Typical skeletal muscles are composed of mixed muscle fiber types, which are classified as slow-twitch (type I) and fast-twitch (type II) fibers, whereas pectoralis major muscles (PMs) in broiler chickens are 100% composed of type IIb fast-twitch fibers. Since metabolic properties differ among muscle fiber types, the combination of muscle fiber types is involved in physiological functions and pathological conditions in skeletal muscles. In this study, using serial block-face scanning electron microscopy, we compared three-dimensional (3D) mitochondrial properties in type IIb fibers in broiler PMs and those in type I fibers of broiler gastrocnemius muscles (GMs) heterogeneously composed of slow- and fast-twitch muscle fibers. In type I fibers in the GMs, elongated mitochondria with numerous interconnections to form a substantial network among myofibrils were observed. Along with lipid droplets sandwiched by mitochondria, these features are an adaptation to effective oxidative respiration and constant oxidative damage in slow-twitch muscle fibers. In contrast, type IIb fibers in the PMs showed small and ellipsoid-shaped mitochondria with few interconnections and no lipid droplets, forming a sparse network. The mitochondrial spatial network comprises of active mitochondrial dynamics to reduce mitochondrial damage; therefore, type IIb fibers possess physiologically low capacity to maintain mitochondrial wellness due to static mitochondrial dynamics. Based on 3D mitochondrial properties, we discussed the contrasting physiological functions between type I and IIb fibers and proposed a high contractile power and low stress resistance as unique physiological properties of broiler PMs.

Porcine edema disease (ED) is a life-threatening toxemia caused by enteric infection with Shiga toxin 2e (Stx2e)-producing Escherichia coli (STEC) in weaned piglets. We previously reported that the stx2eB-transgenic lettuce 2BH strain shows potential for use as an oral vaccine candidate against ED. However, the 2BH strain expressed a hemagglutinin (HA)-tag together with Stx2eB and contained non-canonical N-glycosylation. Therefore, we developed two Stx2eB-lettuce strains, the 3 (G+) strain in which the HA-tag was removed from 2BH, and the 3 (G-) lettuce strain, in which the 73rd Asn was replaced with Ser to prevent non-canonical N-glycosylation of Stx2eB from the 3 (G+) strain. We examined the protective effect of these newly developed two strains compared with the previous 2BH strain against ED using a colostrum-deprived piglet STEC infection model. We found that the N-glycosylated 2BH and 3 (G+) strains relieved the pathogenic symptoms of ED in STEC-challenged piglets, whereas the non-glycosylated 3 (G-) strain did not. N-Glycosylation of the Stx2eB product in lettuce may be involved in the immune response in piglets.

When a large-scale epidemic of porcine epidemic diarrhea (PED) occurred in 2013 in Japan, feedback feeding (feeding feces and gut tissues of infected piglets) was attempted to impart immunity to sows and immunize nursing piglets via breastfeeding. This study evaluated the effect of feedback feeding on PED control at 172 farms in Kagoshima and Miyazaki Prefectures. Univariable and multivariable generalized linear models were used to analyze the associations between conduct of feedback feeding and damage from the outbreak (outbreak period and the number of piglet deaths) at the farm level. The within-farm outbreak period shortened over time after the regional outbreak began on Kyushu Island (P=0.009) and was longer on large-scale farms (mean 66.0 days, P=0.003) than small-scale farms (29.4 days) and on farms that used feedback feeding (145.2 days, P=0.059) than those that did not (66.0 days). The number of dead piglets decreased over time since the first regional case (P<0.001) and was higher at farrow-to-finish farms (3.8 piglets/sow, P<0.001) than reproduction farms (0.7 piglets/sow). The effect of feedback feeding on the number of dead piglets was not significant, but its interaction term with farm style had a significant effect (5.0 more piglet deaths at reproduction farms than fallow-to-finish farms, P=0.001). These results suggest that feedback feeding made the damage from PED worse, though it was well established at a later stage of the regional PED epidemic.

