Veterinary microbiology最新文献_第5页

Thioredoxin C of Streptococcus suis serotype 2 contributes to virulence by inducing antioxidative stress and inhibiting autophagy via the MSR1/PI3K-Akt-mTOR pathway in macrophages 猪链球菌血清型 2 的硫氧还蛋白 C 通过 MSR1/PI3K-Akt-mTOR 通路在巨噬细胞中诱导抗氧化压力和抑制自噬，从而增强毒力

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-24 DOI: 10.1016/j.vetmic.2024.110263

Chunxiao Ji , Yanying Pan , Bocheng Liu , Jianying Liu , Chijun Zhao , Zhuyuan Nie , Simeng Liao , Guangwei Kuang , Xin Wu , Quan Liu , Jie Ning , Yulong Tang , Lihua Fang

The thioredoxin (Trx) system plays a vital role in protecting against oxidative stress and ensures correct disulfide bonding to maintain protein function. Our previous research demonstrated that TrxA of Streptococcus suis Serotype 2 (SS2), a clinical strain from the lung of a diseased pig, contributes to virulence but is not involved in antioxidative stress. In this study, we identified another gene in the Trx family, TrxC, which encodes a protein of 104 amino acids with a CGDC active motif and 22.4 % amino acid sequence homology with TrxA. Unlike the TrxA, TrxC mutant strains were more susceptible to oxidative stresses induced by hydrogen peroxide and paraquat. In vitro experiments, the survival rate of the TrxC deletion mutant in RAW264.7 macrophages was only one-eighth of that of TrxA mutant strains. Transcriptome analysis revealed that autophagy-related genes were significantly upregulated in the TrxC mutant compared to those in the wild-type or TrxA mutant strains. Co-localization of LC3 puncta with TrxC was confirmed using laser confocal microscopy, and autophagy-related indicators were quantified using western blotting. Autophagy deficiency induced by ATG5 knockout significantly increased SS2 survival rate, especially in TrxC mutant strains. For the upstream signal regulation pathways, we found ΔTrxC strains regulate autophagy by activation of PI3K/Akt/mTOR signaling in RAW264.7 macrophages. In the Akt1-overexpressing cell line, ΔTrxC infection significantly decreased the autophagic response and promoted ΔTrxC mutant strain survival, while inhibition of Akt with MK2206 resulted in reduced ΔTrxC mutant strain survival and enhance the autophagic response. Furthermore, loss of TrxC increased the activity of MSR1, thereby inducing cellular autophagy and phagocytosis. Our data demonstrate that TrxC of SS2 contributes to virulence by inducing antioxidative stress and inhibits autophagy via the PI3K-Akt-mTOR pathway in macrophages, with MSR1 acting as a key factor in controlling infection.

硫代氧化酶（Trx）系统在抵御氧化应激和确保正确的二硫键以维持蛋白质功能方面发挥着重要作用。我们之前的研究表明，猪链球菌血清型 2（SS2）（一种来自病猪肺部的临床菌株）的 TrxA 有助于提高毒力，但并不参与抗氧化应激。在这项研究中，我们发现了 Trx 家族的另一个基因 TrxC，它编码一种 104 个氨基酸的蛋白质，具有 CGDC 活性基序，与 TrxA 的氨基酸序列同源性为 22.4%。与 TrxA 不同，TrxC 突变菌株更容易受到过氧化氢和百草枯诱导的氧化胁迫。在体外实验中，TrxC缺失突变株在 RAW264.7 巨噬细胞中的存活率仅为 TrxA 突变株的八分之一。转录组分析显示，与野生型或TrxA突变株相比，TrxC突变株的自噬相关基因明显上调。激光共聚焦显微镜证实了LC3点与TrxC的共定位，并利用Western印迹对自噬相关指标进行了定量。ATG5敲除诱导的自噬缺陷显著提高了SS2的存活率，尤其是在TrxC突变株中。在上游信号调控途径方面，我们发现ΔTrxC菌株通过激活RAW264.7巨噬细胞中的PI3K/Akt/mTOR信号来调控自噬。在Akt1表达缺失的细胞系中，ΔTrxC感染会显著降低自噬反应并促进ΔTrxC突变株存活，而用MK2206抑制Akt会降低ΔTrxC突变株存活并增强自噬反应。此外，TrxC 的缺失增加了 MSR1 的活性，从而诱导细胞自噬和吞噬。我们的数据表明，SS2的TrxC通过巨噬细胞中的PI3K-Akt-mTOR途径诱导抗氧化应激和抑制自噬，从而增强了病毒的毒力，而MSR1则是控制感染的关键因素。

