Biotin is a vital coenzyme involved in diverse metabolic pathways and plays a key role in hoof health by supporting keratin synthesis and the protective barrier of the hoof. This study systematically investigated the effects of rumen-protected biotin (RPB) on hepatic metabolic networks, redox homeostasis, and hoof health in lactating dairy cows using an integrated multiomics approach. The RPB supplement consisted of 2.1% biotin (purity ≥99%), 62.9% glucose, and 35% hydrogenated palm oil fatty acids. Eighty multiparous Holstein cows were stratified by parity (2.48 ± 0.69), body weight (637.76 ± 55.71 kg), body condition score (3.01 ± 0.20), days in milk (141 ± 16), and average milk yield (32.35 ± 4.36 kg/d). They were assigned for 75 d using a randomized block design to 4 treatment groups: Control (Con; basal diet, n = 20), low-RPB diet (LRPB; 0.5 g/d RPB, n = 20), mid-RPB diet (MRPB; 1.0 g/d RPB, n = 20), or high-RPB diet (HRPB; 2.0 g/d RPB, n = 20). Milk and blood samples were collected on d 0, 15, 30, 45, 60, and 75 for analysis of milk composition and serum biochemical parameters. Data were analyzed using mixed models with orthogonal polynomial contrasts to evaluate linear and quadratic effects of RPB. Among the markers of liver function, serum albumin increased but total cholesterol, alanine aminotransferase, aspartate aminotransferase, and total bilirubin decreased in a linear and quadratic fashion with higher doses of RPB. Feeding RPB increased serum total antioxidant capacity, glutathione, and superoxide dismutase in a linear and quadratic fashion, while malondialdehyde decreased. Incremental feeding of RPB decreased linearly the serum type II collagen C-terminal peptide concentration and lameness scores. Further, serum cartilage oligomeric matrix protein concentration decreased in a linear and quadratic fashion, whereas serum procollagen IIA N-terminal propeptide and hoof horn hardness increased in a linear and quadratic fashion. Based on serum biochemical and hoof health results on d 75, the Con and MRPB groups were selected for proteomic and metabolomic analyses of serum. Proteomics revealed that RPB upregulated key proteins involved in antioxidant reactions and keratinization, including GSR, GCLC, GPX3, TGM1, and TGM3. Metabolomics identified l-cysteine, glycine, and pyruvate as key metabolites associated with RPB suggesting upregulated glutathione synthesis and flux through the tricarboxylic acid cycle. Integrated proteomics and metabolomics analyses revealed that GSR, GCLC, GPX3, TGM1, and TGM3 were positively correlated with l-cysteine and glycine but negatively correlated with γ-glutamylcysteine and palmitic acid. Overall, feeding RPB reduces oxidative stress and improves liver function in part by enhancing glutathione metabolism while reducing lipid peroxidation. Further, RPB promotes keratinization and limits cartilage degradation, thereby enhancing hoof health. These responses to dietary RPB supplementation provide molecular evidence for its targeted application in dairy herd nutrition management.

Fusobacterium necrophorum (F. necrophorum), a major pathogen responsible for foot rot in dairy cows, causes interdigital skin ulceration, sole necrosis, and hoof wall degeneration or detachment. Emodin (EMO), a naturally occurring anthraquinone derivative, exhibits anti-inflammatory, antimicrobial, and antitumor properties. However, whether it can alleviate F. necrophorum-induced pyroptosis in bovine digital fibroblasts (BDFs) and the underlying mechanisms remains unexplored. This study aimed to investigate whether EMO participates in the regulation of F. necrophorum-induced pyroptosis in BDFs by modulating the NOD-like receptor protein 3 (NLRP3)/gasdermin D (GSDMD) signaling pathway. In vitro experiments demonstrated that it exhibits significant antibacterial activity against F. necrophorum, with a minimum inhibitory concentration (MIC) of 128 μg/mL achieving an 88.2% inhibition rate (P < 0.001). The BDFs infection model was established using F. necrophorum (MOI 75) for 12 h, and pretreatment with 50 μM EMO significantly alleviated the inhibitory effects of F. necrophorum on the viability and migratory capacity of BDFs (P < 0.001). In a skin explant model, treatment with EMO reduced inflammatory cell infiltration, mitigated collagen fiber disorganization, and alleviated cellular changes including membrane rupture, endoplasmic reticulum dilation, ribosome detachment, mitochondrial swelling, and blurred cristae structure of the mitochondria. Moreover, EMO alleviated pyroptosis induced by F. necrophorum and NLRP3 agonist. It also downregulated NLRP3 expression and consequently inhibited the protein level expression of the pyroptosis markers, including GSDMD, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 (P < 0.001). These results support the theoretical basis for using EMO to regulate F. necrophorum-induced pyroptosis via the NLRP3/GSDMD pathway and provide insight for controlling foot rot in dairy cows.

