nonlactating period. We also aimed to establish a predictive model and benchmark based on epidemiological indicators of non-lactating IMI (IMI indicators) for a dairy herd in Hokkaido, Japan. Individual SCC data were extracted from 3 years of dairy herd improvement (DHI) data from 3,173 dairy farms in Hokkaido, and three IMI indicators were calculated: first-calving cow infection rate (FCIR) from first-time DHI results, dry cow cure rate (DCR), and dry cow new infection rate (DNIR) based on pre- and postpartum DHI results. A SCC of ≥200,000 cells/mL in individual cows was defined as IMI, and IMI indicators were then calculated. Dairy farms were classified into five levels based on the Herd SCC shown in the DHI data. Significant correlations were observed between Herd SCC and all IMI indicators ( ρ =0.37–0.44). The correlations suggested significant positive relationships between Herd SCC, and average FCIR and average DNIR, respectively, and a significant negative relationship between Herd SCC and average DCR. Generalized linear model analysis confirmed the significance of the correlation between Herd SCC and IMI indicators, even after adjusting for environmental and management factors, such as housing system (P<0.01). By comparing the benchmarks obtained in this study, dairy farmers can achieve IMI control through appropriate management in the non-lactating period based on target values for improvement, depending on Herd SCC.

C-telopeptide (CTX) fragments of type I collagen are widely used as markers of bone resorption in mammals. This study evaluated the performance of two enzyme-linked immunosorbent assays (ELISAs) using antibodies against different CTX epitopes—CTP-18 and CTP-8—in ovariectomized (OVX) mice, and examined the cross-species applicability of the CTP-18 ELISA in Pacific bluefin tuna (Thunnus orientalis). In OVX mice, both serum and urinary CTP-18 levels significantly increased compared to sham-operated controls (P<0.05), and were moderately correlated with deoxypyridinoline (DPD) levels and inversely with bone mineral density. In contrast, CTP-8 was less responsive. The CTP-18 antibody cross-reacted with mouse, rat, human, and tuna sera, and showed good dilution linearity in tuna samples. Tuna serum CTP-18 levels were positively correlated with DPD (r=0.82, P<0.05). These findings suggest that the CTP-18-based ELISA is a reliable tool for monitoring bone resorption not only in rodents but also in fish, providing a new approach for assessing skeletal turnover in aquaculture species.

The mechanism of multi-azole antifungal resistance in the dermatophyte Microsporum canis was investigated through a transcriptome analysis of the multi-azole–resistant strain LLP25-013. Strain LLP25-013 was cultured with and without itraconazole (ITCZ), after which total RNA was extracted from mycelial samples. Whole-RNA sequencing was performed using a DNBSEQ-G400 system. A total of 8,846 expressed genes were detected in samples of both total RNAs from both ITCZ-treated and ITCZ-free samples. Genes sequenced with more than 10,000 reads were selected from ITCZ-treated and ITCZ-free samples, and the expression levels were compared. Treatment with ITCZ upregulated the expression of 63 genes compared with no ITCZ treatment. By contrast, ITCZ treatment downregulated the expression of 253 identified genes. ITCZ addition reduced the expression of many genes in M. canis, which could be related to the broad-spectrum antifungal drug resistance of this organism. Among the upregulated genes, the ATP-binding cassette (ABC) transporter gene is involved in resistance in other dermatophytes. Co-culture of strain LLP25-013 with milbemycin and other azoles suggested that the isolate acquired azole resistance through high expression of the ABC transporter.

