Veterinary microbiology最新文献_第3页

Mycoplasma synoviae surface-located elongation factor G functions as a cytoadhesin to promote adhesion to synovial sheath cells through binding to vimentin 滑膜支原体表面定位伸长因子 G 作为一种细胞粘附素，通过与波形蛋白结合促进与滑膜鞘细胞的粘附。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-11-01 DOI: 10.1016/j.vetmic.2024.110275

Bin Xu , Yu Sun , Shu Wang , Weiping Yao , Qing Wang , Ting Yuan , Sunting Ma , Xiaoli Wang , Lixin Lyu , Yanfei Yu , Xiaofei Zhang , Guoqing Shao , Wei Ouyang , Qiyan Xiong , Zhixin Feng

Mycoplasma synoviae infection has caused serious economic losses to the poultry industry worldwide. The molecular mechanism by which M. synoviae colonizes the synovium and induces synovitis is unclear. In this study, desthiobiotin pull-down and liquid chromatography-tandem mass spectrometry analyses were used to screen M. synoviae membrane proteins that bind the membrane proteins of synovial sheath cells (SSCs). Among the 128 screened proteins, elongation factor G (EF-G) of M. synoviae was identified as a surface-located protein using colony blotting and dual fluorescence analyses. The immunogenicity of EF-G was confirmed by the preparation of a rabbit polyclonal antibody. EF-G was identified as a cytoadhesin that directly binds to SSCs using indirect immunofluorescence assay and ELISA plate binding assay. In addition, antibody adhesion inhibition and protein adhesion inhibition demonstrated that EF-G could significantly promote the adhesion of M. synoviae to SSCs. Co-IP, GST pull-down, bacterial two-hybridization, and ELISA plate binding assays were performed to demonstrate the binding of EF-G and vimentin in vivo and in vitro. Antibody adhesion inhibition, protein adhesion inhibition, and siRNA interference adhesion inhibition assays demonstrated that vimentin significantly affected M. synoviae adhesion to SSCs. These studies indicate that two interacting proteins, EF-G, a novel cytoadhesin, and vimentin, an important cell surface receptor, play important roles in the adhesion of M. synoviae to SSCs, laying a foundation for subsequent studies on the mechanism of M. synoviae-induced synovitis and providing meaningful targets for screening target drugs against M. synoviae infection.

滑膜支原体感染给全球家禽业造成了严重的经济损失。滑膜支原体定植于滑膜并诱发滑膜炎的分子机制尚不清楚。本研究采用脱硫生物素牵引和液相色谱-串联质谱分析法筛选了与滑膜鞘细胞（SSCs）膜蛋白结合的滑膜杆菌膜蛋白。在筛选出的128种蛋白质中，通过菌落印迹和双荧光分析，确定了滑膜鞘膜杆菌的伸长因子G（EF-G）是一种表面定位蛋白。通过制备兔多克隆抗体证实了 EF-G 的免疫原性。通过间接免疫荧光测定和酶联免疫吸附板结合测定，确定了 EF-G 是一种可直接与 SSCs 结合的细胞粘附素。此外，抗体粘附抑制试验和蛋白粘附抑制试验表明，EF-G 能显著促进滑膜霉与 SSCs 的粘附。为了证明 EF-G 与波形蛋白在体内和体外的结合，研究人员进行了 Co-IP、GST pull-down、细菌双杂交和 ELISA 平板结合试验。抗体粘附抑制、蛋白粘附抑制和 siRNA 干扰粘附抑制试验表明，波形蛋白会显著影响滑膜霉菌对 SSCs 的粘附。这些研究表明，新型细胞粘附蛋白EF-G和重要的细胞表面受体vimentin这两种相互作用蛋白在滑膜霉菌粘附SSCs过程中发挥了重要作用，为后续研究滑膜霉菌诱发滑膜炎的机制奠定了基础，并为筛选抗滑膜霉菌感染的靶向药物提供了有意义的靶点。

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引用次数: 0

Antiviral role of cholesterol 25-hydroxylase in inhibiting porcine circovirus 3 replication 胆固醇 25- 羟化酶在抑制猪圆环病毒 3 复制中的抗病毒作用。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-22 DOI: 10.1016/j.vetmic.2024.110284