Inactive ovaries (IO) are a common reproductive disorder in early lactating dairy cows. This condition significantly reduces reproductive efficiency and economic returns. Although many studies have explored the metabolic characteristics of postpartum IO, systematic proteomic analyses using multiple biological samples are still lacking. This study used data-independent acquisition (DIA) proteomics to systematically analyze protein expression profiles in serum, follicular fluid (FF), and ovarian tissues of dairy cows. The goal was to identify potential molecular regulatory mechanisms and key biological pathways involved in IO. Serum, FF, and ovarian tissue samples were collected from six healthy and six IO multiparous Holstein cows at 63 (± 3) days postpartum. These samples were analyzed using DIA-based quantitative proteomics. In total, 155, 171, and 157 differentially expressed proteins (DEPs, P < 0.05 and FC ≥ 1.2 or ≤ 0.83) were identified in serum, FF, and ovarian tissues, respectively. Among these, 67 DEPs were upregulated and 88 were downregulated in serum, 38 were upregulated and 133 downregulated in FF, and 71 were upregulated and 86 downregulated in ovarian tissue. Integrated analysis revealed seven consistently altered proteins (APCS, KLKB1, HRG, TTR, CTSS, RBP4, and CUTA), of which CTSS was upregulated, while the others were downregulated across all sample types. Functional enrichment analysis indicated that most metabolic pathways, particularly glycolysis/gluconeogenesis, the tricarboxylic acid cycle, carbon metabolism, starch and sucrose metabolism, and amino acid biosynthesis and degradation (including branched-chain amino acid pathways), were markedly downregulated in IO cows, suggesting metabolic suppression. In contrast, cholesterol metabolism and complement and coagulation cascades showed a general upregulation trend. This study, for the first time, systematically characterizes coordinated alterations in key metabolic and reproductive pathways of IO cows through integrated multi-tissue proteomics. The findings provide insight into the molecular mechanisms of IO and suggest potential biomarkers and targets for improving ovarian function.

Ectopic fetus is defined as a pathological condition characterized by the presence of a fetus at an anatomically abnormal location within the maternal organism. This case series describes the clinical and pathological findings in three cats with ectopic fetuses. Diagnosis was based on radiographic findings and confirmed surgically and histologically. In one case, a Intratubal ectopic fetus with calcified fetuses was identified, which, to our knowledge, has not been previously documented in cats. In the other two cases, abdominal mummified fetuses were found attached to the greater omentum; one was an incidental finding during ovariohysterectomy, and the other was discovered six months post-ovariohysterectomy. Histopathological examination confirmed the pathological nature of the ectopic fetuses, and all cases were successfully treated via surgical resection of the ectopic tissue.

This paper reports a case of feline cholangiocarcinoma, a relatively rare condition in veterinary clinical practice. A 4-year-old female British Shorthair cat was presented with a 10-day history of anorexia and lethargy. An irregularly shaped mass was palpable on abdominal palpation. Diagnostic imaging and serum biochemical findings indicated extrahepatic biliary obstruction, and the diagnosis of cholangiocarcinoma was confirmed by histopathology and immunohistochemistry. Preoperative imaging in this cat indicated that the mass had not invaded major blood vessels such as the portal vein, and no evidence of distant metastasis was detected. The mass was surgically resected, followed by choledochoduodenostomy. The cat regained appetite on postoperative day 1, jaundice resolved within 48 hours, and biochemical parameters returned to normal by postoperative day 4. The cat was discharged 10 days after surgery, and no recurrence or metastasis was identified during the 3-month follow-up. This case documents the diagnostic and surgical management of feline extrahepatic cholangiocarcinoma and the short-term outcome following mass resection and biliary reconstruction.

