Little is known about the vomeronasal organ (VNO), which is associated with social behaviors through pheromone detection in mammals, particularly ungulates. We investigated the distribution of phospholipase C beta 2 (PLCβ2), a marker of solitary chemosensory cells (SCCs), in the VNO of the Korean roe deer (Capreolus pygargus). PLCβ2-positive cells were detected in both the sensory and non-sensory epithelium of the VNO, and resembled the bipolar cells. PLCβ2-positive cells colocalized with choline acetyltransferase, and Lens culinaris agglutinin. Furthermore, PLCβ2-positive cells were surrounded with trigeminal nerve fibers with calcitonin gene-related peptide (CGRP)-positive immunoreaction. The results indicate that the VNO of the Korean roe deer exhibits a distinct distribution of SCCs, which may be involved in detecting risk factors such as harmful substances.

In 2024, 90 soil samples and 11 fecal samples were collected from nine Mongolian provinces. Using NANAT selective agar, R. equi was successfully isolated from 23 soil samples (25.6%) across five provinces and from three fecal samples (27.3%) collected in two provinces. A total of 122 isolates were identified as R. equi via choE-targeted polymerase chain reaction (PCR) and subsequently screened for virulence-associated genes (vapA, vapB, and vapN) by PCR. Of these, 17 isolates tested positive for the vapA gene, while the remaining 105 isolates were negative for both vapB and vapN. Plasmid profiling of the vapA-positive isolates revealed the presence of an 85-kb type I virulence plasmid, which is common in isolates from Europe and North America. This is the first documented detection of vapA-positive R. equi in Mongolia.

A 14-year-old, castrated male mixed-breed cat presented with weight loss and rhinitis for several months and was referred due to anemia and thrombocytopenia. A complete blood cell count (CBC) revealed severe non-regenerative anemia (hematocrit (HCT), 9.7%) and thrombocytopenia (0 /μL). Serology was positive for feline immunodeficiency virus (FIV) antibodies. Bone marrow aspiration showed moderate myeloid hyperplasia, severe erythroid hypoplasia, and megakaryocytic hypoplasia. The cat was diagnosed with pure red cell aplasia (PRCA) and amegakaryocytic thrombocytopenia (AMT) and treated with corticosteroids, leading to recovery. On day 242, bone marrow aspirates showed erythroid and megakaryocytic cell production. By day 600, both diseases had not recurred without medication. This is the documented case of concurrent PRCA and AMT in an FIV-infected cat.

Epizootic hemorrhagic disease virus (EHDV) is a vector-borne pathogen that affects both wild and domestic ruminants. Climate influences vector-borne diseases by driving vector migration to new areas, where they spread the virus. However, the lack of surveillance in some areas hinders accurate assessment the true disease burden. This study aimed to determine the seroprevalence of EHDV and associated risk factors in cattle from the northern region of Peru in 2022. Blood serum samples were collected from 578 cattle in the departments of Tumbes, Piura, Lambayeque, Cajamarca, and La Libertad and analyzed using cELISA. The overall EHDV seroprevalence was 17.82% (95% confidence interval 14.78–21.19). The highest proportion of seropositive animals was observed in two deparments closest to the equator, Piura and Tumbes (50%), followed by Lambayeque (39.1%) and Cajamarca (11.08%). In addition, the highest seroprevalence was found at temperatures ≤20°C (50.46%), wind speeds ≤3 m/sec (31.43%), and altitudes ≤1,260 masl (37.28%), with a significant decrease at higher elevations (P<0.05). After adjusting for temperature, the odds of EHDV seropositivity were significantly lower in cattle from areas located at >2,000 to ≤3,290 masl (Odds ratio [OR]=0.15) and >3,290 (OR=0.07), compared to those from the reference altitude category of ≤1,260 masl. Similarly, after adjusting for altitude, cattle from deparments with temperatures >20°C had significantly lower odds of seropositivity (OR=0.17) compared to those exposed to temperatures ≤20°C. This study reports anti-EHDV antibodies in cattle from northern Peru for the first time, highlighting associations with bioclimatic factors.

