Journal of Veterinary Medical Science最新文献

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.

Neutrophil recruitment to inflamed tissue is mediated by adhesion molecules and chemokine signaling. Fuzapladib sodium hydrate (FZP) has been reported to reduce neutrophil infiltration, but its broader effect on adhesion molecules in dogs remains unclear. This study investigated the influence of FZP on canine aortic endothelial cells (CnAOEC) and primary neutrophils under cytokine stimulation. CnAOEC were treated with recombinant canine tumor necrosis factor alpha (rh-cTNFα) or lipopolysaccharide together with FZP, while neutrophils isolated from healthy dogs were preincubated with FZP before rh-cTNFα stimulation for gene and protein expression analyses. Expression of endothelial E-selectin, P-selectin, and intercellular adhesion molecule-1 (ICAM-1) was quantified by qPCR. Neutrophil C-X-C motif chemokine receptor 2, P-selectin glycoprotein ligand-1 (PSGL-1), and L-selectin were also analyzed using qPCR. In addition, C-X-C motif chemokine ligand 1 and leukocyte function-associated antigen-1 (LFA-1) were determined using ELISA and flow cytometry, respectively. Neutrophil adhesion to FZP-treated endothelial monolayers was evaluated under static conditions. FZP significantly downregulated endothelial P-selectin at the highest dose, whereas ICAM-1 was upregulated. In neutrophils, FZP decreased L-selectin and PSGL-1 expression at the highest concentration. These findings suggest that FZP selectively modulates early adhesion through suppression of P-selectin, L-selectin, and PSGL-1, which may weaken rolling and tethering interactions between cells. Although functional adhesion effects were limited, these molecular responses provide further insight into the anti-inflammatory effect of FZP.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL), a common neoplastic disease in cattle. Since animals with high BLV proviral load (PVL) pose a greater transmission risk, identifying cattle with low PVL is crucial for herd management. Certain bovine leukocyte antigen (BoLA)-DRB3 alleles- DRB3*009:02, DRB3*014:01:01, and DRB3*002:01-are strongly associated with low PVL in Holstein cattle and thus can serve as markers for genetic resistance. In this study, we developed a rapid, cost-effective allele-specific PCR (AS-PCR) assay using sequence-specific primers to detect these three alleles. The assay was validated through both synthesized DNA and genomic DNA samples from pre-genotyped cattle, showing 100% sensitivity and 94% specificity compared with PCR-sequence-based typing (SBT). The detection limit was 0.8 ng/reaction. A two-stage pooling approach allowed high-throughput screening of field samples, and analysis of 444 cattle revealed that 27.0% carried at least one target allele. Considering only standard PCR and agarose gel electrophoresis are required, this assay is accessible to basic laboratories without advanced equipment or bioinformatics expertise. Moreover, it enables rapid genotyping of BLV resistance markers, offering a practical tool for genetic selection and disease control in dairy farms, especially those in high-BLV-prevalence regions.

Trapped Neutrophil Syndrome (TNS) is a rare, fatal, autosomal recessive disease in Border Collies. A 7-month-old female Border Collie with TNS presented with fever, diarrhea, occasional vomiting, lameness, and severe neutropenia. The diagnosis was confirmed by bone marrow examination and vacuolar protein sorting-associated protein 13B (VPS13B) gene mutation analysis. Initial treatment with prednisolone improved symptoms; however, relapse occurred during tapering, and the addition of cyclosporine resolved the symptoms, allowing successful tapering of prednisolone. The dog remained symptom-free for 7.4 months and survived for 33 months. Cyclosporine may be associated with increased C-X-C chemokine receptor type 4 mRNA expression, possibly reducing the risk of infection and fever. Based on these findings, a combination of cyclosporine and prednisolone may represent a potential treatment option for TNS.

We previously reported that poly (N-vinylacetamide-co-acrylic acid) coupled with d-octaarginine (VP-R8) efficiently introduces proteins and nucleic acids into cells. Based on these results, we hypothesized that VP-R8 can introduce a complex composed of guide RNA and Cas9 (RNP complex) into cells and induce genome editing mediated by the CRISPR-Cas9 system. We tested this hypothesis using a mouse dendritic cell line and mouse primary dendritic cells. The RNP complexes formed by guide RNA consisting of CRISPR RNA (crRNA), fluorescently labeled trans-activating crRNA (tracrRNA), and GFP-fused Cas9 were introduced into a mouse dendritic cell line using VP-R8 or control transfection reagents. Cells transfected using VP-R8 exhibited higher fluorescence than those transfected with other transfection reagents, indicating that VP-R8 efficiently introduced the RNP complex into the mouse dendritic cell line. Genome editing of the target DNA was detected in cells transfected with the RNP complex using VP-R8 and dominant relative to other transfection reagents. We also observed that VP-R8 effectively delivered RNP complexes consisting of single-guide RNA and Cas9 and induced genome editing in the dendritic cell line. Additionally, VP-R8 efficiently delivered RNP complexes into mouse primary dendritic cells and induced genome editing of the functional gene without producing early inflammatory cytokines. Thus, VP-R8 shows potential as a transfection tool to generate dendritic cells with specific gene regions deleted by genome editing via the CRISPR-Cas9 system. This approach aims to elucidate the detailed molecular mechanisms of dendritic cell function for its application to vaccines.

