Veterinary microbiology最新文献_第10页

Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants 用于山羊布鲁氏菌病血清诊断的室内iELISA的验证和用于检测反刍动物抗光滑布鲁氏菌特异性抗体的瘤b菌裂解物的性能评价。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-15 DOI: 10.1016/j.vetmic.2025.110389

Camila N. Foster , Ursula A. Rossi , Carlos A. Rossetti

Enzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-Brucella antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using B. melitensis whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 B. neotomae in replacement of BSL-3 Brucella species as an antigen for the detection of Brucella-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (k = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 Brucella for antigen preparation, we replaced it with a less virulent Brucella species such as B. neotomae. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the B. neotomae whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.

酶联免疫吸附试验（ELISA）是一种广泛应用的检测血清中抗布鲁氏菌抗体的有效方法，操作简便，灵敏度和特异性高。在这项研究中，我们验证了一种用羊分枝杆菌全细胞裂解液作为抗原（Bm-WCL iELISA）的内部间接ELISA用于山羊布鲁氏菌病的血清诊断，并评估了用BSL-2新瘤分枝杆菌代替BSL-3布鲁氏菌作为抗原检测反刍动物血清中布鲁氏菌特异性抗体的效果。利用724份经缓冲板抗原（BPA）和补体固定（CF）试验均为布鲁氏菌病阳性或阴性的杂交山羊血清样本，成功验证了bmwcl - iELISA的敏感性（Se）为91.83 %(88.51 ~ 94.25 %)，特异性（Sp）为97.41 %（95.41 ~ 98.70 %）。此外，Bm-WCL iELISA在217份血清样本中与商业iELISA试剂盒（k = 0.94）显示出很大的一致性。为了避免使用BSL-3布鲁氏菌进行抗原制备，我们将其替换为毒性较低的布鲁氏菌，如新瘤芽胞杆菌。采用瘤芽胞杆菌全细胞匀浆（Bn-WCL） elisa对214份山羊和220份牛血清进行布鲁氏菌病诊断。ROC曲线分析表明，山羊和牛血清Se和Sp的临界值分别为63.83 PP和24.04 PP，山羊血清Se和Sp的相关值分别为98.18 %（93.61 ~ 99.68 %）和90.38 %(83.20 ~ 94.69 %)，牛血清Se和Sp的相关值分别为95.45 %（89.80 ~ 98.04 %）和96.36 %（91.02 ~ 98.58 %）。这些结果证实了内部Bm-WCL iELISA的效用，并鼓励在资源有限的布鲁氏菌病流行地区对Bn-WCL iELISA进行血清诊断。

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引用次数: 0

Sequential emergence and quantitative dynamics of key bacterial species preceding digital dermatitis lesion onset in dairy cattle

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-14 DOI: 10.1016/j.vetmic.2025.110378

Angelica Petersen Dias, Karin Orsel, Corienne Sarah Gammariello, Jeroen De Buck

Digital dermatitis (DD) is a skin infection of cattle’s feet with multiple bacteria suspected to be involved, yet its precise etiopathogenesis remains unclear. This longitudinal study explored the temporal changes of seven DD-associated bacteria in feet developing lesions or remaining healthy, while simultaneously investigating their persistence in potential reservoirs as sources of infection. Weekly swabs were collected from feet skin and saliva of 53 Holstein cows without DD lesions sequentially enrolled at calving in a commercial dairy herd. At the end of the study, samples from all cases and a subset of matched controls were analyzed (1:2 ratio) at five-time points (weeks −3, −2, −1, 0 - when early signs of DD were observed - and +1) and subjected to qPCR targeting Treponema phagedenis, T. medium, T. pedis, Porphyromonas levii, Bacteroides pyogenes, Fusobacterium necrophorum, and F. mortiferum. Linear mixed-effect models assessed the bacterial number changes within cows (cases) and between cows (cases vs controls). Throughout the study, 8 cows developed signs of DD. P. levii, F. necrophorum, and B. pyogenes numbers increased two weeks before the first visible lesion. T. phagedenis and T. pedis numbers increased one week before, suggesting a sequential colonization and potential synergism in triggering DD. Only P. levii and F. necrophorum were persistently present in saliva and skin, while Treponema spp. persisted solely in lesions. Our results inform specific bacterial dynamics associated with DD pathogenesis and might advise future attempts to effectively treat and control DD.

