We examined the relationship between inherited motor-related conformation and orientation of facial hair whorls in Japanese Kiso horses. Eleven horses were divided into clockwise, counterclockwise, and radial groups according to facial hair whorls. We placed six markers on anatomical landmarks of each lateral side in a horse and measured the height of the landmarks, the distance between adjacent landmarks, and the angle of the adjacent landmarks. In the counterclockwise group, the horses tended to exhibit higher values on the left side than on the right side, and the comparison of the height of landmarks revealed a significant difference between both sides. Therefore, the orientation of facial hair whorls may suggest the tendency of motor-related conformation, at least in counterclockwise group.

Clostridium sp. was detected in the organs of a cow with black watery diarrhea in Japan. Results identifying this species were inconsistent; Clostridium novyi type A infection was suggested by PCR assay targeting Clostridium fliC region (fliC-multiplex PCR), while 16S rRNA gene sequencing identified the isolated bacteria as Clostridium massiliodielmoense. Sequencing of fliC-multiplex PCR products from the isolates revealed the presence of fliC region in C. massiliodielmoense, which had 92.7% nucleotide similarity to that of C. novyi type A JCM 1406^T, leading to the false positive detection of C. novyi by the PCR. This is the first C. massiliodielmoense isolation from clinical specimens, suggesting the need for further research on its pathogenicity and improvement in fliC-multiplex PCR.

In equine regenerative medicine using bone marrow-derived mesenchymal stem/stromal cells (BM-MSC), the importance of the quality management of BM-MSC has been widely recognized. However, there is little information concerning the relationship between cellular senescence and the stemness in equine BM-MSC. In this study, we showed that stemness markers (NANOG, OCT4, SOX2 and telomerase reverse transcriptase) and colony forming unit-fibroblast apparently decreased accompanied with incidence of senescence-associated β-galactosidase-positive cells by repeated passage. Additionally, we suggested that down-regulation of cell proliferation in senescent BM-MSC was related to increased expression of cyclin-dependent kinase inhibitor 2B (CDKN2B). On the other hand, forced expression of NANOG into senescent BM-MSC brought upregulation of several stemness markers and downregulation of CKDN2B accompanied with restoration of proliferation potential and osteogenic ability. These results suggested that expression of NANOG was important for the maintenance of the stemness in equine BM-MSC.

Noma horses are native Japanese horses. Health checkups revealed that many Noma horses developed dermatitis during summer, which subsided in winter. Seasonal development and signs of itching, suggestive of allergic dermatitis, were observed. In this study, allergen-specific IgE was measured using blood samples collected from 15 Noma horses in summer and winter to identify allergens highly associated with dermatitis. The presence of dermatitis in the subject individuals was recorded during blood sample collection. White blood cell and eosinophil counts, serum total IgE concentration, and serum allergen-specific IgE units (ARUs) were measured. White blood cell and eosinophil counts were significantly higher in horses with dermatitis in summer compared to winter. On the other hand, there was no significant difference in serum total IgE concentration regardless of the presence of dermatitis or the season. Horses with dermatitis in summer showed higher ARUs derived from red ants, horseflies, biting midges, cockroaches, deerflies, and mosquitoes than those in winter. These ARUs were positively correlated with white blood cell and eosinophil counts. The factor analysis results suggested that sensitization to some insects, such as mosquitoes and deerflies, may be a cause of dermatitis. In conclusion, insect-derived allergens could be associated with the onset of dermatitis in Noma horses.

Three Shiba goats aged 1 to 7 years kept in Ibaraki prefecture in Japan were presented with chief complaint of lumbar paralysis or gait abnormalities. As cerebrospinal setariasis were suspected in all cases at the first stage, ivermectin was administered to treat, but the response was insufficient. Necropsy revealed abscess formation on the ventral side of the spine at T5 in Case 1, T5–6 in Case 2, and C7–T1 in Case 3, causing compression of the spinal cord in all three cases. In addition to cerebrospinal setariasis, vertebral abscess should be considered as a cause of paresis or gait abnormalities in goats in Japan. Computed tomography was a useful for diagnosing vertebral abscess.

Forty-two Japanese Black cattle on two farms in Kagoshima prefecture, Japan were used in this study. The rearing style of farm A was in a dark barn with a large roof to block sunlight (n=21). The rearing style of farm B was grazing, and exposed to direct sunlight (n=21). Blood sampling was performed twice on the same cattle in August 2022 (summer) and February 2023 (winter). As the results, the serum 25(OH)D₃ concentrations were significantly lower in cattle of farm A than in cattle of farm B (P<0.01), and were significantly lower in winter season than in summer season (P<0.01). These results showed that there were differences in blood 25(OH)D₃ concentrations between the farms or seasons.