Bovine mastitis may cause intractable, chronic infection in cases where the causative pathogen forms a biofilm; however, data are lacking on Staphylococcus aureus (S. aureus) biofilm in Japanese dairy herds, and the relative efficacy of antimicrobial agents to treat it. Accordingly, we aimed to investigate the biofilm-forming potential of S. aureus isolates obtained from the milk of Holstein dairy cows with clinical mastitis, and to elucidate the effects of five clinically used antibiotic drugs against S. aureus biofilms. We found that 14/24 S. aureus isolates formed biofilm. All five antibiotic drugs demonstrated an inhibitory effect on S. aureus at the planktonic stage (based on minimum inhibitory concentrations); however, only the three fluoroquinolone antibiotics (orbifloxacin, enrofloxacin, and marbofloxacin) and cefazolin inhibited biofilm formation (based on minimum biofilm inhibitory concentrations). However, none of the five antibiotics proved effective against formed biofilms (based on minimum biofilm-eradicating concentrations). Accordingly, we suggest that systemic fluoroquinolone antibiotic or intra-mammary cefazolin may prevent biofilm formation in mastitic cows infected with S. aureus, if administered at a sufficiently high concentration and a sufficiently early stage of the disease (before biofilms are actually formed), but that antibiotics may have little efficacy once the biofilm is established.

Canine epitheliotropic cutaneous T-cell lymphoma (ECTCL) is a malignant neoplasm exhibiting various skin lesions. Metastasis to the lymph nodes and distant organs contributes to the poor prognosis of ECTCL; however, the underlying molecular mechanisms remain unclear. In the present study, the roles of chemokine receptors, such as CC chemokine receptor (CCR) 4 and CCR7, in the migration of tumor cells were examined using a canine cutaneous lymphoma model. Three mouse groups were prepared and xenografted with wild-type (WT), CCR4 knockout (KO), or CCR7KO canine ECTCL (EO-1) cells. The proportion of EO-1 cells in tissues and blood was significantly lower in the CCR4KO and CCR7KO groups than in the WT group. Only the iliac lymph node, a sentinel lymph node for the xenograft site, was enlarged with the infiltration of EO-1 cells in all groups. The size of the iliac lymph node was smaller in the KO groups than in the WT group. All mice developed a subcutaneous nodule at the xenograft site, which was smaller in the KO groups. In the cell proliferation assay, chemokine ligands stimulated an increase in EO-1 cells in the WT group, but not in the KO groups. These results suggest that CCR4 or CCR7 plays a vital role in the initial migration to sentinel lymph nodes and proliferation in a canine cutaneous lymphoma model.

This report describes two dogs with high-grade multicentric B-cell lymphoma that relapsed 3 months after completing a 6-month-long chemotherapy regimen with L-asparaginase, cyclophosphamide, doxorubicin, vincristine, and prednisolone (L-CHOP). Both were re-treated with L-CHOP, achieving complete remission (CR) before undergoing sequential half-body irradiation (HBI; 6 Gy per fraction to cranial and caudal body, 2 weeks apart). Caudal HBI caused grade 3 gastrointestinal toxicity in both dogs and grade 3 thrombocytopenia in one; however, both remain in CR at the time of writing this report (i.e., 483 and 490 days post-relapse). These findings suggest that combining HBI with chemotherapy may offer benefits in relapsed canine lymphoma, especially when dogs are in CR at the time of HBI.

This study examined the effect of platelet-rich plasma (PRP) administration on the repair of articular cartilage damage in domestic pigs with experimentally induced leg weakness. The left hind stifle joints of six 56-day-old castrated domestic pigs were artificially injured; PRP was injected into the stifle joints of three pigs. Histopathological analysis of dissected articular cartilage revealed that all specimens were lightly stained with Alcian blue, and regions containing immature chondrocytes or chondrocyte clusters indicated cartilage repair. The average cartilage thickness was 189.44 μm in the control group and 386.53 μm in the treatment group, showing a significant difference (P≤0.028). These findings suggest that PRP administration promotes articular cartilage repair in domestic pigs.