Lycopene is a carotenoid with strong antioxidant activity, yet information on its use in waterfowl is limited. This study evaluated the effects of dietary lycopene on growth performance, antioxidant status, immune-related biomarkers, intestinal morphology, and cecal microbiota in growing geese. Seventy-two 35-d-old male Zi geese were allocated to two dietary treatments (6 pens/treatment, 6 birds/pen) and fed either a basal diet (CON) or the basal diet supplemented with 200 mg/kg lycopene (LYC) for 21 d under controlled temperature and humidity with ad libitum access to feed and water. Growth performance was analyzed by GLM with initial BW as a covariate (pen as the experimental unit), whereas antioxidant/immune indices were analyzed by two-way ANOVA with Tukey's test (bird as the experimental unit). Microbiota β-diversity was evaluated using Bray-Curtis distance with ordination (PCoA/NMDS) and permutation-based testing. Microbiota correlations were FDR-corrected. Differences were considered significant at P < 0.05. Compared with CON, LYC increased final body weight (2.77 vs. 2.29 kg, P < 0.05), average daily gain (66.67 vs. 43.80 g/d, P < 0.05), and reduced feed conversion ratio (4.38 vs. 6.22, P < 0.05). Lycopene improved antioxidant status, as indicated by higher serum CAT activity (day 21: 32.8 vs. 26.3, P < 0.01), T-AOC (day 21: 6.0 vs. 3.7, P < 0.01), and lower serum MDA (day 21: 3.9 vs. 6.1, P < 0.01); jejunal CAT was also higher at Day 21 (33.3 vs. 26.1, P < 0.05). Lycopene altered immune-related biomarkers, increasing serum IgA (day 21: 1.02 vs. 0.85, P < 0.05), IgM (day 21: 0.90 vs. 0.80, P < 0.05), and lysozyme activity (day 21: 20.3 vs. 18.3, P < 0.05), while decreasing IL-6 (day 21: 20.3 vs. 23.4, P < 0.05) and IFN-γ (day 21: 10.7 vs. 13.7, P < 0.05). Intestinal morphology was improved, with higher villus height-crypt depth ratios in the duodenum (day 21: 4.19 vs. 3.58, P < 0.05), jejunum (day 21: 4.34 vs. 3.99, P < 0.05) and ileum (day 21: 4.06 vs. 3.77, P < 0.05). Cecal microbiota differed between groups, with a higher relative abundance of Bacteroidota in LYC (47.96% vs. 41.35%, P < 0.05) and a positive correlation with serum T-AOC (P < 0.05). Functional prediction (PICRUSt2) suggested enrichment of nitrogen metabolism and amino sugar and nucleotide sugar metabolism pathways in LYC. Overall, dietary lycopene at 200 mg/kg improved growth performance and was associated with enhanced antioxidant capacity and shifts in immune-related biomarkers, intestinal morphology, and gut microbiota in geese.

Cisplatin (CIS) is a broad-spectrum anticancer drug widely used in the clinic; however, one of its side effects is that it can cause intestinal damage such as loss of appetite, vomiting, and diarrhea in patients. Epigallocatechin gallate (EGCG) is one of the main active substances in green tea, which has the effects of antitumor multiple drug resistance, antioxidation, and antiinflammatory properties. The aim of this study was to explore the protective effect of EGCG on CIS-induced intestinal injury in rats. First, physiological indices and HE staining indicated that compared with the control group, the physiological state of rats in the CIS group was worse, and the intestinal tissue was damaged, especially the ileum. In contrast, pretreatment with EGCG (20, 40, and 80 mg/kg) effectively alleviated the intestinal damage induced by CIS, with the 40 mg/kg dose demonstrating the most substantial protective effect. Additionally, 40 mg/kg EGCG pretreatment mitigated CIS-induced morphological and ultrastructural damage to intestinal tissues, reduced bacterial translocation, and preserved the integrity of the intestinal barrier. This treatment also altered the abundance of 19 bacterial species, including Lactobacillus and Shigella, and influenced amino acid metabolism and 15 metabolic pathways, including vitamin B6 metabolism by 16S RNA and metabolome sequencing. Furthermore, the expression of proteins associated with autophagy and the NRF2/Keap1 signaling pathway was inhibited. Lastly, ML385 (NRF2 signaling pathway inhibitor) reversed the protective effects of EGCG. Taken together, our findings indicate that EGCG ameliorates CIS induced hepatoenteric toxicity in rats by regulating the intestinal flora and targeting the Nrf2/Keap1 signal axis.