A 5-year-old Japanese Black cow presented with astasia. Bovine leukemia virus (BLV) was detected in the peripheral blood with lower proviral load (PVL). No enlargement of surface lymph nodes or lymphocytosis was observed. Necropsy revealed no enlarged lymph nodes in the thoracic, abdominal, or pelvic cavity. Spinal epidural and peri-medullary adipose tissue was increased in the spinal canal of lumbar to sacral vertebrae, Histopathological examination revealed tumor invasion of the epidural adipose tissue, and a diagnosis of B-cell lymphoma was made. The PVL in tumor tissue was higher, and monoclonal integration of BLV was confirmed. It was a rare case of bovine enzootic leukosis that formed a solitary mass around the spinal cord which might cause hindlimb paresis.

The inactivated vaccine is effective in controlling foot-and-mouth disease (FMD), but it has drawbacks such as the need for a biosafety level 3 laboratory facility to handle live foot-and-mouth disease virus (FMDV), high production costs, and biological safety risks. In response to these challenges, we developed a new recombinant protein vaccine (2BT-pIgG-Fc) containing porcine IgG-Fc to enhance protein stability in the body. This vaccine incorporates two-repeat B-cell and one-single T-cell epitope derived from O/Jincheon/SKR/2014. Our study confirmed that 2BT-pIgG-Fc and a commercial FMDV vaccine induced FMDV-specific antibodies in guinea pigs at 28 days post-vaccination. The percentage inhibition (PI) value of 2BT-pIgG-Fc was 90.43%, and the commercial FMDV vaccine was 81.75%. The PI value of 2BT-pIgG-Fc was 8.68% higher than that of commercial FMDV vaccine. In pigs, the primary target animals for FMDV, all five individuals produced FMDV-specific antibodies 42 days after vaccination with 2BT-pIgG-Fc. Furthermore, serum from 2BT-pIgG-Fc-vaccinated pigs exhibited neutralizing ability against FMDV infection. Intriguingly, the 2BT-pIgG-Fc recombinant demonstrated FMDV-specific antibody production rates and neutralization efficiency similar to commercial inactivated vaccines. This study illustrates the potential to enhance vaccine efficacy by strategically combining well-known antigenic domains in the development of recombinant protein-based vaccines.

The prevalence of chicken coccidiosis in the poultry industry is a significant concern, further exacerbated by the emergence of drug-resistant coccidia resulting from the indiscriminate use of medications. Ethanamizuril, a novel triazine anti-coccidial compound, has been used to combat drug resistance. Currently, it is known that Ethanamizuril acts on the second-generation merozoites and early gametogenesis stages of Eimeria. Limited information exists regarding its impact on the early merozoites and exogenous stage of Eimeria. In the present study, the anti-coccidial properties of Ethanamizuril were evaluated both in vitro and in vivo. The in vitro experiments demonstrated that Ethanamizuril effectively inhibits the sporulation of E. tenella oocysts in a dose-dependent manner and significantly reduces the sporozoite excystation rate. Furthermore, in vivo tests revealed that treatment with 10 mg/L Ethanamizuril in drinking water significantly decreased the copy number of first-generation and secondary-generation merozoites in the chicken cecum, indicating that it can inhibit the development of whole schizonts development. Moreover, treatment with Ethanamizuril demonstrated excellent protective efficacy with an anti-coccidial index (ACI) of 180.2, which was manifested through higher body weight gains, lighter cecal lesion, lower fecal oocyst shedding score and reduced liver index. Collectively, this study suggests that Ethanamizuril effectively treats E. tenella infection by inhibiting both endogenous and exogenous stages development.

The bear roundworm Baylisascaris transfuga has been identified in several host bears (Ursinae). However, limited genetic information is available on the bear roundworm in Japanese populations. This study evaluated the genetic composition of bear roundworms isolated from wild Japanese black bears indigenous to Lake Towada, Japan. First, we conducted genetic and/or molecular phylogenetic analyses based on cytochrome c oxidase subunit II and internal transcribed spacer 2 among Baylisascaris species. These analyses revealed that the identified roundworms were genetically B. transfuga. In addition, the average body size of the obtained roundworms in this study was almost the same as that previously reported for B. transfuga. This study represents an important step in genetic research on the roundworm B. transfuga in Ursinae bears not only from Japan but also from the rest of the world.

Malakoplakia is a rare granulomatous inflammation that has mainly been reported in the urinary bladder of dogs. Only one case of canine colonic malakoplakia has been reported to date; however, successful treatment of this disease has not been reported. Here, we report a case of colonic malakoplakia in a 5-month-old spayed female French Bulldog. The dog was referred to a veterinarian because of chronic diarrhea and mucinous blood feces; empirical treatment did not improve its condition. Histologically, numerous macrophages containing periodic acid-Schiff-positive granules infiltrated the lamina propria of the large intestine. Furthermore, targetoid basophilic inclusion bodies (Michaelis-Gutmann bodies) were observed. Complete clinical remission was achieved after 8 months of enrofloxacin treatment and favorable progress after 2 months of medication.