Canine lymphoma is a common hematopoietic malignancy with limited therapeutic options, particularly in drug-resistant cases. Overexpression of Exportin 1 (XPO1) promotes oncogenesis by impairing the nuclear-cytoplasmic translocation of tumor suppressor proteins (TSPs). KPT-335 (Verdinexor), a selective XPO1 inhibitor, restores TSP function, inducing cell cycle arrest and apoptosis. In this study, we investigated the in vitro effects of KPT-335 alone and in combination with doxorubicin or vincristine in four canine lymphoma cell lines. KPT-335 reduced XPO1 protein expression in a dose-dependent manner, an effect reversed by proteasome inhibition, suggesting proteasome-mediated degradation. Combination treatments significantly suppressed cell proliferation compared with single agents. These findings highlight the preclinical evidence of combining KPT-335 with conventional chemotherapies in canine lymphoma.

This longitudinal study evaluated whether an introductory laboratory animal science lecture changes first-year veterinary students’ compassion toward laboratory animals. A pre- and post-lecture survey revealed a significant increase in total compassion scores. The educational impact varied based on students’ backgrounds; for instance, significant score increases were observed in students with prior pet ownership and those without experience in animal protection activities. Improvements were notable in attitudes toward allowing natural behaviors and the importance of legal regulations. A slight decrease in some anti-compassion item scores was also observed, potentially indicating the early formation of a professional utilitarian perspective. These findings suggest that foundational education is effective in enhancing compassion, suggesting that science-based knowledge is crucial for developing the professional ethical framework essential for a “culture of care”.

Acceptable euthanasia methods must rapidly induce unconsciousness and death without pain or distress. We evaluated intraperitoneal thiamylal sodium (TM; 150 mg/kg; hereafter TM150) in male Sprague–Dawley, Wistar, and Fischer 344 rats. TM produced immobilization, loss of righting reflex, respiratory arrest, and cardiac arrest in all strains, with the shortest latencies in Fischer 344. Plasma chemistry showed strain-dependent shifts, including aspartate aminotransferase (AST) increases and variable alanine aminotransferase (ALT) responses; chloride and calcium showed small, non-uniform changes across strains, and most values remained within commonly cited reference ranges. These findings most likely reflect perimortem physiology and methodological factors superimposed on strain baselines rather than primary organ injury. TM150 is therefore effective for rat euthanasia, although post-TM plasma biochemistry should be interpreted cautiously.

Isoleucyl-tRNA synthetase 1 (IARS1) is essential for protein synthesis. Although IARS1 mutations cause growth and metabolic disorders in humans, its role in embryonic development remains unclear. We generated Iars1 knockout (KO) mice and observed complete absence of homozygous KOs among 44 pups, indicating embryonic lethality. A χ² test confirmed significant deviation from Mendelian ratios. At E12.5, multiple resorbed implantation sites were observed, and histology at E9.5 showed embryos lacking recognizable structures. Evidence of implantation (decidual swellings with residual trophoblast and maternal vasculature) indicates post-implantation survival but failure to progress beyond early organogenesis. These results demonstrate that IARS1 is indispensable for early mouse development and provide a useful model to study the link between IARS1 deficiency and disrupted organogenesis.

Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity lung disorder caused by chronic exposure to Aspergillus fumigatus (Af). Existing mouse models do not fully capture the severity of ABPA. This study developed a novel murine model of ABPA using the spontaneously eosinophilic Yama mouse. Female Yama and BALB/c mice were sensitized and challenged with Asp f1, a major allergen derived from Af. Peripheral eosinophil counts, serum IgE levels, and histopathological changes in the lungs, including goblet cell hyperplasia, mucus plugging, collagen deposition, eosinophil cationic protein (ECP), and CD117/c-kit expression, were assessed. Cytokine and fibrosis-related gene mRNA expression of interleukin-4 (IL-4), IL-5, IL-6, IL-13, transforming growth factor-β (TGF-β), and Col1a1 was also measured. Compared to BALB/c mice, Asp f1-challenged Yama mice exhibited sustained eosinophilia, increased IgE, pronounced goblet cell hyperplasia with mucus plugging, collagen deposition, and elevated eosinophil cationic protein (ECP) expression. Yama mice also showed more significant peribronchial inflammation and pulmonary fibrosis. RT-PCR analysis demonstrated substantial increases in IL-4, IL-5, IL-6, IL-13, and Col1a1 transcripts, indicating a Th2-dominant immune profile and enhanced fibrotic activity. The minimal differences in TGF-β expression between Asp f1– and saline-treated mice indicate that fibrosis in Yama mice may proceed via a TGF-β-independent pathway. These findings establish the Yama mouse as a novel model for ABPA research, closely replicating the immunopathological features of severe human ABPA. This model provides a reliable platform to investigate eosinophil-driven lung fibrosis mechanisms and evaluate hypersensitivity responses contributing to human ABPA onset and progression.