The effects of dietary supplementation with epigallocatechin-3-gallate (EGCG) on growth performance, antioxidant capacity, immune function, and gut microbiota in geese were investigated. Seventy-two healthy 35-day-old male geese were randomly divided into a control group (basal diet) or an EGCG group (basal diet + 200 mg/kg EGCG), with 36 geese per group, which was further subdivided into 6 replicates (6 geese per replicate). The experiment lasted 21 days. Geese in the EGCG group exhibited significantly higher final body weight (2.93 kg vs. 2.28 kg, P < 0.01) and average daily gain (72.38 g/d vs. 41.4 g/d, P < 0.01) along with a 42.8 % reduction in the feed conversion ratio (3.95 vs. 6.91, P < 0.01) versus the control. Liver weights in the EGCG group were significantly elevated compared to the CON group on days 14 and 21 (P < 0.05) and a strong correlation between liver weight and body weight. EGCG significantly increased catalase, glutathione peroxidase, and superoxide dismutase activities, and the total antioxidant capacity in serum and jejunum while decreasing malondialdehyde (MDA) levels (P < 0.05). Immunological analyses revealed elevated serum immunoglobulin (Ig)A, IgG, and IgM and lysozyme in the EGCG group (P < 0.05), accompanied by a decrease in the pro-inflammatory cytokines interleukin-6 and interferon-γ. Intestinal morphology demonstrated increased villus height–to–crypt depth ratios in the duodenum and ileum (P < 0.05). 16S rRNA sequencing indicated that EGCG increased the relative abundance of Akkermansia and Verrucomicrobiota (P < 0.05) in the cecal content and enriched microbial functions related to inorganic ion transport and metabolism. Correlation analysis revealed positive associations between Akkermansia and IgA, and between Firmicutes and oxidative damage markers (MDA). Overall, dietary supplementation with 200 mg/kg EGCG could improve growth performance, antioxidant capacity, immune response, and gut microbiota structure in geese, supporting its potential as a plant-based feed additive.

Foot rot is a contagious disease caused by F.necrophorum. It is responsible for economic losses in dairy farming. Studies on foot rot in dairy cows are focused on the isolation and identification of pathogens and treatment methods. Few studies have reported inflammatory changes in tissues and regulatory mechanisms following infection. Here, the effects of F.necrophorum infection on the skin explants and skin fibroblasts between the toes of cattle were analyzed using histopathology and other techniques. F.necrophorum infection increased the epidermal thickness and number of hair follicles and sebaceous glands. Other skin appendages exhibited varying degrees of necrosis, and a significant infiltration of inflammatory cells was noted in the interdigital skin explants. The expressions of pro-inflammatory cytokines (IL-1β and TNF-α) and key genes in the inflammatory signalling pathway (TNFR1 and NF-κB p65) were elevated. Treatment with the TNFR1 inhibitor CAY10500 reduced inflammatory cell infiltration and alleviated TNFR1 and p65 expression. An inflammatory cell model was established using different proportions of F.necrophorum to infect BDF cells. F.necrophorum infection significantly inhibited the proliferation and viability of BDF cells and enhanced the expression of TRADD, TRAF2, TNF-α, and IL-18. CAY10500 reduced the F.necrophorum infection-induced inflammatory response and induced inflammatory responses in interdigital skin explants and BDF cells by inhibiting the TNF-α/TNFR1/NF-κB signaling pathway. In summary, these findings provide new insights into the mechanism of inflammatory responses in dairy cows with foot rot.