Chlorogenic acid (CGA) is a polyphenol substance contained in many plants, which has good antioxidant activity. This experiment aimed to explore the protective effects of CGA on hydrogen peroxide (H₂O₂)-induced inflammatory response, apoptosis, and antioxidant capacity of bovine intestinal epithelial cells (BIECs-21) under oxidative stress and its mechanism. The results showed that compared with cells treated with H₂O₂ alone, CGA pretreatment could improve the viability of BIECs-21. Importantly, Chlorogenic acid pretreatment significantly reduced the formation of malondialdehyde (MDA), lowered reactive oxygen species (ROS) levels, and enhanced the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) (P<0.05). In addition, CGA can also improve the intestinal barrier by increasing the abundance of tight junction proteins claudin-1 and occludin. Meanwhile, CGA can reduce the gene expression levels of pro-inflammatory factors Interleukin-6 (IL-6) and Interleukin-8 (IL-8), increase the expression of anti-inflammatory factor Interleukin-10 (IL-10), promote the expression of the nuclear factor-related factor 2 (Nrf2) signaling pathway, enhance cell antioxidant capacity, and inhibit Nuclear Factor Kappa B (NF-κB) the activation of the signaling pathway reducing the inflammatory response, thereby alleviating inflammation and oxidative stress damage.

Microsporidia can cause infection in various animals and humans. To determine the recent prevalence of Encephalitozoon in companion birds in Japan, 364 bird feces and 16 conjunctival exudates, as well as 28 exhibition bird feces, were examined using polymerase chain reaction (PCR). Thirty-five (9.6%) feces and 2 (12.5%) conjunctival exudates from companion birds were PCR positive, and sequence analysis revealed that all detected organisms were Encephalitozoon hellem genotype 1A. The prevalence by region varied from 4.5% in the Shikoku region to 14.3% in the Chugoku region. By age, the prevalence in birds younger than 6 months of age was 13.3%. We also discuss the threat of human infection as a zoonotic disease.

No study has determined the minimal effective dose of trazodone required to induce behavioral changes and its safety profile in rabbits. Therefore, this study aimed to determine the minimal effective dose of trazodone to improve compliance to handling, and to evaluate associated changes in motor activity, physiological and arterial blood gas parameters. Eight intact female New Zealand White rabbits (2-month-old; 1.66 ± 0.12 kg) were included in this prospective, blinded, randomized cross-over study. After a 10-day acclimation, rabbits randomly received placebo or trazodone 10, 20 or 30 mg/kg orally (PLAC, TRAZ10, TRAZ20, TRAZ30) with a 1-week wash-out period. Compliance scoring (dynamic interactive visual analog scale; DIVAS), activity levels measured with accelerometry (T0-T600), physiological parameters (temperature, heart, and respiratory rates), and arterial blood gas parameters (up to T240) were evaluated. Compliance scores, accelerometry, physiological and arterial blood gas parameters and hypoxemia prevalence (PaO₂ <60 mmHg) were analyzed using linear mixed models and Chi-squared tests, respectively (P<0.05). When compared with PLAC, DIVAS scores were significantly higher at T80-120, T40-120 and T120-200 in TRAZ10, TRAZ20 and TRAZ30 post-administration, respectively. When compared with baseline, DIVAS scores were significantly higher from T80-160, T40-240 and T80-200 in TRAZ10, TRAZ20 and TRAZ30, respectively. All other parameters were not significantly different. In TRAZ30, hypoxemia was observed in 2/8 rabbits (P=0.104). In conclusion, oral trazodone improved rabbit compliance at all studied dosages, especially 20 mg/kg improved rabbit compliance without decreasing motor activity or causing hypoxemia.

Canine urothelial carcinoma (cUC) is one of the most malignant tumors affecting dogs; however, its proliferative mechanism is yet to be fully elucidated. The ubiquitin-proteasome system (UPS) is an important metabolic pathway regulating protein degradation, and its dysfunction leads to apoptosis. We investigated the antitumor effect of the proteasome inhibitor bortezomib, which blocks the UPS. Bortezomib inhibited cell growth in cUC cell lines by inducing apoptosis in vitro. These findings suggest the potential of bortezomib as a novel therapeutic drug for dogs with cUC.