Gastrointestinal (GI) tumors are common neoplastic diseases in dogs. However, epidemiological data on canine GI tumors in Japan are limited. The present study aimed to investigate the prevalence of GI tumors in Japan and assess the association of canine breed, age, sex, and anatomical location with the development of common tumor types. A total of 1,310 canine GI tumors that were histopathologically examined between 2012 and 2024 were retrospectively analyzed. The statistical methods included a contingency table analysis, multivariable logistic regression analyses, and Mann-Whitney U tests. The most frequent GI tumor was lymphoma (58.9%), followed by adenocarcinoma (16.2%) and adenoma (15.0%). Statistical examination revealed that Shiba dogs were predisposed to T-cell lymphoma, Miniature Dachshunds to colorectal B-cell lymphoma and colorectal adenoma, Jack Russell Terriers to adenoma, acinar adenocarcinoma, papillary adenocarcinoma and tubulopapillary carcinoma, French Bulldogs to gastric signet-ring cell carcinoma and plasmacytoma, and Shih Tzus to tubulopapillary adenocarcinoma. These breed predispositions to specific tumors may be unique to the Japanese canine population. To the best of our knowledge, this is the first large-scale epidemiological investigation of canine GI tumors in Japan. The epidemiological information from the present study will serve as a useful reference for clinical veterinarians to establish the differential diagnoses of canine GI tumors.

Diaphragmatic hernia, the protrusion of abdominal organs through a congenital defect or traumatic rupture of the diaphragm, has rarely been documented in wildlife. We report a severe traumatic diaphragmatic hernia in a juvenile female raccoon dog. Postmortem examination revealed a herniation at the Bochdalek site in the right dorsolateral part of the diaphragm, with extensive intestinal and hepatic displacement into the right thoracic cavity. Rib osseous callus indicated healed fractures of the 7th and 8th right ribs. These findings provide an understanding of traumatic diaphragmatic hernias and contribute to the pathological assessment of wildlife health.

A 5-month-old imported boar showed posterior paralysis upon arrival in Japan. Both limbs had coldness, algesia, and thigmesthesia. A thrombus was detected from the abdominal aorta to the left external iliac artery. Irregular vegetation was observed on the left atrioventricular valve. The skeletal muscles in the hind limb appeared like boiled meat. The histological diagnoses were endocarditis, glomerulonephritis, and meningoencephalitis with Gram-negative bacilli as well as diffuse intravascular coagulation and diffuse alveolar damage. The bacilli were identified as Actinobacillus suis via immunohistochemistry and 16S rRNA gene sequencing. This is the first case report of acute posterior paralysis due to iliac thromboembolism in swine with A. suis infection. Iliac thromboembolism should be considered if posterior paralysis is observed in swine.

Postoperative ileus (POI), commonly induced by intestinal manipulation (IM) during laparotomy, results in intestinal wall injury, inflammation, adhesion, and impaired gastrointestinal motility. This study evaluated the effects of mosapride citrate (Mos), a 5-HT₄ receptor agonist, and trehalose (Tre), a cytoprotective disaccharide, on gastrointestinal transit in an IM-induced POI mouse model. Mature male C57BL/6J mice were subjected to IM and allocated into five groups: IM + vehicle (control), IM + Mos (0.2 mg/kg, two times), IM + Tre (100 mg/kg, two times), IM + combined Mos & Tre, and IM + sequential Tre + Mos treatment. Gastrointestinal transit was quantified using phenol red method. All four treatment groups exhibited a significant increase in phenol red recovery in the lower small intestine compared with the control group (P<0.05). Notably, sequential administration (Tre + Mos) resulted in greater dye progression beyond the manipulated region than that of simultaneous (Mos & Tre) treatment (P<0.05). These findings suggest that although both agents individually promote gastrointestinal transit, the therapeutic effect is enhanced when Tre is administered first, followed by Mos. Thus, the timing and sequence of administration play a critical role in the treatment of POI. Sequential Tre + Mos therapy may represent a promising strategy for accelerating the functional restoration of gastrointestinal transit after intestinal injury.