Acute pancreatitis (AP) is a common exocrine pancreatic disease that can lead to systemic inflammatory response syndrome and multiorgan failure in canines. The therapeutic benefits of adipose-derived stem cells (ADSCs) and conditioned medium (CM) and the role in ferroptosis regulation in managing AP in canines (dogs) were investigated in this study. Sixteen dogs were randomly divided into a control (CON), AP, ADSC, or ADSC-CM group. The AP model was established by injecting the dogs with sodium taurocholate (5

Osteoarthritis is often exacerbated by obesity and metabolic syndrome, which are frequently associated with high-fat diets (HFD). This study investigates the direct effects of HFD on articular cartilage following anterior cruciate ligament transection (ACLT) in rats, while also examining the role of free fatty acids (FFA) on chondrocyte activity. Our findings indicate that short-term HFD treatment after ACLT reduces pain sensitivity, alleviates knee swelling, and mitigates cartilage damage. Joint imaging studies, along with serum analyses of inflammatory markers and extracellular matrix (ECM) degradation, further underscore the protective role of post-surgical HFD in maintaining cartilage integrity. Notably, HFD significantly enhances AMP-activated protein kinase (AMPK) phosphorylation in cartilage. In vitro experiments reveal that low concentrations of FFAs stimulate chondrocyte proliferation and energy metabolism, whereas AMPK inhibition leads to elevated expression of inflammatory mediators and ECM-degrading enzymes in chondrocytes. Collectively, these results suggest that short-term HFD following ACLT surgery exerts protective effects on cartilage, primarily through AMPK activation.

This article describes a Rough-legged Buzzard (Buteo lagopus) from the Wildlife Nature Reserve in Daqing, China. According to the staff of Daqing Wildlife Nature Reserve, the Rough-legged Buzzard was found to have impaired flight ability, depression, and loss of aggression, but its feathers were full and there were no obvious external injuries, so it was sent to the Wildlife Rehabilitation Center of the Teaching Animal Hospital of Heilongjiang Bayi Agricultural University for treatment. After the vet conducted X-ray and blood tests, he diagnosed the bird with a foreign body in the crop and crop inflammation. He finally treated the bird using endoscopic techniques, and the bird recovered well and was successfully released.

Cisplatin (CDDP, CIS) is the first generation of platinum-containing drugs. However, the metabolites of CDDP and other chemotherapeutic drugs cause serious damage to intestinal mucosa and damage intestinal mucosal barrier, leading to dose-limiting side effects such as diarrhea. China is the home of tea, among the many types of tea, green tea is the most widely studied, its anti-cancer, anti-obesity and neuroprotective effects. First, rats were given EGCG(40 mg/kg) 28 d and then injected CDDP ༈7 mg/kg༉ to explore the protective effect of EGCG. HE staining, immunohistochemistry, transmission electron microscopy and TUNEL staining showed that pretreatment of EGCG could reduce intestinal pathological injury, intestinal dysfunction, oxidative stress, apoptosis and hyperautophagy induced by CDDP in rats. Then, it was confirmed by immunofluorescence and Western blot tests that EGCG protected the intestine mainly by activating the NRF2/HO-1 signaling pathway. Finally, the protective effect of EGCG was weakened after administration of NRF2 inhibitor (ML385) for 28 d and then injection of cisplatin. In summary, EGCG can reduce intestinal oxidative damage, apoptosis and autophagy through Nrf2/HO-1 signaling pathway, thus playing a role in intestinal protection, providing a new idea for the prevention of side effects of chemotherapy drugs.