Baoge Zhang, Lumen Chao, Zhendong Wang, Hao Yu, Yufeng Li

Cholesterol 25-hydroxylase (CH25H) has significant antiviral effects through the production of 25-hydroxycholesterol (25HC). In this study, we investigated the effects of CH25H, its catalytic product 25HC, and its catalytic mutant lacking hydroxylase activity (CH25H-M) on porcine circovirus 3 (PCV3) replication. By transfecting PCV3 persistently infected PK-15 cells with the pCAGGS-CH25H-Flag plasmid, the results demonstrated that overexpression of CH25H significantly inhibited PCV3 Cap protein expression, Cap mRNA levels, and viral titers in a dose-dependent manner. Moreover, its catalytic product 25HC inhibited PCV3 replication in PK-15 cells at concentrations below 10 µM without affecting cell viability. In contrast, knockdown of endogenous CH25H using small interfering RNA (siRNA) enhanced PCV3 replication, further confirming its antiviral role. Interestingly, the CH25H-M mutant also exhibited inhibitory effects on PCV3 replication, although the inhibition was much less effective compared with CH25H. In conclusion, CH25H plays a critical role in regulating PCV3 replication, and its antiviral effect is not entirely dependent on its enzymatic activity. These findings provide new insights into both the enzymatic and non-enzymatic antiviral mechanisms of CH25H and revealed some mechanistic immune evasion for PCV3.

胆固醇 25- 羟化酶（CH25H）通过产生 25- 羟基胆固醇（25HC）具有显著的抗病毒作用。本研究调查了 CH25H、其催化产物 25HC 及其缺乏羟化酶活性的催化突变体（CH25H-M）对猪圆环病毒 3（PCV3）复制的影响。通过用 pCAGGS-CH25H-Flag 质粒转染 PCV3 持续感染的 PK-15 细胞，结果表明过表达 CH25H 能显著抑制 PCV3 Cap 蛋白的表达、Cap mRNA 水平和病毒滴度，且呈剂量依赖性。此外，其催化产物 25HC 在浓度低于 10 µM 时可抑制 PCV3 在 PK-15 细胞中的复制，且不影响细胞活力。相反，使用小干扰 RNA（siRNA）敲除内源性 CH25H 会增强 PCV3 复制，进一步证实了它的抗病毒作用。有趣的是，CH25H-M 突变体对 PCV3 复制也有抑制作用，但与 CH25H 相比，抑制效果要差得多。总之，CH25H 在调节 PCV3 复制过程中起着关键作用，其抗病毒作用并不完全依赖于其酶活性。这些发现为研究 CH25H 的酶和非酶抗病毒机制提供了新的视角，并揭示了 PCV3 的一些免疫逃避机制。

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引用次数: 0

Re-emergence of Bagaza virus in wild birds from southern Spain 西班牙南部野生鸟类中再次出现巴加扎病毒。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-18 DOI: 10.1016/j.vetmic.2024.110279

Moisés Gonzálvez , David Cano-Terriza , Úrsula Höfle , Félix Gómez-Guillamón , Cristina Cano-Gómez , Irene Zorrilla , Montserrat Agüero , Remigio Martínez , Ignacio García-Bocanegra