In previous experiments using bullfrog hearts, we successfully replicated both the elevation and reciprocal depression of the ST segment observed in electrocardiograms of human ischemic heart disease. In this study, we induced subepicardial burn injuries in the lateral part of the frog heart ventricle, which allowed us to mimic typical electrocardiographic changes associated with human lateral wall myocardial infarction. These changes included a marked ST segment elevation in the high lateral limb leads (I, augmented Voltage Left; aVL) and reciprocal ST segment depression in the opposite limb leads (III, augmented Voltage Foot; aVF). We surmise that the electrical disparity between injured and intact cardiomyocytes generates currents of injury, leading to the observed ST segment alterations.

Two cats were evaluated because of clinical signs consistent with tetraplegia or ataxia of four limbs and cerebellar signs. The cats were diagnosed with craniocervical junction abnormalities (CJAs) by radiographs, computed tomography and magnetic resonance imaging. Both cats underwent surgical stabilization using a patient-specific titanium atlantoaxial (AA) fixation plate and drill guide templates. In case 2, in addition to AA fixation, the separated dens and the occipital bone compressing the cerebellum were removed. Repositioning and fixation of the AA joint using a custom drill guide template and titanium plate system improved the gait of both cats, but the cerebellar signs persisted. Although CJAs are uncommon in cats, the present cases indicate that surgical stabilization with a custom fixation system may represent a feasible treatment option.

Bovine respiratory disease syndrome, primarily caused by bovine parainfluenza virus type-3 (BPIV-3), is characterized by a high incidence in calves. However, despite its significant clinical importance, the codon usage patterns and evolutionary dynamics of BPIV-3 hosts remain elusive. Hence, this study aimed to systematically analyze the codon usage bias of the BPIV-3 structural protein gene HN, and the roles of mutational pressure and natural selection in its evolution were evaluated. Herein, analysis of indicators such as effective codon number (ENC), relative synonymous codon usage, and codon adaptation index revealed a low codon bias for BPIV-3, with codon preferences showing significant differences from the host. Notably, ENC-GC content at the third codon spot analysis indicated that natural selection dominated codon usage. Additionally, phylogenetic analysis divided BPIV-3 into three main genotypes (namely a, b, and c), with genotype C exhibiting a higher codon adaptability to the host. Altogether, these findings reveal the host–BPIV-3 evolutionary interaction mechanisms, providing a theoretical basis for vaccine design and epidemiological surveillance.

Bovine coronaviruses (BCoVs) cause gastrointestinal and respiratory symptoms in cattle, resulting in economic losses in the industry. The virus strains are endemic in many countries worldwide, including Japan. As recent epidemiological data on BCoVs in Japan are limited, we conducted a survey in Chiba Prefecture, where no such survey has been conducted before, using nasal swab or lung samples collected from 105 cattle with respiratory symptoms during 2020–2022. The rate of BCoV RNA positivity determined using real-time RT-PCR was 43.8% (46 samples), among which 20 strains were successfully isolated. Phylogenetic analysis based on the polymorphic sequence region of the spike gene classified the detected strains into the Japan G3 or G4 genotypes. Therefore, in contrast to previous report that BCoVs of the G3 genotype may have gone extinct in Japan, our results suggest that the Japan G3 and G4 genotypes coexist and have spread in Chiba Prefecture, causing respiratory disease in cattle.