Fesaviruses, picorna-like RNA viruses, were discovered in 2014 in feces from cats in an animal shelter in the United States but have not since been reported elsewhere. In this study, we collected cat fecal samples from 20 adult cats from an animal shelter in Tokyo, Japan, and examined them for viral pathogens. Next generation sequencing (NGS) was performed to detect both RNA and DNA virus sequences. Sequences of a total of 7 RNA viruses including some common feline pathogenic viruses were detected across 8 samples, while no DNA virus sequences were identified in any sample. Of the RNA virus sequences detected in the samples, two sequences, 4,746 and 4,439 bp, demonstrated 90.3% and 85.0% similarity, respectively, to the fesavirus 4 sequence in the database. To confirm the NGS results, quantitative RT-PCR (qRT-PCR) assays were developed using specific primers and probes designed based on the contig sequences. Based on the qRT-PCR assays, we detected relatively high copy-numbers of fesavirus 4 RNA in the two fecal samples from which the fesavirus 4 sequences were originally obtained, and low copy numbers in other samples. These results demonstrate the presence of fesavirus 4 in cats in Japan for the first time.

Feline parvovirus (FPV) or feline panleukopenia virus is a highly contagious, life-threatening infectious virus in cats. Although FPV vaccination is routinely practiced in China, clinical diseases continue to occur. The investigation of genotypes and viral evolution can contribute to the prevention, diagnosis, and treatment of FPV. Therefore, this study aimed to provide an up-to-date understanding of the epidemiological, genotypic, and phylogenetic characteristics of FPV. In total, 152 rectal swabs were collected from diseased cats. All swab samples were tested for FPV using molecular methods. Amplification of the complete viral protein 2 (VP2) gene was performed for further analysis and to infer the genotypic and evolutionary characteristics of FPV. Of the 152 samples, FPV DNA was detected in 17 (17/152, 11.18%). Cats with FPV showed variable clinical signs such as dehydration, anorexia, fever, vomiting, and blood-stained diarrhea. Furthermore, VP2 sequences were identified in 17 PCR-positive cats, confirming the presence of FPV. Phylogenetic and nucleotide pairwise identity analyses revealed high genetic similarity among FPV sequences (99.6–100%) and clustered them into the FPV-G1 group. Amino acid analysis indicated a novel mutation (Ala91Ser) in all VP2 gene sequences amplified in this study. Our study provides baseline epidemiological data for the better prevention of FPV with respect to vaccination strategies. Genotypic and phylogenetic analyses confirm that FPV-G1 was the predominant FPV group in infected cats in Kunshan. Therefore, a rigorous countrywide investigation of the genotypic and evolutionary characteristics of FPV is warranted.

Fecal corticosterone metabolite (FCM) concentrations, which can be determined noninvasively, have recently been explored as a stress indicator in birds. In our study, we measured FCM concentrations in penguins under nonmolting or molting conditions, cool or hot season, diseased condition, and incubation period. These measurements were conducted in an aquarium that housed king penguins, gentoo penguins, and African penguins. This study aimed to investigate the validity of fecal matter as a stress indicator. Our findings revealed that FCM concentrations were significantly higher in molting individuals than in nonmolting individuals. Compared with the cool season, FCM concentrations were significantly higher in penguins housed outdoors during the hot season. However, no differences were observed in penguins housed indoors. Diseased individuals and an incubating individual showed notably higher FCM concentrations than healthy individuals. Interestingly, the FCM concentration in king penguin that underwent cataract surgery was extremely high before the surgery. However, 1 month postsurgery, it decreased to a level similar to that of healthy individuals. We observed increased FCM concentrations in penguins considered to be exposed to stressors. Notably, FCM concentration decreased after removing the stress factor. The FCM concentration was found to be consistent with the stress state of penguins, suggesting its usefulness as a stress indicator.

There are currently no standard methods for diagnosing cardiac diseases in dolphins. These diseases may consequently be overlooked and go undiagnosed. The presence and severity of cardiac diseases in humans can be determined using blood tests. Atrial natriuretic peptide (ANP) used in human cardiac examinations has low species specificity. There have already been reports of homology between dolphin and human ANP; however, its potential for clinical application in dolphins has not been tested. This study was conducted to establish a reference for ANP levels in healthy bottlenose dolphins. Healthy bottlenose dolphins (seven females; estimated to be 7–30 years of age) at an aquarium in Japan were sampled. Each animal was tested for ANP at least three times, and the mean value and standard deviation were calculated to be 43.4 ± 19.2 pg/mL. In humans, patients with high plasma ANP levels have a poor prognosis. In veterinary medicine, cutoff values for the diagnosis of mitral regurgitation and heart failure in dogs have been established and used to predict prognosis. The results of the present study may contribute to the health management of bottlenose dolphins, particularly in the early detection and treatment of cardiac diseases.