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引用次数: 0

Exosomal miR-27a-5p attenuates inflammation through Toll-like receptor 7 in foodborne Salmonella infections 在食源性沙门氏菌感染中，外泌体miR-27a-5p通过toll样受体7减轻炎症。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-14 DOI: 10.1016/j.vetmic.2025.110394

Mingjuan Qu , Shengfa Su , Linlin Jiang , Xin Yu , Jianlong Zhang , Hongwei Zhu , Kexue Han , Xingxiao Zhang

Salmonella is a common food-borne pathogen that is highly pathogenic and infectious, causing serious harm to livestock breeding and food safety. Uncovering the mechanisms of Salmonella infection and immune evasion can effectively prevent Salmonella contamination of livestock and poultry food. Here, small RNA sequencing results showed that exosomes produced by naïve murine macrophages RAW 264.7 cells contained a unique enrichment of a set of microRNAs (miRNAs) after Salmonella infection. Quantitative real-time polymerase chain reaction (qPCR) analysis verified that the tested miRNA (i.e. miR-27a-5p, miR-92a-1-5p and miR-1249-5p) showed similar expression patterns, consistent with small RNA sequencing data. TargetScan database predicted that the most promising targets for the differentially expressed miRNAs were abundant in the immune system, infectious diseases, and signal transduction pathways. Dual-luciferase reporter assays confirmed that Toll-like receptor 7 (TLR7) was the target of miR-27a-5p. Western blotting and enzyme-linked immunosorbent assay (ELISA) results revealed that overexpression of miR-27a-5p suppressed inflammation by targeting TLR7/nuclear factor kappa-B (NF-κB) signaling pathway and leading interleukin-6 (IL-6) and IL-1β cytokines slightly reduction in recipient macrophages, suggesting that exosomal miR-27a-5p uptake by naïve macrophages may inhibit pro-inflammatory macrophage differentiation. Therefore, these results contribute to our systematic understanding of the mechanism of exosomal miRNA in Salmonella infection, providing a potential target for preventing immune escape from Salmonella.

沙门氏菌是一种常见的食源性致病菌，具有高致病性和传染性，对家畜养殖和食品安全造成严重危害。揭示沙门氏菌感染和免疫逃逸的机制，可以有效预防沙门氏菌污染畜禽食品。这里，小RNA测序结果显示naïve小鼠巨噬细胞RAW 264.7细胞产生的外泌体在沙门氏菌感染后含有一组独特的microRNAs （miRNAs）富集。定量实时聚合酶链反应（qPCR）分析证实，检测的miRNA（即miR-27a-5p， miR-92a-1-5p和miR-1249-5p）表现出相似的表达模式，与小RNA测序数据一致。TargetScan数据库预测，这些差异表达的mirna最有希望的靶标在免疫系统、传染病和信号转导途径中丰富。双荧光素酶报告基因检测证实toll样受体7 （TLR7）是miR-27a-5p的靶标。Western blotting和酶联免疫吸附试验（ELISA）结果显示，过表达miR-27a-5p通过靶向TLR7/核因子κ b （NF-κB）信号通路和导致受体巨噬细胞中白细胞介素-6 （IL-6）和IL-1β细胞因子的轻微降低来抑制炎症，提示naïve巨噬细胞摄取外泌体miR-27a-5p可能抑制促炎巨噬细胞的分化。因此，这些结果有助于我们系统地了解外泌体miRNA在沙门氏菌感染中的作用机制，为防止沙门氏菌的免疫逃逸提供潜在的靶点。

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引用次数: 0

Glaesserella parasuis serotype 5 promotes pyroptosis via degrading Caveolin-1 in 3D4/21 cells 血清型5副猪小绿杆菌通过降解Caveolin-1促进3D4/21细胞的焦亡。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-13 DOI: 10.1016/j.vetmic.2025.110393