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引用次数: 0

Development of a recombinant Lactobacillus plantarum oral vaccine expressing VP2 protein for preventing feline panleukopenia virus 开发表达 VP2 蛋白的重组植物乳杆菌口服疫苗以预防猫泛白细胞减少症病毒

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110257

Jiakang Li , Yue Zeng , Luying Li , Jiajia Peng , Quanhui Yan , Zijun Ye , Yan Zhang , Weihui Li , Longlong Cao , Dengyuan Zhou , QiuYan Li , Youhui Si , Shengbo Cao

Feline panleukopenia virus (FPV) represents a significant health threat to the kittens. While traditional vaccines administered via subcutaneous or intramuscular injection are effective, they can induce stress and adverse reactions. Moreover, unvaccinated kittens visiting veterinary clinics risk exposure to FPV, increasing their susceptibility to infection. Therefore, there is an urgent need for a safer, more gentle vaccination method with streamlined administration. In this study, we developed a recombinant L. plantarum NC8/VP2 expressing the VP2 protein of the prevalent Chinese FPV strain, FPV-251. Our results show that L. plantarum NC8/VP2 effectively colonizes the feline intestinal tract and induces high levels of neutralizing antibodies through oral administration. Kittens exhibited significant protection against FPV-251 infection and associated illnesses or fatalities after 30 days of continuous dosing. These results highlight the potential of recombinant L. plantarum NC8/VP2 as a novel oral vaccine for FPV, presenting a promising approach for disease prevention in domestic cats.

猫泛白细胞减少症病毒 (FPV) 对幼猫的健康构成严重威胁。虽然通过皮下注射或肌肉注射的传统疫苗很有效，但它们可能会引起应激反应和不良反应。此外，未接种疫苗的幼猫到兽医诊所就诊时有可能接触到 FPV，从而增加感染的几率。因此，我们迫切需要一种更安全、更温和、更简便的疫苗接种方法。在本研究中，我们开发了表达中国流行的 FPV 株 FPV-251 的 VP2 蛋白的重组植物乳杆菌 NC8/VP2。结果表明，植物乳杆菌 NC8/VP2 能有效定植于猫肠道，并通过口服诱导高水平的中和抗体。小猫在连续服药 30 天后，对 FPV-251 感染和相关疾病或死亡表现出明显的保护作用。这些结果凸显了重组植物乳杆菌 NC8/VP2 作为 FPV 新型口服疫苗的潜力，为家猫疾病预防提供了一种前景广阔的方法。

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引用次数: 0

Identification of three novel linear B-cell epitopes on VP7 of African horse sickness virus using monoclonal antibodies 利用单克隆抗体鉴定非洲马瘟病毒 VP7 上的三个新型线性 B 细胞表位

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110258

Xinbing Hu , Jing Xu , Xuanying Wang , Zhancheng Tian , Guiquan Guan , Jianxun Luo , Hong Yin , Junzheng Du

African horse sickness (AHS) is an acute and subacute infectious disease of equine species caused by the African horse sickness virus (AHSV). The VP7 of AHSV is a group-specific protein conserved in all serotypes and is an excellent candidate for the serological diagnosis and an AHS vaccine component. However, to date, B-cell epitopes on the AHSV VP7 recognized by humoral immune responses remain unclear. This study expressed the recombinant AHSV VP7 soluble in Escherichia coli and purified it for mouse immunization. Four monoclonal antibodies (mAbs) were screened and identified by hybridoma cell fusion, clonal purification, and immunological assays. The B-cell epitopes, recognized by monoclonal antibodies 4B5, 3G10, 3D7, and 4D6, were identified by a series of truncated overlapping peptides expressed as glutathione S-transferase (GST)-fusion proteins. The results revealed that 4B5 recognized the ¹²⁴VQTGRYAGA¹³² motif, 3G10 recognized the ¹⁴⁰RYYVPQGRT¹⁴⁸ motif, while 3D7 and 4D6 recognized the ²⁹²QPINPPIFP³⁰⁰ motif. Amino acid sequence alignment indicated that three novel B-cell epitopes were conserved among various AHSV serotypes but unconserved in other orbiviruses, such as the bluetongue and epidemic hemorrhagic disease viruses. This study informs on the antigenic epitopes of AHSV VP7, facilitating future investigations into the serological diagnosis method and epitope-based vaccines against AHSV.