Bagaza virus (BAGV; Orthoflavivirus bagazaense) is an emerging vector-borne flavivirus affecting avian species with severe implications for animal health, and whose zoonotic potential has also been suggested. The aim of the present study was to monitor the epidemic outbreak of BAGV in wild birds from Spain in 2021. BAGV cases were confirmed in game bird species, including red-legged partridges (Alectoris rufa) and common pheasants (Phasianus colchicus) from 24 hunting areas. Suspected cases (clinical signs and/or mortality compatible with BAGV infection but without molecular confirmation) were also detected in 11 additional hunting areas. The outbreaks showed a seasonality mainly restricted to July and August. Estimated morbidity [25.8 % (95 %CI: 11.3–40.3) in partridges and 8.7 % (95 %CI: 0.0–18.0) in pheasants] and mortality rates [27.3 % (95 %CI: 12.5–42.1) in partridges and 13.0 % (95 %CI: 1.9–24.1) in pheasants] were found in the affected hunting areas. In addition, 215 non-game birds belonging to 46 different species were sampled by passive surveillance upon admittance to rehabilitation centres during 2021. BAGV infection was detected for the first time in green woodpecker (Picus viridis), spoonbill (Platalea leucorodia), white stork (Ciconia ciconia) and cynereous vulture (Aegypius monachus), expanding the host range of this emerging pathogen. In contrast to other species, game birds showed distinct BAGV related lesions, primarily myocarditis and encephalitis in addition to inflammatory infiltrates and necrosis in the liver and kidney. Molecular analyses revealed a homology of 97.4–98.0 % and 92.5–92.7 % between the BAGV sequences obtained in the present study (492 bp) and those isolated in 2010 and 2019 in southern Spain, respectively. These results allow to hypothesise about the likely silent and endemic circulation of BAGV since 2010 in this European region, although repeated virus reintroduction from neighbouring territories cannot be ruled out. Our findings evidence the sanitary, ecological and conservation implications of the re-emerged BAGV for wild birds, also emphasising the need to increase surveillance for monitoring and early detection of flavivirus dynamics in high-risk areas.

巴加扎病毒（BAGV；Orthoflavivirus bagazaense）是一种新出现的影响禽类物种的病媒黄病毒，对动物健康有严重影响，也有研究认为它有可能成为人畜共患病。本研究的目的是监测 2021 年在西班牙野鸟中爆发的 BAGV 流行病。24 个狩猎区的野禽物种（包括红腿鹧鸪（Alectoris rufa）和普通雉鸡（Phasianus colchicus））均确诊了 BAGV 病例。在另外 11 个狩猎区也发现了疑似病例（临床症状和/或死亡率与 BAGV 感染相符，但未经分子确认）。疫情主要集中在 7 月和 8 月。在受影响的狩猎区发现了估计发病率[鹧鸪为 25.8 %（95 %CI：11.3-40.3），雉鸡为 8.7 %（95 %CI：0.0-18.0）]和死亡率[鹧鸪为 27.3 %（95 %CI：12.5-42.1），雉鸡为 13.0 %（95 %CI：1.9-24.1）]。此外，在 2021 年期间，通过被动监测，对进入康复中心的 46 个不同物种的 215 只非野禽进行了采样。在绿啄木鸟（Picus viridis）、琵鹭（Platalea leucorodia）、白鹳（Ciconia ciconia）和秃鹫（Aegypius monachus）中首次检测到BAGV感染，扩大了这一新兴病原体的宿主范围。与其他物种不同的是，野禽表现出明显的 BAGV 相关病变，主要是心肌炎和脑炎，此外还有肝脏和肾脏的炎症浸润和坏死。分子分析表明，本研究中获得的 BAGV 序列（492 bp）与 2010 年和 2019 年在西班牙南部分离到的 BAGV 序列之间的同源性分别为 97.4-98.0 % 和 92.5-92.7 %。根据这些结果，我们可以推测自 2010 年以来，BAGV 病毒很可能在这一欧洲地区静默流行，但也不能排除病毒从邻近地区再次传入的可能性。我们的研究结果证明了再次出现的 BAGV 对野生鸟类的卫生、生态和保护的影响，同时也强调了在高风险地区加强监测和早期发现黄病毒动态的必要性。

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引用次数: 0

Recombinant avian-derived antiviral proteins cIFITM1, cIFITM3, and cViperin as effective adjuvants in inactivated H9N2 subtype avian influenza vaccines 重组禽源性抗病毒蛋白 cIFITM1、cIFITM3 和 cViperin 作为 H9N2 亚型禽流感灭活疫苗的有效佐剂。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-18 DOI: 10.1016/j.vetmic.2024.110277

Shihong Yan , Yikai Chen , Jin Lin , Huimin Chen , Chenqi Hu , Hongyang Liu , Hongxiu Diao , Shasha Liu , Ji-Long Chen