The aim of this study was to evaluate the microbiota of normal milk in dairy cows and their relationship with host factors, such as the age of the cow (Age), somatic cell counts in milk (SCCs), and days in milk (DIM). We investigated 48 milk samples from 22 cows with no systemic or local clinical signs using MinION^TM nanopore sequencing for a 16S rRNA gene amplicon. Bacterial richness was positively correlated with the DIM (P=0.043), and both the Shannon-Wiener Index and Simpson’s Index, which are metrics of alpha-diversity, were also significantly positively correlated with the SCC (P<0.001). The composition ratios of both Actinobacteria at the phylum level and Kocuria spp. at the genus level in the milk microbiota were significantly correlated with the SCC (P<0.001 and P<0.001, respectively). In the beta-diversity test, the one-way analysis of similarities test showed a significant difference (P=0.0051) between the low- and high-SCC groups. This study clarified that the composition of the normal milk microbiota in this herd was related to the SCC. It also raised the possibility of variations in bacterial genera in the normal milk microbiota between the low- and high-SCC groups. However, to clarify the actual condition of the milk microbiota and to elucidate the relationship with the SCC, it is necessary to perform further analyses taking into account not only the relative abundance, but also the absolute abundance of microbes.

The accurate identification of mastitis‐causing bacteria assists in effective management by both dairy farmers and veterinarians and can be used to implement the selective use of antimicrobials for treatment. The purpose of this study was to evaluate the ability of our developed anti–ribosomal protein-L7/L12 antibody–coated immunochromatographic strip (ICS) test to detect coliforms in milk by comparing the results with the bacteriological culture method. We investigated the performance of the ICS test as compared with the bacteriological culture method using 308 milk samples from clinical bovine mastitis. First, to determine the optimal ICS test cutoff point for detecting coliform mastitis, we developed a receiver-operating characteristic curve. The result showed that the cutoff point was at 0.5 of our index. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value of the ICS test were 81.3%, 84.8%, 69.2%, and 91.54%, respectively. As the clinical signs increased in severity, the F-measure, a weighted harmonic mean of the sensitivity and overall PPV performance, increased. Because it is especially important to treat clinical mastitis appropriately in the early stages of detection, the ICS test, which can be used by both dairy farmers and veterinarians on dairy farms, is considered to be a useful tool for detecting coliform mastitis, which often presents with severe signs.

We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2–16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.

An observational study was conducted to comparatively assess the efficacy of three different oral rehydration and buffering solutions, differentiated by their strong ion difference (SID) concentration, for treatment of neonatal calves with naturally acquired diarrhea. The SID concentrations tested were 100 mM, 170 mM and 230 mM for treatments SID₁₀₀, SID₁₇₀ and SID₂₃₀, respectively. Clinical assessment and blood gas analysis were completed for 18 diarrheic calves once pre- and twice post- (6 and 24 hr after) oral administration with one of the three treatments. A repeated measure mixed model approach was used to analyze (a) the within-group efficacy of each treatment over time and (b) the between-group comparison at each timepoint. SID₂₃₀ treatment resulted in a significant increase in blood pH, HCO₃⁻, BE, SID and Na⁺ at 6 and 24 hr after treatment, and a significant decrease in AG and K⁺ by 24 hr after treatment. There were no significant changes in any of the blood gas parameters after treatment with SID₁₀₀ and SID₁₇₀. SID₂₃₀ treatment also resulted in blood gas parameter changes that were significantly different to the other two groups. These results suggest that the optimum SID concentration for the treatment of calves with diarrhea is likely to be higher than current recommendations.

A 4-year, 7-month-old Holstein cow presented with anorexia. Physical examination revealed masses in the interscapular region and vagina. Blast cells were detected in the masses and peripheral blood by fine needle aspiration cytology and hematological examination. By bone marrow aspiration, blast cells constituted up to 24.2% of all nucleated cells, and 22% and 2% of non-erythroid cells stained positive for myeloperoxidase and alpha-naphthyl acetate esterase (ANAE), respectively. Pathological examination revealed the mass lesions consisted of a proliferation of tumor cells, which were positive for monocytic markers (HLA-DR and Iba-1). The cow was diagnosed with acute myelomonocytic leukemia (AMML). Even when tumor cells are ANAE-negative, AMML cannot be completely ruled out and should be considered when diagnosing cattle with leukemia/lymphoma.