非洲马瘟（AHS）是由非洲马瘟病毒（AHSV）引起的马科动物的一种急性和亚急性传染病。非洲马瘟病毒的 VP7 是一种在所有血清型中都保留下来的群体特异性蛋白，是血清学诊断和非洲马瘟疫苗成分的绝佳候选蛋白。然而，迄今为止，体液免疫反应识别的 AHSV VP7 上的 B 细胞表位仍不清楚。本研究在大肠杆菌中表达了可溶性重组 AHSV VP7，并将其纯化用于小鼠免疫。通过杂交瘤细胞融合、克隆纯化和免疫测定，筛选并鉴定了四种单克隆抗体（mAbs）。单克隆抗体 4B5、3G10、3D7 和 4D6 识别的 B 细胞表位是通过一系列截短的重叠肽鉴定的，这些肽表达为谷胱甘肽 S 转移酶（GST）融合蛋白。结果显示，4B5 识别 124VQTGRYAGA132 主题，3G10 识别 140RYYVPQGRT148 主题，而 3D7 和 4D6 识别 292QPINPPIFP300 主题。氨基酸序列比对表明，三种新型 B 细胞表位在 AHSV 不同血清型中是保守的，但在蓝舌病病毒和流行性出血病病毒等其他眼病毒中则是不保守的。这项研究为研究 AHSV VP7 的抗原表位提供了信息，有助于今后研究 AHSV 的血清学诊断方法和基于表位的疫苗。

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引用次数: 0

Longitudinal survey of hepatitis E virus in extensively raised pigs in Spain 西班牙广泛饲养的猪体内戊型肝炎病毒的纵向调查。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110256

Tomás Fajardo-Alonso , Ignacio García-Bocanegra , María A. Risalde , Antonio Rivero-Juárez , Saúl Jiménez-Ruiz , David Cano-Terriza , María Casares-Jiménez , Eduardo Laguna , Pelayo Acevedo , Mario Frías , Joaquín Vicente , Antonio Rivero , Javier Caballero-Gómez

Hepatitis E virus (HEV) is an emerging zoonotic virus of public health concern, of which pigs, wild boar and red deer are the main reservoirs. The European Food Safety Authority (EFSA) has recently prioritized the development of monitoring programs of HEV at different stages of the pig food chain, including outdoor pig farming. Pigs managed under these extensive production systems frequently share habitat and natural resources with wild boar and red deer during fattening stages and cross-species transmission of HEV among these species has previously been suggested. In this context, we aimed to (I) to evaluate the risk of HEV circulation within the production phases of extensively raised pigs and at the domestic-wildlife interface, and (II) to identify the genotypes circulating within these hosts. A total of 1452 pigs from seven different pig farms were longitudinally sampled during the breeding, rearing, and fattening production phases. In addition, 138 and 252 sympatric wild boar and red deer, respectively, were analysed. Anti-HEV antibodies were found in 1245 (85.7 %) out of the 1452 Iberian pigs sampled. The seroprevalence was 30.4 % in the breeding phase, 95.4 % in the rearing phase and 97.0 % in the fattening phase. Statistically significant differences (P < 0.05) were found among the three production phases. The seroprevalence was significantly higher (P < 0.001) in fattening pigs compared to those found in sympatric wild boar (31.9 %) and red deer (2.0 %). Three (1.0 %) out of the 293 serum pools analysed were positive for viral RNA. One of them was identified in pigs at the rearing phase (genotype 3 f) and two in wild boar (genotypes 3 f and 3 m). The high seroprevalence detected in extensively raised pigs, together with the detection of the zoonotic HEV-3 f and HEV-3 m subtypes in sympatric domestic and wild swine, highlights the risk of zoonotic transmission and the need to establish surveillance programs and control measures, particularly in breeding and rearing phase, in these epidemiological scenarios.