Huixing Lin , Jianan Zhang , Qing Wang , Hong Zhou , Hongjie Fan

Glaesserella parasuis (G. parasuis) is an important pathogen, which can cause systemic inflammatory response in pigs and bring huge economic losses to the global swine industry. G. parasuis can induce a strong inflammatory response in the lungs under environmental changes and certain stress conditions. However, the underlying mechanism of this adverse response has not been thoroughly studied. In this study we demonstrated that G. parasuis serotype 5 strain (GPS5-SQ) has the potential to induce pyroptosis in 3D4/21 cells. GPS5-SQ could degrade the expression of Cav-1. Knockdown or overexpression of Cav-1 promoted or reduced the occurrence of pyroptosis, respectively. These results suggested that Cav-1 is involved in pyroptosis induced by GPS5-SQ in 3D4/21 cells. In addition, overexpression of Cav-1 suppressed the activation of NLRP3 inflammasome by inhibiting ASC oligomerization, resulted in reducing pyroptosis. In general, we found that GPS5-SQ infection could promote pyroptosis by degrading the expression of Cav-1. The results of the study revealed the new mechanism of inflammation induced by GPS5-SQ in 3D4/21 cells.

副猪Glaesserella parasuis （G. parasuis）是一种重要的病原体，可引起猪的全身炎症反应，给全球养猪业带来巨大的经济损失。副猪绦虫在环境变化和一定应激条件下可引起肺部强烈的炎症反应。然而，这种不良反应的潜在机制尚未得到充分研究。在本研究中，我们证实了副猪螺旋体血清5型菌株（GPS5-SQ）在3D4/21细胞中具有诱导焦亡的潜力。GPS5-SQ可降低Cav-1的表达。敲低或过表达Cav-1分别促进或减少焦亡的发生。提示Cav-1参与了GPS5-SQ诱导3D4/21细胞的焦亡。此外，过表达Cav-1通过抑制ASC寡聚化抑制NLRP3炎性体的激活，导致焦亡减少。总的来说，我们发现GPS5-SQ感染可以通过降低Cav-1的表达来促进焦亡。本研究结果揭示了GPS5-SQ诱导3D4/21细胞炎症的新机制。

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引用次数: 0

Sus scrofa RNase L inhibits PRRSV replication by activation of type I IFN signaling pathway and apoptosis Sus scrofa RNase L通过激活I型IFN信号通路和细胞凋亡抑制PRRSV复制。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-13 DOI: 10.1016/j.vetmic.2025.110392

Xiangju Wu , Xiaoyan Cong , Ping Jiang , Mingming Zhou , Ying Yu , Dandan Jiang , Yue Hu , Juntong Li , Jinxia Zhang , Ying Cao , Haowen Zhang , Yijun Du , Jing Qi

Porcine reproductive and respiratory syndrome virus (PRRSV) has become one of the most economically important diseases to the global pig industry. RNase L is a ubiquitous cellular endoribonuclease that is activated upon the binding of a specific ligand, 2′,5′-linked oligoadenylates (2–5 A), which is synthesized by oligoadenylate synthetases (OASs). However, whether Sus scrofa RNase L (sRNase L) could inhibit PRRSV replication and its mechanism have not been fully elucidated. In this study, sRNase L was cloned and characterized in homology and structure firstly. Then the antiviral activity of sRNase L against PRRSV was explored. Overexpression of sRNase L significantly inhibited the propagation of PRRSV when treated with 2–5 A or poly(I: C) or mock treated. Furthermore, sRNase L induced degradation of cellular and viral ssRNAs, enhanced the activation of IFN-β promoter and IFN-β expression, and induced apoptosis to inhibit PRRSV replication. Taken together, we have first elucidated the anti-PRRSV function and the underlying mechanism of sRNase L, which may provide a new strategy for preventing PRRSV infection.