The aim of this study was to evaluate the impacts of ophthalmic findings obtained from both macroscopic examination and ocular ultrasonography when diagnosing bovine endophthalmitis. A newborn crossbreed (Japanese black and Holstein breeds) calf was suspected of visual impairment and central nervous system (CNS) symptoms, such as decreased activity and weak drinking performance. This calf was found to display macroscopic signs, such as clouded lens, convergent strabismus, and horizontal nystagmus, in both eyes. On ocular ultrasonography of both eyes, a V-shaped, thickened, hyperechoic structure was present in the anechoic vitreous humors, indicating retinal detachment. The animal died 4 days after the examination. Sepsis was evident in this case, as Escherichia coli was isolated from multiple organs. The autopsy and histological examination revealed meningitis, encephalitis, and secondary hydrocephalus in the CNS, and endophthalmitis and retinal detachment in both eyes. In this case, the ophthalmic findings did not provide definitive evidence for a diagnosis of endophthalmitis. However, this study indicated that retinal detachment might be an ultrasonographic finding that is suggestive of bovine endophthalmitis.

We investigated the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among dogs in the Tokyo area via enzyme-linked immunosorbent assay (ELISA) using the spike protein as the target antigen. Plasma samples from 494 household dogs and blood-donor dogs were tested from July 2020 to January 2021. Of these samples, three showed optical densities that were higher than the mean plus two standard deviations of the mean of the negative-control optical densities (ODs). Of these three samples, only the sample with the highest OD by ELISA was confirmed positive by virus neutralization testing. The positive dog presented no SARS-CoV-2-related symptoms. The positivity rate of SARS-CoV-2 infections among dogs in the Tokyo area was approximately 0.2%.

The ideal treatment for intracranial histiocytic sarcoma (HS) remains unclear. Herein, we report a case of intracranial HS that was successfully treated using prednisolone and radiation therapy. The patient was a 9-year-old spayed female Pembroke Welsh Corgi that presented with epileptic seizures. Magnetic resonance imaging revealed a contrast-enhancing mass adjacent to the right piriform lobe. Prednisolone administration (1 mg/kg/day) decreased the lesion size. Additional palliative radiation therapy (total dose, 37 Gy) resulted in complete disappearance of the lesion. However, on day 164, the dog’s neurological signs deteriorated, and she was euthanized. Necropsy revealed an intracranial metastasis of HS via the cerebrospinal fluid without any extracranial metastasis. Nonetheless, combined prednisolone and radiation therapy might be effective in treating intracranial HS.

Investigation of the role of animals that have recovered and survived from African swine fever (ASF) in carrying the ASF virus is currently intense and ongoing. However, no clear definition of the carrier stage has been established. The aim of the present study was to establish criteria to elucidate a clear status of survival in naturally ASF-infected domestic pigs in Vietnam. Seroconversion from previous infection was confirmed by serological assay, and the absence of the viral genome in various organs was also assured by molecular analysis of a partial p72 gene. We recognized that histopathological evidence could benefit from further insights into the status and role of the surviving animals; therefore, we performed a histopathological study on four pigs from farms with a history of ASF outbreak. We found fibrotic changes in the reparative process as the main finding in all four pigs. Immunohistochemical detection of viral protein revealed an interesting result. Despite the negative result from viral genome detection, the p30 protein gave a positive signal in the tonsils, lung, and stomach. This raises the possibility of stress-induced viral reactivation in long-term survivors and the risk of further outbreaks from human handling of contaminated carcasses.