戊型肝炎病毒（HEV）是一种新出现的引起公共卫生关注的人畜共患病毒，猪、野猪和梅花鹿是其主要传播者。欧洲食品安全局（EFSA）最近优先考虑在猪食物链的不同阶段（包括户外养猪）制定 HEV 监测计划。在这些粗放型生产系统下管理的猪在育肥阶段经常与野猪和红鹿共享栖息地和自然资源，以前曾有人提出过 HEV 在这些物种间的跨物种传播。在这种情况下，我们的目标是：（I）评估 HEV 在广泛饲养的猪的生产阶段以及在家畜与野生动物交界处传播的风险；（II）确定在这些宿主中传播的基因型。在种猪、育成猪和育肥猪生产阶段，对来自 7 个不同猪场的 1452 头猪进行了纵向采样。此外，还分别对 138 头和 252 头同域野猪和赤鹿进行了分析。在采样的 1452 头伊比利亚猪中，有 1245 头（85.7%）发现了抗 HEV 抗体。血清流行率在育种阶段为 30.4%，在饲养阶段为 95.4%，在育肥阶段为 97.0%。三个生产阶段之间存在明显的统计学差异（P < 0.05）。与同域野猪（31.9 %）和赤鹿（2.0 %）相比，育肥猪的血清流行率明显更高（P < 0.001）。在分析的 293 份血清池中，有 3 份（1.0 %）病毒 RNA 呈阳性。其中一个在饲养阶段的猪（基因型 3 f）中发现，两个在野猪（基因型 3 f 和 3 m）中发现。在广泛饲养的猪中检测到的高血清流行率，以及在同域家猪和野猪中检测到的人畜共患的 HEV-3 f 和 HEV-3 m 亚型，突出表明了人畜共患传播的风险，以及在这些流行病学情况下建立监测计划和控制措施的必要性，特别是在繁殖和饲养阶段。

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引用次数: 0

Deletion of maternal CD163 PSTII-domain-coding exon 13 protects fetuses from infection with porcine reproductive and respiratory syndrome virus (PRRSV) 母体 CD163 PSTII 域编码外显子 13 的缺失可保护胎儿免受猪繁殖与呼吸综合征病毒（PRRSV）的感染

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110255

Raymond R.R. Rowland , Brianna Salgado , James Lowe , Tad S. Sonstegard , Daniel F. Carlson , Kyra Martins , Jonathan R. Bostrom , Suzanna Storms , Alberto Brandariz-Nuñez

Following infection of a porcine dam with PRRSV around 90 days of gestation, the virus crosses the placenta and starts to infect fetuses. This can lead to consequences such as abortions, stillbirths, and respiratory issues in newborn piglets. CD163 is an essential cellular viral entry receptor for porcine reproductive and respiratory syndrome virus (PRRSV). CD163 contains nine scavenger receptor cysteine-rich (SRCR) and two proline-serine-threonine (PST) domains. Gene-edited pigs possessing a complete deletion of CD163 are resistant to PRRSV infection. Recently, we demonstrated that pigs harboring a clean deletion of CD163 exon 13 (ΔExon13 CD163 pigs) which encodes the first 12 amino acids of the CD163 PSTII domain were not susceptible to PRRSV infection. In this study, ΔExon13 CD163 (−/−) gilts were bred with wildtype CD163 (+/+) boars producing heterozygous, CD163 (+/−) fetuses. We found that fetuses with a wildtype CD163, recovered between day 103 of gestation or 17 days after the maternal infection with PRRSV, were fully protected from PRRSV in dams containing a clean deletion of CD163 exon 13. These findings suggest a feasible approach for eliminating PRRSV-related reproductive illness, which is a significant cause of economic losses in agriculture.