猪繁殖与呼吸综合征病毒（PRRSV）已成为全球养猪业最重要的经济疾病之一。RNase L是一种普遍存在的细胞核糖核酸内切酶，在与特定配体结合时被激活，2',5'-连接的低聚腺苷酸（2-5 a），由低聚腺苷酸合成酶（OASs）合成。然而，sRNase L是否能够抑制PRRSV的复制及其机制尚未完全阐明。本研究首次克隆了sRNase L，并对其同源性和结构进行了鉴定。探讨sRNase L对PRRSV的抗病毒活性。sRNase L的过表达在2-5 A或poly（I: C）或mock处理下显著抑制PRRSV的繁殖。此外，sRNase L诱导细胞和病毒的ssrna降解，增强IFN-β启动子的激活和IFN-β的表达，诱导细胞凋亡，从而抑制PRRSV的复制。综上所述，我们首次阐明了sRNase L的抗PRRSV功能及其潜在机制，这可能为预防PRRSV感染提供新的策略。

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引用次数: 0

Concurrent but consecutive vaccination of modified live PRRSV-1 and PRRSV-2 provides better protection in nursery pigs 同时但连续接种改良的 PRRSV-1 和 PRRSV-2 活疫苗可为保育猪提供更好的保护。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-13 DOI: 10.1016/j.vetmic.2025.110391

Yashavanth Shaan Lakshmanappa , Pengcheng Shang , Sankar Renu , Santosh Dhakal , Bradley Hogshead , Yihong Xiao , Tao Wang , Ying Fang , Gourapura J. Renukaradhya

Porcine reproductive and respiratory syndrome (PRRS) virus is a severe threat to the global swine industry. Modified live virus vaccines (MLVs) for two PRRSV species (PRRSV-1 and PRRSV-2) are the most widely used approach to control PRRSV-caused diseases. For swine herds influenced by PRRSV-1 and PRRSV-2, how to rationalize MLV immunization strategies for robust and cross-protective immune responses has been a long-lasting need. In this study, we found that the replication of PRRSV-1 is strongly suppressed by co-infection with PRRSV-2 in vitro, especially under concurrent co-infection conditions. We compared the adaptive immune responses between consecutive and concurrent vaccination methods in nursery pigs, vaccinated either 3 days apart (PRRSV-1 MLV followed by PRRSV-2 MLV, consecutive) or together on the same day (concurrent). PRRSV-1 RNAs were mainly detectable in the sera of consecutively vaccinated pigs. In contrast, PRRSV-2 RNAs in sera were not changed in both vaccination strategies. After the homologous PRRSV-1 or PRRSV-2 challenge, we found that consecutive vaccination slightly improved PRRSV-1 viremia clearance and did not attenuate the PRRSV-2 viremia clearance. Both vaccination strategies induced comparable T-helper cell responses against PRRSV-1 and PRRSV-2 in peripheral blood before and after the challenge. Interestingly, consecutive vaccination induced significantly higher PRRSV-1-specific post-challenge T-helper and cytotoxic T cells responses in the tracheobronchial lymph nodes than concurrent vaccination. Furthermore, consecutive vaccination significantly improved neutralizing antibody responses against PRRSV-1 and PRRSV-2 in comparison with concurrent vaccination. In conclusion, consecutive vaccination appears to be better for viral clearance and induction of adaptive immune response, and our study provides a preliminary rationale to optimize PRRS MLV immunization strategy for better dual protection.

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引用次数: 0

Effectiveness of a single-dose phage cocktail on the reduction of multidrug-resistant Escherichia coli in suckling piglets 单剂量噬菌体鸡尾酒对减少哺乳仔猪多重耐药大肠杆菌的效果。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-13 DOI: 10.1016/j.vetmic.2025.110395

Viphavanh Chanthavong , Nattha Vigad , Wattana Pelyuntha , David Yembilla Yamik , Kitiya Vongkamjan , Mingkwan Yingkajorn , Warangkhana Chaisowwong , Kittiphong Tippaya , Phacharaporn Tadee , Kridda Chukiatsiri