To diagnose milk fever (MF) in dairy cows under clinical practice, measurement of ionized calcium concentrations in blood using point-of-care (POC) devices is required. However, blood sampling poses a significant financial burden on farmers and increases the workload of veterinarians who must visit farms to perform the examinations. The present study aimed to evaluate whether the onset of MF could be predicted using colostrum samples, which can be collected by farmers themselves. Total calcium (tCa) concentrations in colostrum were measured using a commercially available water quality test device. The results showed that cows treated for MF had significantly higher tCa concentrations in colostrum (2,141.1 ± 623.7 mg/L) compared to cows without MF (1,546.3 ± 494.4 mg/L) (P=0.006). Receiver operating characteristic (ROC) curve analysis suggested that measuring tCa concentration in colostrum could be used to predict the onset of MF. When the cut-off value was set at >2,000 mg/L, the sensitivity and specificity were 66.7% and 88.9%, respectively. Results from these findings, measuring tCa concentration in colostrum using a water quality test device offers a simple, inexpensive, and rapid method for predicting the onset of MF in dairy cows.

Peripheral nerve sheath tumors (PNSTs) originating from the vagus nerve are rare in dogs. This paper reports a canine case of malignant PNST originating from the vagus nerve, specifically the vagus-sympathetic trunk, which induced notable gagging and chronic vomiting. The clinical signs improved after surgery despite incomplete resection, and the dog achieved an overall survival of at least 514 days. These observations suggest that surgical intervention may be beneficial in dogs with malignant PNSTs originating from the vagus nerve.

A 13-year-old female Persian cat with a heart murmur presented with dyspnea. Thoracic radiography revealed pleural effusion and dilation of peripheral pulmonary vessels. Echocardiography showed significant enlargement of the right atrium and ventricle and left atrial appendage; an atrial septal defect (ASD) near the atrioventricular valves; and a septum-like structure below the left atrial appendage. Left-to-right shunting through the ASD was noted, with a pulmonary-to-systemic flow ratio of 2.72. The cat was diagnosed with incomplete atrioventricular septal defect (IAVSD) and supravalvular mitral stenosis (SVMS) and subsequently medically managed for 7 months before death. Necropsy confirmed IAVSD with a mitral valve cleft and SVMS. To our knowledge, this is the longest reported survival in a cat with both anomalies.

We aimed to establish a streptozotocin-induced type 1 diabetes mellitus (DM) model in Microminipigs. Initially, a dose of 150 mg/kg streptozotocin was tested, and pigs exhibited characteristic triphasic patterns in glycemic responses: phase 1 (transient hyperglycemia), phase 2 (hypoglycemia), and phase 3 (sustained hyperglycemia). Blood glucose concentrations showed a transient increase in phase 1, and then dropped sharply in phase 2. Based on these findings, we conducted a follow-up study using a reduced streptozotocin dose of 125 mg/kg. This group exhibited glucose dynamics similar to those observed in the 150 mg/kg group, showing glucose reductions and mild lethargy. In phase 3, both groups developed sustained hyperglycemia with negligible insulin secretion, confirming the successful establishment of a type 1 DM model.

Eimeria spp. cause coccidiosis characterized by diarrhea and induce serious economic losses in livestock industries. Although several anti-coccidial drugs are currently available, the emergence of resistant strains and drug residues is problematic; therefore, the development of new drugs is needed. Since sporozoites of Eimeria spp. invade host intestinal epithelial cells and numerous merozoites are formed, drugs that target sporozoites are expected to be useful. We previously used murine Eimeria krijgsmanni as a model to examine anti-coccidial drug susceptibility; however, few studies have conducted drug evaluations against sporozoites. The establishment of excystation protocols is essential for progress in in vitro experiments using sporozoites because oocysts must be isolated from feces using complex techniques before the excystation process. Various artificial excystation protocols have been reported for each Eimeria spp.; however, those for E. krijgsmanni have not yet been examined. Therefore, 4 protocols described in previous studies were herein conducted for E. krijgsmanni. Pepsin was important for excystation in rodent Eimeria spp., and this was also the case for E. krijgsmanni. Excystation rates were higher with the physical disruption of oocyst walls than with pepsin. An incubation in HBSS containing 0.25% (w/v) trypsin and 0.1% (w/v) sodium taurocholate after a physical treatment achieved higher and the most stable excystation rates. Modifications to this method were also examined, and no improvements were observed. The optimal excystation protocol for E. krijgsmanni was elucidated as of now.