猪母体在妊娠 90 天左右感染 PRRSV 后，病毒穿过胎盘开始感染胎儿。这会导致流产、死胎和新生仔猪呼吸道疾病等后果。CD163 是猪繁殖与呼吸综合征病毒 (PRRSV) 重要的细胞病毒进入受体。CD163 包含九个富含半胱氨酸的清道夫受体 (SRCR) 和两个脯氨酸-丝氨酸-苏氨酸 (PST) 结构域。完全缺失 CD163 的基因编辑猪对 PRRSV 感染具有抵抗力。最近，我们证明了CD163外显子13（编码CD163 PSTII结构域的前12个氨基酸）完全缺失的猪（ΔExon13 CD163猪）对PRRSV感染不敏感。在这项研究中，ΔExon13 CD163（-/-）后备母猪与野生型 CD163（+/+）公猪配种，产生了杂合的 CD163（+/-）胎儿。我们发现，在妊娠第 103 天或母体感染 PRRSV 17 天后恢复的 CD163 野生型胎儿，在含有 CD163 第 13 号外显子纯合缺失的母体中完全不受 PRRSV 的感染。这些发现为消除与 PRRSV 相关的生殖疾病提供了一种可行的方法，这种疾病是造成农业经济损失的一个重要原因。

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引用次数: 0

IL-10 upregulates SOCS3 to inhibit type I interferon signaling to promote PoRVA replication in intestinal epithelial cells IL-10 上调 SOCS3，抑制 I 型干扰素信号，促进肠上皮细胞中 PoRVA 的复制

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110259

Haixin Liu , Yongpan Zhao , Huimin Du , Pengcheng Hao , Haolun Tian , Kun Wang , Yudong Qiu , Haiying Dong , Qian Du , Dewen Tong , Yong Huang

Porcine group A rotavirus (PoRVA) is one of the common enteric viruses causing severe diarrhea in piglets. Although PoRVA infection has been identified to promote IL-10 production, the role of IL-10 during viral infection remains unclear. In this study, we found that elevated IL-10 levels during PoRVA infection promote viral replication by inhibiting type I interferon production and response. IL-10 treatment upregulated the expression of SOCS3 in PoRVA-infected IPEC-J2 cells, which inhibited IFN-I production by preventing the degradation of IκB and nuclear translocation of NF-κB, thereby significantly promoting PoRVA replication. Furthermore, we determined that SOCS3 also inhibited type Ⅰ interferon signaling pathway, which led to a significantly reduced ISGs after IFN-α stimulation. In PoRVA-infected cells, overexpression of SOCS3 significantly inhibits phosphorylation and heterodimerization of STAT1, thereby promoting viral replication. Finally, we demonstrated the effect of IL-10 on PoRVA replication in vivo by murine models of PoRVA infection. PoRVA replication levels were lower in the ileum of IL-10 knockout (IL-10^-/-) mice than that in PoRVA-infected wild-type mice, but PoRVA replication levels were higher in the ileum of IFNAR knockout (IFNAR^-/-) mice than that in PoRVA-infected wild-type mice. Taken together, our findings provide information to understand the strategies of PoRVA to evade host innate antiviral immunity.

猪 A 组轮状病毒（PoRVA）是导致仔猪严重腹泻的常见肠道病毒之一。虽然已发现 PoRVA 感染会促进 IL-10 的产生，但 IL-10 在病毒感染过程中的作用仍不清楚。在这项研究中，我们发现 PoRVA 感染期间 IL-10 水平升高会抑制 I 型干扰素的产生和反应，从而促进病毒复制。IL-10处理可上调SOCS3在感染PoRVA的IPEC-J2细胞中的表达，SOCS3通过阻止IκB降解和NF-κB核转位来抑制IFN-I的产生，从而显著促进PoRVA的复制。此外，我们还发现 SOCS3 还能抑制Ⅰ型干扰素信号通路，从而使 IFN-α 刺激后的 ISGs 显著减少。在 PoRVA 感染的细胞中，过表达 SOCS3 能明显抑制 STAT1 的磷酸化和异二聚化，从而促进病毒复制。最后，我们通过小鼠 PoRVA 感染模型证明了 IL-10 对 PoRVA 在体内复制的影响。IL-10基因敲除（IL-10-/-）小鼠回肠中的PoRVA复制水平低于感染PoRVA的野生型小鼠，但IFNAR基因敲除（IFNAR-/-）小鼠回肠中的PoRVA复制水平高于感染PoRVA的野生型小鼠。综上所述，我们的研究结果为了解 PoRVA 逃避宿主先天性抗病毒免疫的策略提供了信息。

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引用次数: 0

Z-Ligustilide restricts rabies virus replication by inducing ferroptosis through the ACSL4-LPCAT3-POR pathway Z-女贞苷通过 ACSL4-LPCAT3-POR 途径诱导铁变态反应，从而限制狂犬病病毒的复制