Antibiotics are commonly used in pig farming to control infections caused by diarrhea-causing Escherichia coli (E. coli). However, improper or excessive use of antibiotics in pigs can enhance antibiotic resistance (ABR). This study used bacteriophage (phage) treatment to control ABR E. coli in diarrheal suckling piglets. Fifty E. coli isolates were previously isolated from suckling pigs, which showed resistance to amoxicillin (100 %), oxytetracycline and neomycin (94 %), sulfamethoxazole-trimethoprim (70 %), gentamicin (56 %), cephalexin (54 %), enrofloxacin (42 %), and colistin (28 %). Five phages (WPEC1, WPEC2, WPEC3, WPEC4, and WPEC5) were included in this study. These phages showed a diverse lytic profile ranging from 46.0 % to 64.0 % on the tested ABR E. coli isolates. The phage cocktail reduced the count of five representative E. coli by showing up to 8 log-units reduction (p < 0.05) after phage treatment for 6–24 h. From the in vivo study, a single dose of the phage cocktail (9 log PFU/mL) reduced the number of E. coli present in the gastrointestinal tract of suckling piglets by showing a 1.33 log-units reduction on day 7 (p < 0.05). In addition, the fecal score of the phage treatment group was lower than that of the control group (p < 0.05). However, body weight gain (BWG) and average daily gain (ADG) were not significantly different in both groups (p > 0.05). These findings suggest that a developed phage cocktail could be used as a potential biocontrol to fight ABR E. coli, reduce the chance of piglet mortality, and increase safety during pig production.

抗生素通常用于养猪业，以控制由引起腹泻的大肠杆菌引起的感染。然而，不适当或过量使用抗生素会增强猪的抗生素耐药性。本研究采用噬菌体（phage）处理控制腹泻哺乳仔猪ABR大肠杆菌。先前从乳猪中分离出50株大肠杆菌，对阿莫西林（100 %）、土霉素和新霉素（94 %）、磺胺甲氧唑-甲氧苄啶（70 %）、庆大霉素（56 %）、头孢氨苄（54 %）、恩诺沙星（42 %）和粘菌素（28 %）耐药。本研究共纳入5个噬菌体（WPEC1、WPEC2、WPEC3、WPEC4、WPEC5）。这些噬菌体在ABR大肠杆菌分离株上显示出不同的裂解谱，范围从46.0 %到64.0 %。噬菌体鸡尾酒减少了5个代表性大肠杆菌的计数，减少了8个对数单位（p  0.05）。这些发现表明，开发出的噬菌体鸡尾酒可作为一种潜在的生物防治手段来对抗ABR大肠杆菌，降低仔猪死亡率，提高生猪生产过程中的安全性。

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引用次数: 0

Occurrence and molecular identification of haemotropic Mycoplasma species in grey wolves (Canis lupus) from southern Europe 南欧灰狼（Canis lupus）嗜血支原体的发生及分子鉴定。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-12 DOI: 10.1016/j.vetmic.2025.110390

Susana Remesar , David Cano-Terriza , Patrocinio Morrondo , Álvaro Oleaga , Barbara Moroni , Nuno Santos , Serena Robetto , Lisa Guardone , Pablo Díaz , Saúl Jiménez-Ruiz , Joana Ferreira-e-Silva , Moisés Gonzálvez , Ignacio García-Bocanegra

Although wild and domestic carnivores share some haemotropic Mycoplasma species, information about the circulation of this pathogen in grey wolves (Canis lupus) populations is still very limited. Thus, a geographically broad-based investigation was performed for determining the occurrence and diversity of Mycoplasma spp. in three different wolf populations from southern Europe. Between 2001 and 2023, spleen samples from 285 grey wolves from Spain (n = 129), Italy (n = 113), and Portugal (n = 43) were collected. The presence of haemotropic Mycoplasma was assessed targeting the 16S rRNA gene using two PCR assays in parallel; in addition, the 16S–23S rRNA intergenic spacer was analysed for further identification of the positive samples. The influence of the sampling country, sex, and age of the animals on the prevalence of Mycoplasma spp. was also assessed by a generalized linear model analysis. The percentage of positive wolves was 13.3 % (38/285), and the occurrence was significantly higher in Spain (20.9 %) than in Italy (8.0 %) and Portugal (4.7 %). Mycoplasma haemocanis (10.5 %) and Candidatus M. haematoparvum (2.1 %), were identified; in addition, an uncultured Mycoplasma sp. was also detected (0.7 %). Our results confirm the circulation of potentially zoonotic Mycoplasma in wolf populations from southern Europe. To our knowledge, this is the first report of Ca. M. haematoparvum in wolves from Italy and Portugal. In addition, a Mycoplasma sp., previously found in dogs, has been detected for the first time in wolves. Further studies are needed to fully molecularly characterise haemotropic Mycoplasma spp., which will serve as a basis for the study of its ecoepidemiology.