An eighteen-year-old female Eurasian otter became emaciated and died. Necropsy examination revealed nose and thoracic cutaneous masses, abdominal subcutaneous mass, and multiple nodules in the liver and lungs. Malignant melanoma was found in the nose cutaneous mass and to have metastasized to the liver, lungs, kidneys, adrenal glands, mammary glands and left mandibular lymph node. The neoplastic cells were labeled for vimentin, melanoma, and S100. The cutaneous mass in the thoracic area consisted of spindle shaped neoplastic epithelial cells and was diagnosed as trichoblastoma. Mammary gland adenoma was observed in the abdominal subcutaneous mass. This is the first report of primary three neoplasms of malignant melanoma, trichoblastoma and mammary gland adenoma in a Eurasian otter.

In July 2020, a sow in a breeding herd in the Chiba Prefecture, Japan, suffered abortion. A necropsy revealed pale pulmonary foci scattered in the two fetuses. Histologically, multifocal pulmonary necrosis was detected with numerous yeasts. The yeast was positively stained using the periodic acid-Schiff reaction and Grocott’s silver stain. Molecular identification indicated that the yeast was Candida parapsilosis. In conclusion, our results suggested that C. parapsilosis caused multifocal necrotizing pneumonia in the two fetuses. This study is the first report of a swine abortion with C. parapsilosis infection.

Skin lesions possibly caused by Papillomavirus infections in two four-toed hedgehogs are described. In case 1, there was a papillary mass on the right hind limb. Histologically, the mass was consistent with a viral papilloma. In the other case, multifocal papillary masses with erosions and ulcers were found throughout the body, mainly on the extremities. Histology showed continuative lesions composed of acanthosis, Bowenoid in situ carcinoma, and squamous cell carcinoma, with abrupt transitions between the lesions. In both cases, keratinocytes in the granular layer infrequently had features of koilocytes and intranuclear inclusion bodies, and immunohistochemical staining was positive for anti-human papillomavirus antibody. To the best of the authors’ knowledge, this is the first pathological documentation of possibly papillomavirus-associated skin lesions in four-toed hedgehogs.

Microparticles (MPs) are released from budding plasma membranes into body fluids. The use of flow cytometry for the measurement of MP in canines has not been standardized. In this fundamental study, we compared the effect of anticoagulant agents, such as acid-citrate-dextrose (ACD) and heparin on the measurement of canine MPs in platelet-free plasma (PFP) using flow cytometry. In addition, we used annexin V, carboxyfluorescein succinimidyl ester (CFSE), or calcein tetraacetoxymethyl ester (calcein-AM), and explored the characteristics of the staining reagents in MP detection using flow cytometry. We were able to measure canine MPs in PFP prepared from ACD-anticoagulated blood using flow cytometry, in which the highest positive rate for fluorescent staining was observed when CFSE was used.

From 2012 to 2021, prevalence of pathogenic Yersinia in wild rodents captured in Fukushima Prefecture, Japan was investigated twice a year to clarify the ecology of this pathogen in wild rodent populations. Pathogenic Yersinia enterocolitica O8 was isolated from 13 (1.7%) of 755 wild rodents. The Y. enterocolitica O8 isolates harbored three virulent genes (ail, fyuA, and virF). This pathogen was isolated repeatedly from wild rodents in April 2015, 2016, and 2017, in June and November 2020, and in April 2021, which was 6 of 19 times of observations. All Y. enterocolitica O8 isolates showed the same PFGE patterns. These results indicated that the same clone of pathogenic Y. enterocolitica O8 has been maintained in wild rodent populations in Fukushima Prefecture. Therefore, wild rodent populations contribute substantially to the continuous transmission of Y. enterocolitica O8 and its persistence in the ecosystem. This is the first report on the isolation of pathogenic Y. enterocolitica O8 in wild rodents in Fukushima Prefecture, Japan.

Polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) is commonly used for genotyping bovine leukemia virus (BLV) in slaughterhouses. However, unclassified BLV genotypes have been sporadically reported. To assess the current status of BLV genetic characterization in cattle, PCR-RFLP was performed on blood samples of 170 cattle (84 Japanese Black, 60 Japanese Black x Holstein, and 26 Holstein) from 17 farms (5 prefectures) at a slaughterhouse in Aichi Prefecture in 2019. A total of 65 samples (38.2%) were BLV positive, and genotype 1 was the most predominant (56/65 samples), followed by genotypes 3 (6 samples) and 5 (1 sample), and two unclassified samples. No relationship between the genotypes and breeds was observed. Sequence and phylogenetic analyses demonstrated that unclassified BLV genotypes clustered with genotype 1 sequences were, therefore, not new genotypes.

There has been no report on Chronic wasting disease (CWD) cases in Japan to date; however, there is concern about the geographic spread of CWD. To clarify the CWD status in Japan, we conducted CWD monitoring using real-time quaking-induced conversion (RT-QuIC) assay which can detect the low level of CWD prions. A total of 690 obex samples collected from sika deer and Reeves’s muntjac in Hokkaido and Honshu was tested for CWD prions. No CWD-positive cases were found, suggesting that CWD is nonexistent in Japan. Our results also indicate that RT-QuIC assay is useful for continuous monitoring of CWD. Furthermore, nucleotide sequence analysis of the PrP gene revealed sika deer in Japan harbor CWD susceptible allele.

A 3-year-old male domestic shorthair cat presented with sudden ataxia. Neurologic examination showed complete loss of proprioception in the thoracic and pelvic limbs. Computed tomography and magnetic resonance imaging revealed a non-metallic foreign body penetrating the spinal cord. The foreign body was removed by the ventral approach to the atlanto-occipital junction. Mild improvement of proprioception was observed the day after surgery. In a follow-up two months after surgery, the owner reported a complete recovery of the patient, showing a normal gait. To the author’s knowledge, this is the first case report describing successful removal of an intramedullary foreign body penetrating cervical spinal cord by ventral approach in a cat.

Canine gastroesophageal intussusception (GEI) is a rare and life-threatening condition that requires prompt diagnosis and treatment. A 19-day-old Siberian Husky with a 4-day history of regurgitation was diagnosed with GEI based on the findings of computed tomography (CT) performed without anesthesia. Endoscopic reduction of intussusception was impossible; thus, surgical reduction by traction of the duodenum was performed. CT revealed improvement of megaesophagus 82 days postoperatively. Eleven months postoperatively, fluoroscopy showed recovery to nearly normal esophageal motility. Two years postoperatively, no clinical signs were reported. CT is useful to diagnose GEI in neonate puppies with poor abdominal fat and to assess the gastric edema and the anatomical association of stomach with other organs. Fluoroscopy is helpful for evaluating postoperative esophageal motility.

The shape of the white line of the hoof is closely related to the shape of the notch on the dorsal distal bearing border of the distal phalanx (P3). In this study, a radiographic survey of the P3 of both forelimbs of 163 Thoroughbred yearling horses was conducted. The correlation of the depth and width of the notch were analyzed with the toe white line separation grades (0 to 3). As a result, the toe white line separation grade increased, the depth and the width of the notch also increased significantly. Radiographic examination of the P3 of the forelimbs might be useful for deciding whether to implement hoof care to prevent onset of toe white line separation.

Podoplanin is expressed in various human tumors where it promotes tumor progression, epithelial-mesenchymal transition, and distant metastasis. Podoplanin is also expressed in cancer-associated fibroblasts and induces tumor malignancy. The objective of this study was to evaluate podoplanin expression in various types of feline tumor tissues. Immunohistochemical analysis revealed that podoplanin was expressed in cells of 13/15 (87%) squamous cell carcinomas and 5/19 (26%) fibrosarcomas. Moreover, cancer-associated fibroblasts expressed podoplanin in most tumor types, including 18/21 (86%) mammary adenocarcinoma tissues. Our findings demonstrate that various types of feline tumor tissues expressed podoplanin, indicating the importance of the comparative aspects of podoplanin expression, which may be used as a novel research model for podoplanin biology.