Osteoarthritis (OA) is the most common joint disease, causing symptoms such as joint pain, swelling, and deformity, which severely affect patients’ quality of life. Despite advances in medical treatment, OA management remains challenging, necessitating the development of safe and effective drugs. Quercetin (QUE), a natural flavonoid widely found in fruits and vegetables, shows promise due to its broad range of pharmacological effects, particularly in various degenerative diseases. However, its role in preventing OA progression and its underlying mechanisms remain unclear. In this study, we demonstrated that QUE has a protective effect against OA development both in vivo and in vitro, and we elucidated the underlying molecular mechanisms. In vitro, QUE inhibited the expression of IL−1β-induced chondrocyte matrix metalloproteinases (MMP3 and MMP13) and inflammatory mediators such as INOS and COX−2. It also promoted the expression of collagen II, thereby preventing the extracellular matrix (ECM). Mechanistically, QUE exerts its protective effect on chondrocytes by activating the SIRT1/Nrf−2/HO−1 and inhibiting chondrocyte ferroptosis. Similarly, in an OA rat model induced by anterior cruciate ligament transection (ACLT), QUE treatment improved articular cartilage damage, reduced joint pain, and normalized abnormal subchondral bone remodeling. QUE also reduced serum IL−1β, TNF−α, MMP3, CTX−II, and COMP, thereby slowing the progression of OA. QUE exerts chondroprotective effects by inhibiting chondrocyte oxidative damage and ferroptosis through the SIRT1/Nrf−2/HO−1 pathway, effectively alleviating OA progression in rats.

In order to explore the therapeutic effect of Resveratrol (Res)-activated Natural Killer (NK) cells on canine mammary tumors, this study employed a range of assays, including wound healing, colony formation, Transwell, flow cytometry, and Western blot experiments, to investigate the impact of Res-pretreated NK cells on canine mammary tumor cells in vitro. Additionally, a tumor-bearing mouse model was utilized to further analyze the therapeutic effects of Res-pretreated NK cells in vivo. The results showed that Res enhances the capacity of NK cells to induce apoptosis, pyroptosis, and ferroptosis in canine breast tumor cells, while also augmenting their influence on the migration, invasion, and epithelial-mesenchymal transition of these cells. Furthermore, pretreatment of NK cells with Res significantly amplified their inhibitory effect on breast tumor growth in vivo and promoted tumor tissue apoptosis. Additionally, Res enhanced the recruitment of NK cells to other immune cells in the body. In summary, Res has been shown to enhance the anti-breast-tumor effect of NK cells both in vitro and in vivo, offering a new avenue for optimizing immunotherapy for canine breast tumors.

In order to explore the metabolic and ionic changes of hoof-deformed cows, the serum samples of 10 healthy cows (group C) and 10 hoof-deformed cows (group T) were analyzed by LC-MS/MS and ICP-OES/MS. The pathway enrichment of differential metabolites was analyzed by screening and identifying differential metabolites and ions and using a bioinformatics method. The integration of metabolomics and ionics was analyzed with ggplot2 software in R language, and verified by MRM target metabolomics. The results showed that 127 metabolites were screened by metabolomics, of which 81 were up-regulated (p < 0.05) and 46 were down-regulated (p < 0.05). The results of ICP-OES/MS showed that 13 kinds of ions such as K, Li, and Pb in serum of dairy cows were up-regulated, while 18 kinds of ions such as Al, Cu and Sb were down-regulated. The integrated analysis of metabolomics and ionics found that potassium ions were positively correlated with L-tyrosine, L-proline, thiamine and L-valine. Sodium ions were positively correlated with L-valine and negatively correlated with α-D-glucose. The results of high-throughput target metabolomics showed that the contents of L-proline, L-phenylalanine and L-tryptophan in serum of dairy cows increased significantly, which was consistent with the results of non-target metabolomics. In a word, the metabolism and ion changes in dairy cows with hoof deformation were revealed by metabolomics and ionics.

临床思维能力是临床操作能力的核心和基础,是成为一名专业的兽医诊疗人才的必备能力,临床思维的好坏直接影响动物疾病诊疗结果的准确化和专业化.结合动物医学专业兽医病理学课程的教学实际情况,以临床思维能力的培养和提高为目标,在兽医病理生理学、兽医病理解剖学、机能学实验和形态学实验等四门课程教学过程中,通过提倡PBL教学模式和传统教学模式的结合、开展兽医病理分析讨论会、建立及使用教学特色资源库、利用虚拟仿真教学平台、构建"虚实结合"实验教学新模式和"三全"教学考核评价体系等一系列教学改革措施,构建兽医病理学教学新体系并进行推广实践.该教学新体系的实施激发了学生的学习兴趣,提高了教学质量,为培养具有扎实理论知识、良好职业素养、具备较强临床操作能力和临床思维能力的动物疾病诊疗技术人员奠定基础.