尽管野生和家养食肉动物都有一些嗜血性支原体，但关于这种病原体在灰狼（Canis lupus）种群中传播的信息仍然非常有限。因此，进行了一项地理基础广泛的调查，以确定支原体在南欧三个不同狼种群中的发生和多样性。在2001年至2023年间，从西班牙（n = 129）、意大利（n = 113）和葡萄牙（n = 43）的285只灰狼身上采集脾脏样本。以16S rRNA基因为靶点，采用两种PCR方法检测嗜血性支原体的存在；此外，分析16S-23S rRNA基因间间隔，进一步鉴定阳性样品。通过广义线性模型分析，还评估了采样国家、性别和动物年龄对支原体流行率的影响。阳性率为13.3 %(38/285)，其中西班牙（20.9 %）明显高于意大利（8.0 %）和葡萄牙（4.7 %）。鉴定出血支原体（10.5% %）和血疟原虫（2.1% %）；此外，还检出未培养的支原体（0.7 %）。我们的结果证实了潜在的人畜共患支原体在南欧狼种群中的传播。据我们所知，这是首次在意大利和葡萄牙的狼中发现血疟原虫。此外，以前在狗身上发现的支原体sp.首次在狼身上发现。需要进一步的研究来充分表征嗜血性支原体的分子特征，这将为其生态流行病学研究奠定基础。

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引用次数: 0

Detection of equine influenza virus gene in the air around infected horses 感染马匹周围空气中马流感病毒基因的检测。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-11 DOI: 10.1016/j.vetmic.2025.110388

Manabu Nemoto , Nanako Kawanishi, Yoshinori Kambayashi, Hiroshi Bannai, Takashi Yamanaka, Koji Tsujimura

Equine influenza virus (EIV) can be transmitted by inhalation of aerosolized droplets, direct contact, and contaminated fomites. However, to our knowledge, there are no reports of the recovery of EIV from the air surrounding infected horses. Here, we evaluated whether EIV can be recovered from the air in the stalls of experimentally infected horses by using an air sampler. Furthermore, we examined whether rapid molecular test kits with reaction times of less than 30 min can detect EIV from air samples for potential field application. Two horses kept in individual stalls were experimentally infected with EIV. Air samples were collected daily by using an air sampler until 13 days post-inoculation (dpi). Viral genes were detected in 26 out of 28 air samples from both horses at 1–13 dpi by real-time RT-PCR. A rapid molecular test kit based on real-time RT-PCR detected viral genes in 23 air samples from one horse at 1–9 and 12 dpi, and from the other at 1–13 dpi. These findings confirm that horses infected with EIV shed the virus into the air. Air sampling is safe for humans and horses and avoids the potential for injury when nasopharyngeal swabs need to be collected from untrained or aggressive horses. EIV RNA was detected in the air samples by using real-time RT-PCR or the rapid molecular test kit before the horses showed clinical signs. Thus, air samplers can detect EIV RNA as early as possible through routine testing in locations such as quarantine facilities.

马流感病毒（EIV）可通过吸入雾化飞沫、直接接触和被污染的污染物传播。然而，据我们所知，没有从感染马匹周围的空气中恢复EIV的报告。在这里，我们使用空气采样器评估了EIV是否可以从实验感染马的马厩空气中恢复。此外，我们研究了反应时间小于30 min的快速分子检测试剂盒是否可以从空气样品中检测出EIV，以用于势场应用。分别饲养的两匹马实验性地感染了EIV。每天使用空气采样器采集空气样本，直至接种后13 d。通过实时RT-PCR，在每小时1-13 dpi的28个空气样本中检测到26个病毒基因。基于实时RT-PCR的快速分子检测试剂盒检测了一匹马在1-9和12 dpi以及另一匹马在1-13 dpi的23份空气样本中的病毒基因。这些发现证实，感染了EIV的马将病毒释放到空气中。空气取样对人和马都是安全的，当需要从未经训练或具有攻击性的马身上收集鼻咽拭子时，可避免潜在的伤害。在马出现临床症状前，采用实时RT-PCR或快速分子检测试剂盒检测空气样本中的EIV RNA。因此，空气采样器可以通过在检疫设施等地点的常规检测尽早检测到EIV RNA。