Lead (Pb) exposure occurs together with other metals including zinc (Zn). This study investigated the impact of Zn on Pb tissue accumulation and Pb-induced toxicities. Animals (n=6 rats per group) were exposed to lead acetate (PbAc) or a combination of PbAc and zinc acetate (ZnAc) under the following groups: control (deionized water), low PbAc [12 mg/kg PbAc (3 mg PbAc/rat/day)], low PbAc–ZnAc [12 mg/kg PbAc (3 mg PbAc/rat/day) + 0.2 mg ZnAc/rat/48 hr], high PbAc [120 mg/kg (30 mg PbAc/rat/day)], and high PbAc–ZnAc [120 mg/kg (30 mg PbAc/rat/day) + 1 mg ZnAc/rat/48 hr] for 8 weeks. A significant reduction in body weight gain was observed in the high PbAc group relative to the control group. Muscles and testes both had reduced and increased Pb uptake in low PbAc–ZnAc and high PbAc–ZnAc groups compared to PbAc only groups, respectively. Bone Pb levels in the high PbAc–ZnAc group were lower than the high PbAc group. Zinc co-administration attenuated Pb-induced inhibition of delta aminolaevulinic acid dehydratase enzyme and enhanced catalase enzyme activity at a high level of exposure. Moreover, ZnAc seems to have minimized the effects of Pb-induced mRNA dysregulation in antioxidant and antiapoptotic enzymes encoding genes. Heme oxygenase-1 was downregulated in the kidney and brain in the low PbAc group. Liver glutathione peroxidase and thioredoxin reductase-1 were downregulated in the high PbAc group. These findings suggest that zinc co-administration with lead may partially mitigate against Pb-induced toxicities.

Infectious bursal disease (IBD) causes severe economic damage to the poultry industry worldwide. To prevent IBD virus (IBDV) infection, live virus vaccines have been widely used in chickens having wide-ranging levels of maternally derived antibodies. But, the risks of infection with other pathogens because of lesions related to atrophy of the bursa of Fabricius in vaccinated chickens are a concern. To resolve the problems, a recombinant turkey herpesvirus (HVT) vaccine expressing IBDV-VP2 protein (rHVT-IBD) has been developed. However, the induction of neutralizing antibodies by rHVT-IBD against a virulent IBDV might be delayed compared with that by the live IBD vaccine, leading to the high risks of IBDV infection for young chickens. To find the best selection of IBDV vaccine for the onset of immunity, we examine the protective efficacy of a novel in ovo-attenuated live IBDV (IBD-CA) vaccine and the rHVT-IBD vaccine in young chickens challenged with a very virulent IBDV (vvIBDV) strain. We show that the protective efficacy of IBD-CA vaccine was higher than that of the rHVT-IBD vaccine in 14-day-old chickens challenged with the vvIBDV strain, leading to the risk of IBDV infection for young chickens when vaccinated with rHVT-IBD. Our results suggest that farmers should select the best vaccines to maximize vaccine efficacy in consideration of the vaccine characteristics, prevalence levels of IBDV in the areas, and initial MDA levels of the chickens since the attenuated live and recombinant vaccines play a role in the different vaccine efficacies.

Sporadic spreads of swine-origin influenza H3N2 variant (H3N2v) viruses were reported in humans, resulting in 437 human infections between 2011 and 2021 in the USA. Thus, an effective vaccine is needed to better control a potential pandemic for these antigenically distinct viruses from seasonal influenza. In this study, a candidate vaccine strain with efficient growth capacity in chicken embryos was established through serial blind passaging of A/Indiana/08/2011 (H3N2)v in mice and chicken embryos. Seven amino acid substitutions (M21I in PA; A138T, N165K, and V226A in HA; S312L in NP; T167I in M1; G62A in NS1 proteins) were found in the passaged viruses without a major change in the antigenicity. This mouse- and egg-adapted virus was used as a vaccine and challenge strain in mice to evaluate the efficacy of the H3N2v vaccine in different doses. Antibodies with high neutralizing titers were induced in mice immunized with 100 µg of inactivated whole-virus particles, and those mice were significantly protected from the challenge of homologous strain. The findings indicated that the established strain in the study was useful for vaccine study in mouse models.