Dengue virus (DENV) is a vector-borne positive-sense RNA virus causing the dengue epidemic in tropical and subtropical regions. Non-structural viral protein 1 (NS1) is a key modulator of host–pathogen interactions. We investigated the in vivo effect of NS1 on DENV infection using an NS1-expressing recombinant vaccinia virus DIs strain (rDIs-DENV2C-NS1). NS1 expression and anti-NS1 antibody induction were observed. To assess its functional role, rDIs-DENV2C-NS1 was administered to AG129 mice lacking type I and II interferon receptors. Two weeks post-immunization, the mice were challenged with DENV2C (10⁵ PFU/mouse). Four days post-infection, the mice immunized with rDIs-DENV2C-NS1 exhibited higher serum viral loads than controls. Liver viral load was significantly increased 14 days after infection compared to that in the control group, and NS1 protein expression was confirmed in the sera. To delineate the effects of NS1, the mice were treated with recombinant NS1 before DENV2 infection. This treatment showed an increase in serum, liver, and spleen viral loads, indicating enhanced viral replication. Serum interleukin (IL)-3, IL-13, IL-18, and macrophage colony-stimulating factor levels were significantly upregulated in the NS1 protein-treated group in a dose-dependent manner. These findings provide in vivo evidences for the immunomodulatory and replication-promoting effects of NS1 during DENV infection.

A 6-year and 11-months-old male striped skunk, Mephitis mephitis, showed inappetence and died. Necropsy revealed multiple hepatic masses, an enlarged spleen, wedge-shaped lesions in the kidneys, and a cervical cyst with white nodules. Histopathologically, the hepatic masses were diagnosed as biliary cystadenomas accompanied by chronic inflammation, the cervical cyst was a thymoma, and the splenic and renal lesions were diagnosed as infarctions. Amorphous eosinophilic material was observed in the liver, spleen, and kidney. These proteinaceous deposits were positive for Congo red staining with yellowish to green birefringence under polarized light. These deposits were immunohistochemically positive for amyloid A (AA). This report describes the pathological features of AA amyloidosis, biliary cystadenoma and cervical thymoma in a striped skunk.

Chitin digestion in pangolins and other anteaters is thought to be aided by commensal bacteria in the digestive tract, in addition to their chitinase. This study characterized the gut microbiota of captive Javan pangolins using amplicon sequencing. Fecal samples were collected from two individuals and were sampled twice over one week. The dominant bacterial phyla identified were Firmicutes (Bacillota), Bacteroidetes (Bacteroidota), Proteobacteria (Pseudomonadota), and Actinobacteria (Actinomycetota). The most prevalent genera included Clostridium, Bacteroides, Lactobacillus, Bifidobacterium, Streptococcus, and Sporosarcina. Alpha and beta diversity were relatively low between paired samples, but the short sampling interval limits conclusions about microbial stability. These findings provide insights into the Javan pangolin’s gut microbiota and support future research on microbial contributions to their digestion, health, and conservation.

Urinary calculus is common but often unnoticed condition in African spurred tortoises (Centrochelys sulcata) until severe, challenging early detection and etiology. This case uniquely features detailed, prolonged home care by the owner-veterinarian. A 13-year-old male presented with obstipation due to a 10-cm urolith. Home management stabilized him for surgical removal via cystotomy. Postoperatively, the patient developed chronic hyperuricemia and gout, leading to progressive emaciation and death two years later. Autopsy revealed systemic articular and visceral gout, severe chronic cystitis, and renal fibrosis. This case underscores calculi grow massive before detection, often with patient deterioration. This detailed report, including comprehensive autopsy findings, provides novel longitudinal data the crucial role of appropriate supportive care in stabilizing the patient for surgical intervention following the detection of massive urolithiasis, while also linking between successful surgical removal and the subsequent development of systemic gout, emphasizing the critical need for long-term therapeutic monitoring of uric acid metabolism in chelonians post-urolith removal for all stakeholders involved in the care of this species.