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110260

Jianqing Zhao , Qianruo Wang , Zhenkun Liu , Meixin Sun , Rui Zhou , Zhen F. Fu , Ling Zhao , Ming Zhou

Rabies, induced by rabies virus (RABV), still threaten global health all over the world, and no effective therapy is available for rabies currently. Recently, a series of natural plant components have been found to inhibit virus production. In this study, Z-Ligustilide, a natural component of Ligusticum chuanxiong Hort, was found to inhibit RABV replication. Initially, the concentration of cytotoxicity 50 % (CC₅₀) of Z-Ligustilide in N2a and BSR cells were 429.9 μM and 335.5 μM, respectively, which both significantly restrict RABV production in a concentration-dependent manner. Moreover, Z-Ligustilide was found to mainly inhibit the replication stage of RABV. Specifically, Z-Ligustilide can suppress lipid droplet (LD) formation via directly inhibiting diacylglycerol acyltransferase 1/2 (DGAT1/2) expression, which can further promote cellular lipid peroxidation, Fe²⁺ concentration, reactive oxygen species (ROS), and induce ferroptosis ultimately. Furthermore, Z-Ligustilide was demonstrated to increase ferroptosis via Acyl-CoA synthetase long-chain family member 4 (ACSL4)- Lysophosphatidylcholine Acyltransferase 3 (LPCAT3)- Cytochrome P450 Oxidoreductase (POR) pathway. Above all, this study explored the antiviral function of Z-Ligustilide, which provides a novel insight for developing anti-RABV drugs.

由狂犬病毒（RABV）诱发的狂犬病仍然威胁着全世界的健康，目前尚无有效的狂犬病治疗方法。最近，人们发现一系列天然植物成分可以抑制病毒的产生。本研究发现，川芎中的天然成分 Z-藁本内酯能抑制 RABV 的复制。最初，Z-女贞苷在 N2a 细胞和 BSR 细胞中的细胞毒性 50% 浓度（CC50）分别为 429.9 μM 和 335.5 μM，两者都以浓度依赖的方式显著限制了 RABV 的产生。此外，研究还发现 Z-女贞苷主要抑制 RABV 的复制阶段。具体来说，Z-女贞苷可通过直接抑制二酰甘油酰基转移酶 1/2（DGAT1/2）的表达来抑制脂滴（LD）的形成，从而进一步促进细胞脂质过氧化、Fe2+浓度、活性氧（ROS），并最终诱导铁变态反应。此外，Z-女贞苷还能通过酰基-CoA合成酶长链家族成员4（ACSL4）-溶血磷脂酰胆碱酰基转移酶3（LPCAT3）-细胞色素P450氧化还原酶（POR）途径增加铁变态反应。总之，这项研究探索了 Z-女贞苷的抗病毒功能，为开发抗 RABV 药物提供了新的视角。

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引用次数: 0

Optineurin inhibits IBDV replication via interacting with VP1 Optineurin 通过与 VP1 相互作用抑制 IBDV 复制

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-23 DOI: 10.1016/j.vetmic.2024.110261

Zhixuan Xiong , Qinghua Zeng , Ying Hu , Chongde Lai , Huansheng Wu

Avibirnavirus, specifically Infectious Bursal Disease Virus (IBDV), is a highly contagious pathogen that causes significant economic losses in the poultry industry. The polymerase protein VP1 of IBDV is critical to the viral life cycle, facilitating the synthesis of viral mRNA and the genome. Previous studies have suggested that various host factors influence the regulation of IBDV polymerase activity. In this study, we identified that IBDV infection induces the expression of optineurin (OPTN), a mitophagy receptor and a protein associated with amyotrophic lateral sclerosis (ALS), as well as a negative regulator of interferon I production. The induced expression of OPTN acts as a suppressor of IBDV replication, a function dependent on its ubiquitin-binding domain (UBAN). Furthermore, we demonstrated that OPTN exerts its antiviral effects through direct interactions with VP1 and VP3, which inhibit the polymerase activity of VP1 by preventing K63-linked ubiquitination of VP1. To our knowledge, this study is the first to report that OPTN, upregulated during IBDV infection, functions as a novel antiviral host factor that limits the virus's replicative capacity, offering a potential target for anti-IBDV therapeutic strategies.