Male Sulawesi babirusas (Babyrousa celebensis) are known for their distinctive backward-curving canine tusks. Previous research on the anomalies of canines in male babirusa skulls has demonstrated a high prevalence of growth abnormalities due to failure in the rotation of the alveolar process and the curvature of the canines, while the numerical abnormalities were found to be rare. This study focused on supernumerary maxillary canines as observed in a wild-caught babirusa in an ex situ conservation program. This report represents the first publication of photographic evidence of a living babirusa with three right maxillary canines, one left maxillary canine growing toward the right side of the snout, and two overgrown mandibular canines threatening to erode the forehead.

The present study investigated whether behavioral changes predict the onset of left displaced abomasum (LDA) using a small, multi-sensor device attached to a cow collar that records data in real time over a 24 hr period. Behavioral data were collected from 17 LDA cows that had undergone surgery and 23 healthy dairy cows. The total rumination time over 10 days before and after LDA surgery in cows, was significantly shorter from 8 days before to 3 days after surgery, peaking at 216.4 ± 96.8 min on day before surgery than that in the control group (group: P<0.001). The total rumination time of LDA cows was significantly shorter than that of the control group at each time point (interaction effect: P<0.001). The diagnostic ability to predict the onset of LDA based on the cutoff value (≤360 min) of total rumination time 6 days before LDA surgery calculated using the Yohden index showed a significantly high AUC of 0.887 (P<0.0001) with high sensitivity (84.6%) and specificity (88.9%). In other words, it was suggested that the onset of LDA can be predicted by total rumination time from 6 days before surgery. An Animal Health Monitoring System (AHMS) not only warns of the onset of cattle diseases, such as LDA, several days in advance, but may also predict the onset of diseases up to 6 days in advance based on shortened the total rumination times, thereby contributing to the efficiency of work for farmers and veterinarians.

In this study, we analyzed the prevalence of diabetes in dogs over a nine-year period (2015-2023) in Japan using records provided by a pet insurance company. The observed prevalence rose from approximately 0.07% in 2015 to 0.30% in 2023, highlighting a steady upward trend. Logistic regression analysis confirmed this increase (regression coefficient β=0.121, P<0.001), with an odds ratio of 1.128 (95% confidence interval: 1.116-1.142), indicating an annual 12.8% increase in the odds of diabetes. A similar trend was observed among newly enrolled animals each year (β=0.0927, P<0.001; OR=1.097, 95% confidence interval: 1.076-1.119), suggesting a 9.7% annual increase in the odds of developing diabetes. The insurance dataset consistently showed a higher prevalence in males compared to females across all years. Additionally, we examined 209 clinical cases of canine diabetes collected from 21 veterinary hospitals to investigate age at diagnosis, sex, and neutering status. Case records indicated an age-related increase in diabetes incidence, with a peak between 8 and 10 years of age. The timing of diabetes onset did not significantly differ by sex or neutering status (χ² test, P=0.936). These findings highlight a progressive increase in canine diabetes prevalence in Japan and support the existence of a sex-based disparity in risk, with males being more susceptible.

Equine rhinitis B virus (ERBV), a member of the Picornaviridae family, is associated with mild to moderate respiratory illness in horses, yet its genomic diversity remains incompletely characterized. In this study, we performed metagenomic analysis on a rectal swab sample from a diarrheic foal confirmed to be positive for rotavirus A. Unexpectedly, we identified a highly divergent ERBV strain with a 9,448-nucleotide genome encoding a 2,721-amino-acid polyprotein. Sequence analysis revealed only 62.5-63.1% identity in the polyprotein and 47.1-49.8% in the VP1 region compared to known ERBV serotypes, suggesting a novel genotype. The genome exhibited typical picornavirus features, including a type II internal ribosome entry site (IRES), but also unique elements such as an 87-amino-acid insertion in the leader proteinase region and atypical cleavage motifs. A strain-specific RT-qPCR assay was developed alongside a broadly reactive assay targeting ERBV1-3. Screening of 37 rectal swab samples from horses revealed a 10.8% positivity rate using the new assay, with clustered cases from the same farm in 2022 and an additional case in 2024. No samples were positive using the broad-range primers, indicating potential gaps in surveillance. These findings highlight the importance of metagenomics in pathogen discovery and underscore the need for updated molecular tools to monitor genetically distinct ERBV strains and assess their clinical significance.

The large Japanese field mouse, Apodemus speciosus, is a rodent species endemic to Japan and is widely distributed throughout the country. It has been extensively used as a model organism in studies of geographic isolation and as an environmental bioindicator. However, there have been no reports on the establishment of pluripotent stem cells from this species that could be applied to developmental engineering. In this study, we attempted to generate induced pluripotent stem cells (iPSCs) from A. speciosus and successfully obtained cells that formed dome-shaped colonies and expressed pluripotency markers. These cells were capable of forming embryoid bodies that expressed markers of the three germ layers, and in vivo analysis using immunodeficient mice confirmed their ability to form teratomas. A. speciosus iPSCs established in this study not only contribute to the conservation of Japan's valuable animal genetic resources, but also provide a novel platform for investigating various biological processes that remain unexplored in this wild species.