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引用次数: 0

RIPK3 activation of CaMKII triggers mitochondrial apoptosis in NIBV-infected renal tubular epithelial cells RIPK3激活CaMKII触发nibv感染肾小管上皮细胞线粒体凋亡

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2025-01-10 DOI: 10.1016/j.vetmic.2025.110375

Ying Li , Qiurong Qi , Yifei Chen, Mengbing Ding, Manzi Huang, Cheng Huang, Ping Liu, Xiaona Gao, Xiaoquan Guo, Zhanhong Zheng

The purpose of this study was to investigate whether RIPK3-mediated programmed cell death can promote the replication and transmission of renal infectious bronchitis virus in renal tubular epithelial cells. Primary renal tubular epithelial cells were extracted from 1 to 7 day old Hy-Line Brown chicks, cultured in vitro by type I collagenase digestion, and infected with 1MOI SX9 strain. Cell samples were collected at 12 hpi, 24 hpi, 36 hpi and 48 hpi for experimental exploration. Our results showed that NIBV infection could lead to programmed necrosis and mitochondrial apoptosis, and the expression levels of programmed necrosis-related genes TNFR1, TRADD, FADD, RIPK1, RIPK3 and MLKL increased significantly with the extension of infection time, the expression levels of mitochondrial apoptosis-related genes Cyt-C, APAF-1, Caspase-9 and Csapase-3 were significantly increased at 36 hpi. While, after 36 hpi of virus infection, apoptosis decreased and necrosis increased, and virus replication peaked. In order to further explore the effect of necroptosis on the amplification of renal infectious virus, the RIPK3 was inhibited at 36 hpi. Inhibition of necroptosis could reduce viral replication and cell death, programmed necrosis occurred in the cells, and cell membrane perforation led to virus diffusion and replication. NIBV-induced necroptosis depends on RIPK3, and RIPK3 activates CAMKII and interacts to cause abnormal opening of mitochondrial membrane permeability transition pore, promotes Ca^2 + influx into mitochondria, initiates mitochondrial apoptosis. While, inhibition of RIPK3 significantly inhibited the programmed necrosis of cells caused by NIBV infection, making excessive necrosis into moderate necrosis, thereby inhibiting the replication of the virus in cells.

本研究旨在探讨ripk3介导的程序性细胞死亡是否能促进肾传染性支气管炎病毒在肾小管上皮细胞中的复制和传播。从1 ~ 7日龄海兰褐鸡中提取原代肾小管上皮细胞，采用I型胶原酶消化法体外培养，并用1MOI SX9菌株感染。分别在12hpi、24hpi、36hpi和48hpi处采集细胞样本进行实验探索。我们的研究结果显示，NIBV感染可导致程序性坏死和线粒体凋亡，随着感染时间的延长，程序性坏死相关基因TNFR1、TRADD、FADD、RIPK1、RIPK3和MLKL的表达水平显著升高，线粒体凋亡相关基因Cyt-C、APAF-1、Caspase-9和Csapase-3的表达水平在36 hpi时显著升高。病毒感染36hpi后，细胞凋亡减少，坏死增加，病毒复制达到高峰。为了进一步探讨坏死坏死对肾感染性病毒扩增的影响，我们在36 hpi时抑制RIPK3。抑制坏死下垂可减少病毒复制和细胞死亡，细胞发生程序性坏死，细胞膜穿孔导致病毒扩散和复制。nibv诱导的坏死坏死依赖于RIPK3， RIPK3激活CAMKII并相互作用导致线粒体膜通透性过渡孔异常打开，促进Ca2 +内流线粒体，引发线粒体凋亡。而抑制RIPK3可显著抑制NIBV感染引起的细胞程序性坏死，使过度坏死变为中度坏死，从而抑制病毒在细胞中的复制。

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引用次数: 0