阿维巴尼病毒，特别是传染性法氏囊病病毒（IBDV），是一种传染性极强的病原体，给家禽业造成了巨大的经济损失。IBDV 的聚合酶蛋白 VP1 对病毒的生命周期至关重要，可促进病毒 mRNA 和基因组的合成。以前的研究表明，各种宿主因素会影响 IBDV 聚合酶活性的调节。在这项研究中，我们发现 IBDV 感染会诱导 optineurin（OPTN）的表达，OPTN 是一种有丝分裂受体，是一种与肌萎缩性脊髓侧索硬化症（ALS）相关的蛋白质，也是干扰素 I 生成的负调控因子。OPTN 的诱导表达可抑制 IBDV 复制，这一功能依赖于其泛素结合结构域（UBAN）。此外，我们还证明了 OPTN 是通过与 VP1 和 VP3 直接相互作用来发挥其抗病毒作用的，后者通过阻止 VP1 的 K63 链接泛素化来抑制 VP1 的聚合酶活性。据我们所知，本研究首次报道了在 IBDV 感染过程中上调的 OPTN 作为一种新型抗病毒宿主因子限制了病毒的复制能力，为抗 IBDV 治疗策略提供了一个潜在靶点。

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引用次数: 0

Response of Mycobacterium avium subsp. paratuberculosis isolates to reactive oxygen stress generated by treatment with copper ions 副结核分枝杆菌分离株对铜离子处理产生的活性氧压力的反应。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-21 DOI: 10.1016/j.vetmic.2024.110251

P. Steuer , H.W. Barkema , C. Tejeda , J.M. Hernández , F. Ulloa , M. Salgado

Copper (Cu) ions have been recognized for their efficacy in inactivating bacteria, including Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne’s disease (JD) known for its resilience to unfavorable conditions. However, the response of MAP isolates isolated from cows to Cu exposure remains inadequately understood, as their responses may differ from those of laboratory-adapted reference strains. In this study, we examined the response of MAP isolates obtained from MAP-infected and affected cows to Cu ion treatment, comparing that with the response of the reference strain ATCC 19698 to the same treatment. Three MAP field isolates and the MAP reference strain were exposed to Cu ions, and their viability, protein/lipid damage, ROS production, and gene expression were evaluated in triplicate. Survival differed among isolates, with an isolate from a cow with clinical JD exhibiting increased tolerance to Cu exposure. While Cu treatment induced lipid peroxidation and ROS production across all isolates, genes associated with Cu detoxification and virulence were upregulated, particularly in the reference strain. Whole genome sequencing analysis revealed that, despite genomic similarities between field isolates and the reference strain ATCC 19698, there were differences regarding the presence/absence of genes related with certain virulence factors. Further research on Cu exposure with larger numbers of MAP isolates is needed to explain the stress-induced responses that influence MAP survival during natural infections and in challenging environments.

铜（Cu）离子在灭活细菌（包括副结核分枝杆菌（MAP））方面的功效已得到公认，而副结核分枝杆菌是约翰氏病（JD）的致病菌，以其对不利条件的适应能力而闻名。然而，从奶牛体内分离出的 MAP 菌株对铜暴露的反应仍未得到充分了解，因为它们的反应可能与实验室适应性参考菌株的反应不同。在本研究中，我们研究了从MAP感染和患病奶牛体内分离出的MAP菌株对铜离子处理的反应，并将其与参考菌株ATCC 19698对相同处理的反应进行了比较。将三种 MAP 现场分离株和 MAP 参考菌株暴露于 Cu 离子，并以一式三份的方式评估它们的存活率、蛋白质/脂质损伤、ROS 生成和基因表达。不同分离株的存活率不同，其中来自临床 JD 牛的分离株对铜暴露的耐受性更强。虽然铜处理会诱导所有分离株发生脂质过氧化反应和产生 ROS，但与铜解毒和毒力相关的基因上调，尤其是在参考菌株中。全基因组测序分析表明，尽管野外分离菌株与参考菌株 ATCC 19698 的基因组相似，但在某些毒力因子相关基因的存在/缺失方面存在差异。需要对更多的 MAP 分离物进行铜暴露的进一步研究，以解释影响 MAP 在自然感染和挑战性环境中存活的压力诱导反应。