Vaccine adjuvants, serving as non-specific immune enhancers, play a pivotal role in the immunoprevention and control of animal diseases. This study utilized prokaryotic expression systems to express and purify chicken-derived cIFITM1, cIFITM3, and cViperin, which were then formulated as adjuvants with H9N2 avian influenza virus antigens to create inactivated vaccines. These vaccines were administered to SPF chickens to investigate their immunopotentiating functions. Additionally, the proteins were assessed for their ability to act as standalone immune enhancers. The results demonstrated that cIFITM1, cIFITM3, and cViperin significantly elevated serum hemagglutination inhibition (HI) antibody titers. Notably, when used individually, these proteins markedly enhanced the antiviral capabilities of challenged chickens, leading to alleviated clinical symptoms, reduced tracheal virus replication, diminished virus shedding, and lessened histopathological damage, with cIFITM1 exhibiting the most pronounced effect. Furthermore, the protective efficacy of two H9N2 recombinant virus inactivated vaccines supplemented with cIFITM1 adjuvant was validated, achieving a 100 % vaccine protection efficiency. In conclusion, cIFITM1, cIFITM3, and cViperin as adjuvants for influenza vaccines effectively inhibit virus replication and shedding, highlighting their significant potential in influenza prevention and control.

疫苗佐剂作为非特异性免疫增强剂，在动物疾病的免疫预防和控制中发挥着举足轻重的作用。本研究利用原核表达系统表达和纯化鸡源性 cIFITM1、cIFITM3 和 cViperin，然后将其作为佐剂与 H9N2 禽流感病毒抗原配制成灭活疫苗。将这些疫苗注射给 SPF 鸡，以研究它们的免疫增强功能。此外，还评估了这些蛋白质作为独立免疫增强剂的能力。结果表明，cIFITM1、cIFITM3 和 cViperin 能显著提高血清血凝抑制（HI）抗体滴度。值得注意的是，当单独使用这些蛋白时，它们能明显增强受挑战鸡的抗病毒能力，从而缓解临床症状、减少气管病毒复制、减少病毒脱落和减轻组织病理学损伤，其中 cIFITM1 的效果最为明显。此外，两种添加了 cIFITM1 佐剂的 H9N2 重组病毒灭活疫苗的保护效力也得到了验证，疫苗保护效率达到了 100%。总之，cIFITM1、cIFITM3 和 cViperin 作为流感疫苗佐剂可有效抑制病毒复制和脱落，在流感防控中具有重要潜力。

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引用次数: 0

Antimicrobial susceptibility of Clostridium botulinum group III field strains isolated in Europe from animal outbreaks 欧洲从动物疫情中分离出的第三类肉毒梭状芽孢杆菌野外菌株的抗菌药敏感性。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-16 DOI: 10.1016/j.vetmic.2024.110281

Benedetta Cordioli , Alessia Rizzardi , Angela Guolo , Tiziana Ferro , Cosetta Bacchin , Manuel Garbuio , Fabrizio Anniballi , Paola De Santis , Miriam Koene , Caroline Le Maréchal , Hanna Skarin , Christian Seyboldt , Luca Bano

Neurotoxins produced by Clostridium (C.) botulinum group III are responsible for the majority of botulism outbreaks occurring in animals and in this study we report the drug susceptibility of 71 field strains. The minimum inhibitory concentration (MIC) of 13 antimicrobials was established through the agar dilution method. The MIC₅₀ matched or differed for one or two dilutions from MIC₉₀ of the same antimicrobial, showing a unimodal distribution of the MIC values, irrespective of the geographical origin, the animal source and the toxinotype of the strain. Beta-lactams and rifampin showed the lowest MIC values, while gentamicin, polymyxin B and sulfamethoxazole showed the highest MICs. As for similar studies conducted in human botulism, the results could be helpful to avoid the administration of antimicrobials that could worsen the health condition of the affected animals and to develop selective media for the isolation of these fastidious anaerobes. Indeed, the isolation of the strain from affected animals and from environmental samples is important to perform epidemiological studies based on the genetic characterization and to produce tailor-made vaccines.