Increasing antibiotic resistance in Mycoplasma hyopneumoniae (Mhp) necessitates alternative treatments. Piper sarmentosum extracts (PSE) and guava extracts (GE) show promise due to their antimicrobial and antioxidant properties. This study evaluates the combined in vitro effects of PSE and GE against Mhp and their antioxidant activities. Anti-Mhp activity was assessed using microdilution minimum inhibitory concentration (MIC) and synergy via fractional inhibitory concentration index (FICI), while antioxidant capacity was measured with 2,2’-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and ferric reducing antioxidant power (FRAP) assays for individual extracts, and a 1:1 mixture (based on the results of FICI). Phytochemical profiling used high performance liquid chromatography (HPLC) to quantify main six bioactive compounds. HPLC showed PSE contained vitexin-2-O-rhamnoside (8.58 mg/g) and pellitorine (3.31 mg/g); GE had quercetin-3-O-glucuronide (6.85 mg/g), quercetin (3.72 mg/g), isoquercitrin (0.51 mg/g), and avicularin (1.37 mg/g). PSE (MIC 0.78 mg/mL) and GE (MIC 0.39 mg/mL) exhibited anti-Mhp effects, with their 1:1 combination being additive (FICI=0.75) and reducing MIC to 0.195 mg/mL. This mix also enhanced antioxidant activity by 9.0% in ABTS and 27.0% in FRAP assays. The 1:1 combination of PSE and GE demonstrated superior anti-Mhp activity (additive effect) and enhanced antioxidant properties (synergistic effect) compared to the individual extracts. These findings highlight the potential of these extracts as phytopharmaceutical agents.

Prolonged lung inflammation leads to the development of asthma and approximately 27.8% of adult patients with asthma suffer from depression. We examined the effect of the prostaglandin F_2α (PGF_2α) receptor (FP receptor) agonist, fluprostenol, on ovalbumin (OVA)-induced asthma and asthma-related depression in mice. Repeated fluprostenol+OVA administration increased OVA-induced inflammatory cell infiltration and mRNA expression of inflammatory mediators in the lung. In contrast, in the tail suspension and forced swim tests, fluprostenol+OVA administration significantly reduced the immobile time compared with saline+OVA-administered mice. Fluprostenol+OVA treatment significantly upregulated serotonin 1A receptor and tryptophan hydroxylase in the hippocampus compared with the expression in saline+OVA mice. These results suggest that PGF receptor (FP receptor) stimulation promotes lung inflammation but attenuates depression, possibly via the serotonin pathway.

Four genetically identical cloned dogs were produced from a single donor using the somatic cell nuclear transfer technique. They stayed in the same facility during youth period, and were later assigned to different locations according to their roles as working dogs. In this study, we report observations from four genetically identical cloned working dogs exposed to different environments, including an airport and a police station, for two years. Hair and fecal samples were collected to assess potential environmental influences on mineral content and intestinal microbiota composition. The concentrations of toxic minerals, such as Pb and Ni, were significantly higher in the hair mineral analysis of the dog exposed to the airport compared to those of the three police station-working dogs. Fecal microbiota analysis showed that the intestinal microbiota of the airport-exposed dog was simplified, with an increased proportion of Actinobacteria and a decreased proportion of Lachnospiraceae and Fusobacteria. Our observations suggest that environmental conditions may influence the accumulation of harmful substances in the body and affect intestinal microbiota community. This finding also highlights the potential of non-invasive methods for evaluating workplace-related environmental influences on working dogs. These results suggest that different environments may contribute to physiological differences even among genetically identical individuals. Future studies should investigate the mechanisms underlying these variations and assess their long-term implications for canine health monitoring with larger sample sizes.