Canine distemper (CD), caused by canine distemper virus (CDV), is a highly contagious and lethal disease in domestic and wild carnivores. Although CDV live-attenuated vaccines have reduced the incidence of CD worldwide, low levels of protection are achieved in the presence of maternal antibodies in juvenile animals. Moreover, live-attenuated CDV vaccines may retain residual virulence in highly susceptible species and cause disease. Here, we generated several CDV DNA vaccine candidates based on the biscistronic vector (pIRES) co-expressing virus wild-type or codon-optimized hemagglutinin (H) and nucleocapsid (N) or ferret interferon (IFN)-γ, as a molecular adjuvant, respectively. Apparently, ferret (Mustela putorius furo)-specific codon optimization increased the expression of CDV H and N proteins. A ferret model of CDV was used to evaluate the protective immune response of the DNA vaccines. The results of the vaccinated ferrets showed that the DNA vaccine co-expressing the genes of codon-optimized H and ferret IFN-γ (poptiH-IRES-IFN) elicited the highest anti-CDV serum-neutralizing antibodies titer (1:14) and cytokine responses (upregulated TNF-α, IL-4, IL-2, and IFN-γ expression) after the third immunization. Following vaccination, the animals were challenged with a lethal CDV 5804Pe/H strain with a dose of 105.0 TCID50. Protective immune responses induced by the DNA vaccine alleviated clinical symptoms and pathological changes in CDV-infected ferrets. However, it cannot completely prevent virus replication and viremia in vivo as well as virus shedding due to the limited neutralizing antibody level, which eventually contributed to a survival rate of 75% (3/4) against CDV infection. Therefore, the improved strategies for the present DNA vaccines should be taken into consideration to develop more protective immunity, which includes increasing antigen expression or alternative delivery routes, such as gene gun injection.

疝是内脏或组织从自然孔道或病理性破裂孔至皮下或其他解剖腔的疾病.膈疝是腹腔内一种或几种内脏器官通过横膈的破裂孔进入胸腔的一种病理状态,多由膈的腱质部或肌质部受到意外损伤或膈先天性缺损所致.当发生膈疝时应尽快治疗,否则将因呼吸窘迫而危及生命.1只3岁成年猫连续多日呕吐、腹泻、食欲减退、呼吸困难,采用实验室检查和影像学检查,结合患猫病史和临床症状,确诊该猫患有膈疝,疝内容物为胆囊.通过外科手术进行器官复位及膈疝修复,手术的治疗效果理想.

This study sought to establish a real-time reverse transcription (RT)-PCR method to differentially detect canine distemper virus (CDV) wild-type and vaccine strains. To this end, a pair of CDV universal primers and two specific minor groove binder (MGB) probes, harboring a T/C substitution in the hemagglutinin (H) gene, were designed. Using a recombinant plasmid expressing the H gene of the CDV wild-type or vaccine strain as standards, a sensitive and specific multiplex real-time RT-PCR was established for quantitative and differential detection of CDV wild-type and vaccine strains. The limit of detection for this multiplex assay was 22.5 copies/μL and 2.98 copies/μL of viral RNA for wild-type and vaccine strains, respectively. Importantly, the wild-type and vaccine MGB probes specifically hybridized different genotypes of wild-type CDV circulating in China as well as globally administered vaccine viruses, respectively, with no cross-reactivity observed with non-CDV viruses. Moreover, this method was successfully applied for the quantitative detection of CDV RNA in tissue samples of experimentally infected breeding foxes, raccoon dogs, and minks. Additionally, the multiplex real-time RT-PCR was able to detect the viral RNA in the whole blood samples as early as 3 days post-infection, 3 to 4 days prior to the onset of clinical signs in these CDV infection animals. Hence, the established multiplex real-time RT-PCR method is useful for differentiating wild-type CDV and vaccine strains in China, and for conducting canine distemper early diagnosis as well as dynamic mechanism of CDV replication studies in vivo.