肉毒梭状芽孢杆菌（C. botulinum）III群产生的神经毒素是导致动物肉毒中毒事件爆发的主要原因，在这项研究中，我们报告了71株野外菌株对药物的敏感性。通过琼脂稀释法确定了 13 种抗菌药物的最低抑菌浓度（MIC）。在一个或两个稀释度中，MIC50 与同一种抗菌素的 MIC90 相匹配或存在差异，显示出 MIC 值的单峰分布，与菌株的地理来源、动物来源和毒素类型无关。β-内酰胺类和利福平的 MIC 值最低，而庆大霉素、多粘菌素 B 和磺胺甲噁唑的 MIC 值最高。至于在人类肉毒中毒中进行的类似研究，其结果可能有助于避免使用可能会恶化患病动物健康状况的抗菌药，并有助于开发用于分离这些苛刻厌氧菌的选择性培养基。事实上，从患病动物和环境样本中分离菌株对于根据基因特征进行流行病学研究和生产定制疫苗非常重要。

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引用次数: 0

Transmission of carbapenem-resistant enterobacterales producing NDM-5 during the broiler breeding process in China 中国肉鸡养殖过程中产生NDM-5的耐碳青霉烯类肠杆菌的传播

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-16 DOI: 10.1016/j.vetmic.2024.110282

Ke Wu , Yuanyuan Zhang , Wei Xu , Xiaolong Lin , Cui Li , Juan Wang , Ruichao Li , Yizhi Tang , Changwei Lei , Hongning Wang

This study conducted a four-month monitoring of carbapenem resistance in a broiler breeding farm in China. A total of 185 carbapenem-resistant bacterial isolates were obtained from 2298 cloacal swabs from broiler breeders and their offspring within a production cycle. The detection rate of carbapenem-resistant isolates was higher during the brooding period. Whole-genome sequencing (WGS) was performed on 133 isolates based on sampling stages, including 113 carbapenem-resistant Enterobacterales (CRE) isolates and 20 Stenotrophomonas pavanii isolates, which have intrinsic resistance to carbapenems. A total of 69 antibiotic resistance genes (ARGs), including bla_NDM-1, mcr-1, and bla_NDM-5, were identified among the sequenced CRE isolates. Notably, bla_NDM-5 (92.0 %, 104/113) was the primary contributor to carbapenem resistance. CRE isolates from the same breeding stage exhibited close genomic relationships, and the bla_NDM-5 genes were observed in similar genetic backgrounds, indicating the transmission of CRE strains and bla_NDM-5 during the broiler breeding process. No CRE was isolated from 0 d broiler offspring, suggesting that broiler breeders were not the direct source of CRE in their offspring. Tracing the feeding process revealed that brooder and rearing houses were likely key factors in the cross-transmission of CRE between broiler breeders and their offspring. CRE pose a significant threat to public health and food safety. China is one of the world's leading poultry producing and consuming countries. This study provided insights into the epidemiological trends and key transmission nodes of carbapenem resistance and CRE within the broiler breeding process, which could help the control of antibiotic resistance and bacterial infections in the broiler industry.

本研究对中国一家肉种鸡养殖场的碳青霉烯类耐药性进行了为期四个月的监测。在一个生产周期内，从肉用种鸡及其后代的 2298 份泄殖腔拭子中共分离出 185 株耐碳青霉烯类细菌。在育雏期，耐碳青霉烯类细菌分离株的检出率较高。根据采样阶段，对 133 个分离物进行了全基因组测序（WGS），其中包括 113 个耐碳青霉烯类肠杆菌（CRE）分离物和 20 个对碳青霉烯类有内在耐药性的铺满性苯并酵母菌（Stenotrophomonas pavanii）分离物。在测序的 CRE 分离物中共鉴定出 69 个抗生素耐药基因（ARGs），包括 blaNDM-1、mcr-1 和 blaNDM-5。值得注意的是，blaNDM-5（92.0%，104/113）是造成碳青霉烯类耐药性的主要原因。来自同一育种阶段的 CRE 分离物表现出密切的基因组关系，并且在相似的遗传背景中观察到 blaNDM-5 基因，这表明 CRE 菌株和 blaNDM-5 在肉鸡育种过程中存在传播。从 0 d 后代肉鸡中未分离到 CRE，表明肉种鸡不是后代中 CRE 的直接来源。对饲养过程的追踪表明，育雏器和育雏舍可能是肉种鸡及其后代之间交叉传播 CRE 的关键因素。CRE对公众健康和食品安全构成重大威胁。中国是世界上主要的家禽生产国和消费国之一。本研究深入探讨了肉鸡养殖过程中碳青霉烯类耐药性和CRE的流行趋势和关键传播节点，有助于控制肉鸡行业的抗生素耐药性和细菌感染。