Contamination of cell lines with bovine viral diarrhea virus (BVDV) derived from fetal bovine serum (FBS) is a major obstacle in cell biology experiments. Although the selection of BVDV-free FBS and constant viral checking are important to prevent the viral contamination of cells, they have been ineffective in the past contamination of several cell lines available from cell banks. Recently, a novel method to eliminate BVDV from one contaminated cell line, LFBK-α_vβ₆ cells, using BVDV inhibitor DB772 was reported. To confirm the reproducibility and possibility of practical use of this method in other cell lines, we attempted BVDV elimination using DB772 from five contaminated cell lines: BL3.1, EBTr, A26, FLK-BLV, and CC81. Treatment with DB772 significantly decreased viral genome counts in all BVDV-contaminated cells; however, cytotoxicity-like growth suppression was observed, in contrast to previously reported findings. Moreover, continuous cell passages in the presence of DB772, conducted in imitation of a previous study, resulted in the failure of elimination with rebound of viral genomes after several cell passages. Hence, we modified the method to combine limiting dilution of cultured cells with drug treatment and finally succeeded in BVDV elimination in four of the 5 cell lines, except for EBTr cells, which exhibited strong suppression of cell proliferation after DB772 treatment. Taken together, our findings will help to solve the problem of BVDV contamination in cell lines.

Equine coronavirus (ECoV) causes fever, anorexia and sometimes enteric signs (e.g. diarrhea) in adult horses. Three ECoV outbreaks among draft horses at the same Japanese racecourse have been reported previously, in 2004, 2009 and 2012. We describe the epidemiological, clinical and genetic features of the fourth ECoV outbreak at the same racecourse in 2025. In January 2025, a sudden increase in the number of draft horses showing various combinations of fever, anorexia and enteric signs was observed at Obihiro Racecourse, in Hokkaido, Japan. Of about 800 horses, 176 showed clinical signs. Specifically, 83.5% (147/176) horses had fever, 98.9% (174/176) were anorexic, and 10.2% (18/176) had enteric signs. Real-time reverse transcription –polymerase chain reaction detected ECoV RNA in 23 of 26 horses. Seroconversion to ECoV was confirmed in 25 of 26 horses by neutralization assay. The results indicate that all 26 horses tested were infected with ECoV. Molecular analyses showed that the spike and nucleocapsid genes were highly conserved compared to other ECoVs. However, phylogenetic analysis showed that these genes of the 2025 virus did not form a group with the strains detected in the previous three outbreaks. These results indicate that ECoV was the cause of the sudden increase in the number of draft horses with fever, anorexia and enteric signs. Although the origin of the 2025 virus is unknown, this outbreak was likely caused by a virus different from the one that caused the previous outbreaks.

Bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) are retroviruses infecting cattle, with BLV being a known cause of enzootic bovine leukosis. In contrast, BIV is considered non-pathogenic and has been less studied, despite its relevance to animal lentiviral evolution. This study aimed to develop and validate a multiplex real-time PCR assay for simultaneous detection of BIV proviral DNA and quantification of BLV proviral load. Novel primers and probes targeting the BIV pol gene were designed. The assay demonstrated a detection limit of 10 copies per reaction with a probability of 83%, comparable to conventional nested PCR. Importantly, the addition of BIV primers and probes did not interfere with the accurate quantification of BLV. Furthermore, BIV detection was unaffected even in samples with high BLV proviral loads, confirming the assay’s robustness for simultaneous detection. The validated assay was applied to a molecular epidemiological survey of 6,051 cattle in the Kyushu region of Japan, conducted between 2021 and 2025. No BIV proviral DNA was detected in any sample, indicating a low prevalence of BIV in this population. This multiplex PCR assay provides a sensitive, efficient, and labor-saving tool for integrated surveillance of retroviral infections in cattle. Furthermore, it offers a valuable platform for future studies on the epidemiology and ecological role of BIV in bovine populations.

Avian haemosporidian parasites cause high pathogenicity and lethal infections to non-native or introduced birds. Crested ibis (Nipponia nippon) resulted extinct in Japan but the Chinese population was introduced to Japan. Understanding the prevalence of pathogens of rare species is important for their conservation but it is often difficult to investigate them due to the limitation of the access. We collected parasite vector insects in 2022 and 2023 around the crested ibis conservation center in Sado Island. Among collected 214 mosquitoes and 585 biting midges, three Haemoproteus genetic lineages were detected from four Culicoides biting midges for the first time. Our findings suggested that the transmission of Haemoproteus could occur between biting midges and wild birds including crested ibis.