Efficacy of African swine fever virus (ASFV) inactivation using five commercially supply compound disinfectants was evaluated under various condition. Virucidal efficacy demonstrated that products A and E could inactivate at 1:800 within 1 min for both temperatures, while products B, C and D inactivated at 1:400. However, product D could inactivate at 1:800 when the exposure time was extended to 30 min and effected only 20°C. In addition, the cytotoxicity demonstrated that products A, B, C, D and E did not significantly affect to cell at 1:51,200, 1:12,800, 1:12,800, 1:25,600 and 1:12,800 dilution, respectively. In conclusion, these disinfectants could inactivate ASFV, however, the application of these products should be performed under safety precautions to prevent cytotoxicity in humans and animals.

Tuberculosis (TB) is a chronic inflammatory and zoonotic disease caused by Mycobacterium tuberculosis complex (MTBC) members, affecting several domestic animals, wildlife species and humans. The preliminary investigation was aimed to detect antibody against MTBC among indigenous wildlife which are free-ranged wild boar, free-ranged wild macaques and captive Asian elephants in selected areas of Selangor and elephant conservation centre in Pahang, respectively. The results indicate that MTBC serodetection rate in wild boar was 16.7% (7.3–33.5 at 95% confidence interval (CI)) using an in-house ELISA bPPD IgG and 10% (3.5–25.6 at 95% CI) by DPP^®VetTB assay, while the wild macaques and Asian elephant were seronegative. The univariate analysis indicates no statistically significant difference in risk factors for sex and age of wild boar but there was a significant positive correlation (P<0.05) between bovine TB in dairy cattle and wild boar seropositivity in the Sepang district.

The screening of reference genes for real-time quantitative PCR (qPCR) in forest musk deer (FMD) tissue is of great significance to the basic research on FMD. However, there are few reports on the stability analysis of FMD reference genes so far. In this study, We used qPCR to detect the expression levels of 11 reference gene candidates (18S rRNA, beta-actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box-binding protein [TBP], hypoxanthine phosphoribosyltransferase 1 [HPRT1], tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide [YWHAZ], hydroxymethylbilane synthase [HMBS], eukaryotic translation elongation factor 1 alpha 1 [EEF1A1], succinate dehydrogenase complex flavoprotein subunit A [SDHA], peptidylprolyl isomerase B [PPIB], and ubiquitin C [UBC]) in heart, liver, spleen, lung and kidney of FMD. After removing 18S rRNA on account of its high expression level, geNorm, NormFinder, BestKeeper and ΔCt algorithms were used to evaluate the expression stability of the remaining genes in the five organs, and further comprehensive ranking was calculated by RefFinder. According to the results, the selected reference genes with the most stable expression in the heart of FMD are SDHA and YWHAZ, while in the liver are ACTB and SDHA; in the spleen and lung are YWHAZ and HPRT1; in the kidney are YWHAZ and PPIB. The use of common reference genes in all five organs is not recommended. The analyses showed that tissue is an important variability factor in genes expression stability. Meanwhile, the result can be used as a reference for the selection of reference genes for qPCR in further study.

A racing pigeon (Columba livia var. domestica), a straggler from Taiwan, was sheltered in Nara Prefecture, Japan in 2020. This pigeon showed hemolysis and elevated levels of hepatobiliary and muscle enzymes. Gametocytes of Haemoproteus columbae (Apicomplexa: Haemosporida) were observed within the host erythrocytes in thin blood smears. A partial sequence of the mitochondrial cytochrome b gene amplified from blood DNA was identical to the lineage HAECOL1 previously reported from pigeons worldwide. This is the first record of H. columbae infection in a sheltered bird in Japan.