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引用次数: 0

Epidemiological investigation of equine rotavirus B outbreaks in horses in central Kentucky 肯塔基州中部马匹轮状病毒 B 爆发的流行病学调查。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-16 DOI: 10.1016/j.vetmic.2024.110278

Chithra C. Sreenivasan , Ahsan Naveed , Tirth Uprety, Shalini Soni, Olivia Jacob, Emma Adam, Dan Wang, Feng Li

Using metagenomic sequencing we identified equine rotavirus group B (ERVB) of ruminant origin in foal diarrhea outbreaks in the 2021 foaling season. To further investigate ERVB occurrence and determine its environmental stability, we collected mare and foal fecal samples from different farms in Central Kentucky during the 2022 foaling season. The RT-qPCR-based analyses showed that ERVB genome was detected in 16.67 % (42/252) of surveyed mare samples and 26.56 % (34/128) of foal samples. Furthermore, 94.12 % (16/17) of collected soil samples and 100 % (13/13) of water samples obtained from the ERVB-positive farm premises also tested weakly positive. In addition, ERVB genome fragments were detected in 58.33 % (7/12) of indoor samples collected from the equipment/barn/hospital wards during the outbreak period. Finally, the seroprevalence study showed 87 % (113/130) of surveyed horse serum samples were positive for ERVB antibodies. Despite unsuccessful attempts in ERVB cultivation, phylogenetic analyses showed that fecal ERVB strains representing 2022 and 2023 foal diarrhea outbreaks, like 2021 strains, were more closely related to ruminant rotavirus B than other viruses. Further sequence analyses revealed that none of the three viral capsid proteins, the primary targets of virus-neutralizing antibodies, exhibited notable mutations among ERVB strains circulated over the past three years. Our data demonstrated that ERVB was widespread in horses on affected farms with extreme stability in the farm environment. These findings continue to support the need for future surveillance of ERVB in horses and the surrounding environment, and the development of effective countermeasures to protect horses against this new viral disease.

通过元基因组测序，我们在 2021 年产驹季节爆发的马驹腹泻中发现了反刍动物来源的马轮状病毒 B 组（ERVB）。为了进一步调查 ERVB 的发生情况并确定其环境稳定性，我们在 2022 年的产驹季节收集了肯塔基州中部不同农场的母马和马驹粪便样本。基于 RT-qPCR 的分析表明，在调查的母马样本中有 16.67 %（42/252）和马驹样本中有 26.56 %（34/128）检测到 ERVB 基因组。此外，从ERVB阳性农场采集的94.12%（16/17）的土壤样本和100%（13/13）的水样本也检测出弱阳性。此外，在疫情爆发期间从设备/谷仓/医院病房采集的室内样本中，有 58.33 %（7/12）的样本检测到 ERVB 基因组片段。最后，血清流行率研究显示，87%（113/130）的马血清样本中ERVB抗体呈阳性。尽管ERVB的培养尝试并不成功，但系统进化分析表明，代表2022年和2023年马驹腹泻疫情的粪便ERVB毒株与2021年的毒株一样，与反刍轮状病毒B的亲缘关系比其他病毒更近。进一步的序列分析表明，在过去三年中流行的ERVB毒株中，病毒中和抗体的主要靶标--三种病毒外壳蛋白都没有出现明显的变异。我们的数据表明，ERVB 在受影响马场的马匹中普遍存在，且在马场环境中极为稳定。这些发现继续支持了今后对马匹和周围环境中的 ERVB 进行监测的必要性，以及开发有效对策保护马匹免受这种新型病毒性疾病侵害的必要性。

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引用次数: 0

A novel neuro-attenuated vaccine candidate with excellent safety and protective efficacy against highly virulent Feline Herpesvirus-1 一种新型神经减毒候选疫苗，对高致病性猫疱疹病毒-1 具有极佳的安全性和保护效力。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-15 DOI: 10.1016/j.vetmic.2024.110276

Mingyu Qi , Mengfang Yang , Ruxue Luo , Lingying Fang , Yixi Chen , Jianuo Gao , Zhe Jiao , Yuejun Shi , Guiqing Peng

Feline herpesvirus 1 (FHV-1) is a major pathogen responsible for respiratory, ocular and nervous system symptoms in felines. FHV-1 can remain latenct in ganglia and is difficult to eliminate completely with drug treatment. Currently, commercially FHV-1 vaccines are not sufficiently effective and provide only limited durations of protection. To enhance vaccine efficacy and reduce latent virus in tissues, two gene deletion mutants of FHV-1 conveyed excellent proliferation ability, genetic stability and attenuated FHV-1 virulence were constructed by CRISPR/Cas9-mediated homologous recombination, designated as FHV-△US3 and FHV-△UL50. Recombinant FHV-1 induce stronger cellular and humoral immune responses, as well as better protective effects than those of commercial vaccines. Notably, FHV-△US3 and FHV-△UL50 reveal neuro-attenuated, as viral residue in the trigeminal ganglia are significantly reduced. The knockout of the UL50 gene in FHV-1 has not been previously reported. In this study, we aimed to evaluate the safety and immunogenicity of FHV-△UL50, highlighting its potential as a novel neuroattenuated vaccine candidate.

猫疱疹病毒 1 (FHV-1) 是导致猫科动物出现呼吸道、眼部和神经系统症状的主要病原体。FHV-1 可潜伏在神经节中，很难通过药物治疗彻底清除。目前，市售的 FHV-1 疫苗效果不佳，只能提供有限的保护期。为了提高疫苗疗效并减少组织中的潜伏病毒，我们通过 CRISPR/Cas9 介导的同源重组技术构建了两个基因缺失的 FHV-1 突变体，这两个突变体分别被命名为 FHV-△US3 和 FHV-△UL50，它们具有优异的增殖能力、遗传稳定性和减毒能力。与商业疫苗相比，重组 FHV-1 能诱导更强的细胞和体液免疫反应，以及更好的保护效果。值得注意的是，FHV-△US3 和 FHV-△UL50 揭示了神经减毒作用，三叉神经节中的病毒残留明显减少。之前还没有关于 FHV-1 中 UL50 基因敲除的报道。在这项研究中，我们旨在评估FHV-△UL50的安全性和免疫原性，突出其作为新型神经减毒疫苗候选物的潜力。

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引用次数: 0

IFN-β contributes to astrocyte activation in the brain following coronavirus PHEV infection independent on peripheral immunity IFN-β有助于冠状病毒 PHEV 感染后大脑星形胶质细胞的活化，与外周免疫无关。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-15 DOI: 10.1016/j.vetmic.2024.110280

Junchao Shi , Qianhan Jia , Yanfang Yu , Jing Zhang , Zi Li , Wenqi He

Porcine hemagglutinating encephalomyelitis (PHE), caused by a betacoronavirus named porcine hemagglutinating encephalomyelitis virus (PHEV), is a highly fatal disease of pigs characterized by nonsuppurative encephalitis. Activation of astrocytes is a hallmark of viral encephalomyelitis; however, the mechanism of PHEV-induced astrocyte activation is currently unknown. Based on mouse model, we show that PHEV infection led to astrogliosis in mouse brain and brain slice cultures (BSCs), as indicated by increased expression of glial fibrillary acidic protein (GFAP). PHEV can neither infect nor activate primary astrocytes in vitro, indicating that activation of astrocytes maybe mediated by factors secreted from viral infected neurons but not by direct viral infection of astrocytes. PHEV infection results in increased interferon (IFN) response in later stage, we thereafter focused on whether IFN-β can activate astrocytes after PHEV infection similar to other neurotropic viruses. IFN-β treatment resulted in both the upregulation of GFAP and activation-associated cytokines/chemokines in mouse primary astrocytes. Furthermore, the addition of IFN-β neutralization antibody prevented PHEV-infected mouse brain tissue homogenate from activating astrocytes. Taken together, IFN-β triggers the activation of astrocytes in the central nervous system (CNS) following PHEV infection. Further understanding of the role of activated astrocytes during PHEV infection may provide new insights for treatment this disease.

猪血凝性脑脊髓炎（PHE）是由一种名为猪血凝性脑脊髓炎病毒（PHEV）的betacoronavirus引起的，是一种高度致命的猪疾病，以非化脓性脑炎为特征。星形胶质细胞活化是病毒性脑脊髓炎的特征之一；然而，PHEV 诱导星形胶质细胞活化的机制目前尚不清楚。基于小鼠模型，我们发现 PHEV 感染会导致小鼠大脑和脑片培养物（BSCs）中的星形胶质细胞增生，表现为胶质纤维酸性蛋白（GFAP）的表达增加。PHEV 既不能感染也不能激活体外原发性星形胶质细胞，这表明星形胶质细胞的激活可能是由病毒感染的神经元分泌的因子介导的，而不是由病毒直接感染星形胶质细胞介导的。PHEV 感染后期会导致干扰素（IFN）反应增强，因此我们关注 PHEV 感染后 IFN-β 是否能像其他神经毒性病毒一样激活星形胶质细胞。IFN-β 处理可导致小鼠原代星形胶质细胞的 GFAP 和活化相关细胞因子/趋化因子上调。此外，添加 IFN-β 中和抗体可防止 PHEV 感染的小鼠脑组织匀浆激活星形胶质细胞。综上所述，PHEV 感染后，IFN-β 会引发中枢神经系统（CNS）中星形胶质细胞的活化。进一步了解 PHEV 感染期间活化星形胶质细胞的作用可能会为治疗这种疾病提供新的见解。

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引用次数: 0

The prevalence and shedding of porcine epidemic diarrhea virus in intensive swine farms of China from 2022 to 2023 2022 至 2023 年中国集约化猪场猪流行性腹泻病毒的流行和脱落情况

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-11 DOI: 10.1016/j.vetmic.2024.110273

Yuetao Xu , Meng Sun , Zhunxuan Wang , Xiaowen Li , Yijun Du , Ping Jiang

Porcine epidemic diarrhea has emerged as a significant threat to the global swine industry. The shedding and exposure status of porcine epidemic diarrhea virus (PEDV) in intensive farms is not completely understood. In this study, a total of 56,598 clinical samples collected from 256 intensive pig farms in 20 provinces in China from 2022 to 2023, were evaluated for PEDV using quantitative real-time PCR. The overall PEDV prevalence was 11.78 % and 28.45 % at the sample and farm levels, respectively, which are relatively high in Northern China and the fourth and first quarter of the year. The PEDV-positive rates and viral loads in suckling piglet herds were higher than those in growing-finishing pigs and multiparous sows. Meanwhile, 15.61 % of pig pens, 9.51 % of corridors, 9.4 % of office areas, 9.23 % of production personnel, and 8.33 % of pig cart driver samples were positive for PEDV, indicating potential biosafety gaps in intensive pig farms. In addition, 93.41 % of inguinal lymph node tissue samples contained viral nucleic acids, revealing a possible persistent infection of PEDV in pig herds. Our study presents the first report of the large-scale detection of PEDV in intensive pig farms, which constitutes indirect evidence of virus circulation in pig herds. This study provides valuable data for preventing and controlling PEDV infection in the future.

猪流行性腹泻已成为全球养猪业的一大威胁。猪流行性腹泻病毒（PEDV）在集约化养猪场的脱落和暴露状况尚不完全清楚。本研究采用实时定量 PCR 技术，对 2022 年至 2023 年期间从中国 20 个省的 256 个集约化猪场采集的 56598 份临床样本进行了 PEDV 评估。在样本和猪场层面，PEDV 的总体流行率分别为 11.78 % 和 28.45 %，在华北地区以及每年的第四季度和第一季度相对较高。乳仔猪群的 PEDV 阳性率和病毒载量均高于生长育成猪和多胎母猪。同时，15.61%的猪栏、9.51%的走廊、9.4%的办公区、9.23%的生产人员和 8.33%的猪车司机样本对 PEDV 呈阳性，这表明集约化猪场可能存在生物安全漏洞。此外，93.41% 的腹股沟淋巴结组织样本含有病毒核酸，揭示了 PEDV 可能在猪群中持续感染。我们的研究首次报告了在集约化养猪场大规模检测到 PEDV 的情况，这构成了猪群中病毒传播的间接证据。这项研究为今后预防和控制 PEDV 感染提供了宝贵的数据。

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引用次数: 0