{"title":"Response of Mycobacterium avium subsp. paratuberculosis isolates to reactive oxygen stress generated by treatment with copper ions","authors":"P. Steuer , H.W. Barkema , C. Tejeda , J.M. Hernández , F. Ulloa , M. Salgado","doi":"10.1016/j.vetmic.2024.110251","DOIUrl":"10.1016/j.vetmic.2024.110251","url":null,"abstract":"<div><div>Copper (Cu) ions have been recognized for their efficacy in inactivating bacteria, including <em>Mycobacterium avium</em> subsp. <em>paratuberculosis</em> (MAP), the causative agent of Johne’s disease (JD) known for its resilience to unfavorable conditions. However, the response of MAP isolates isolated from cows to Cu exposure remains inadequately understood, as their responses may differ from those of laboratory-adapted reference strains. In this study, we examined the response of MAP isolates obtained from MAP-infected and affected cows to Cu ion treatment, comparing that with the response of the reference strain ATCC 19698 to the same treatment. Three MAP field isolates and the MAP reference strain were exposed to Cu ions, and their viability, protein/lipid damage, ROS production, and gene expression were evaluated in triplicate. Survival differed among isolates, with an isolate from a cow with clinical JD exhibiting increased tolerance to Cu exposure. While Cu treatment induced lipid peroxidation and ROS production across all isolates, genes associated with Cu detoxification and virulence were upregulated, particularly in the reference strain. Whole genome sequencing analysis revealed that, despite genomic similarities between field isolates and the reference strain ATCC 19698, there were differences regarding the presence/absence of genes related with certain virulence factors. Further research on Cu exposure with larger numbers of MAP isolates is needed to explain the stress-induced responses that influence MAP survival during natural infections and in challenging environments.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"298 ","pages":"Article 110251"},"PeriodicalIF":2.4,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142376109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Lumpy skin disease virus enters into host cells via dynamin-mediated endocytosis and macropinocytosis 瘤状皮肤病病毒通过达纳明介导的内吞和大蛋白细胞吞噬作用进入宿主细胞。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-19 DOI: 10.1016/j.vetmic.2024.110254

Shasha Wang , Pengyuan Cheng , Ke Guo , Shanhui Ren , Berihun Afera Tadele , Zhengji Liang , Yuefeng Sun , Xiangping Yin , Xiangwei Wang

Lumpy skin disease virus (LSDV), a ruminant poxvirus of the Capripoxvirus genus, is the etiologic agent of an economically important cattle disease categorized as a notifiable disease by the World Organization for Animal Health. However, the endocytic pathway and their regulatory molecules have not been characterized for LSDV. In the present study, specific pharmacological inhibitors were used to analyze the mechanism of LSDV entry into Mardin-Darby Bovine Kidney cell (MDBK) and bovine mammary epithelial cell (BMEC). The results showed that LSDV entered MDBK and BMEC cells depends on low-pH conditions and dynamin. However, the inhibitor of caveolae- and clathrin-mediated endocytosis cann’t inhibit LSDV entry into MDBK and BMEC cells. Furthermore, treatment with specific inhibitors demonstrated that LSDV entry into MDBK and BMEC cells via macropinocytosis depended on the Na1/H1 exchanger (NHE) but not phosphatidylinositol 3-kinase (PI3K). In addition, results demonstrated that these inhibitors inhibited LSDV entry but did not have effect on LSDV binding. Taken together, our study demonstrated that LSDV enters MDBK and BMEC cells through macropinocytosis pathway in a low-PH- and dynamin-dependent manner while independent on PI3K. Results presented in this study potentially provides insight into the entry mechanisms of LSDV, and it may facilitate the development of therapeutic interventions.

结节性皮肤病病毒（LSDV）是一种反刍动物痘病毒，属于 Capripoxvirus 属，是一种具有重要经济价值的牛病的病原体，被世界动物卫生组织列为应通报的疾病。然而，LSDV 的内细胞途径及其调控分子尚未定性。本研究利用特异性药理抑制剂分析了 LSDV 进入马尔丁-达比牛肾细胞（MDBK）和牛乳腺上皮细胞（BMEC）的机制。结果表明，LSDV进入MDBK和BMEC细胞依赖于低pH条件和dynamin。然而，洞穴内和凝集素介导的内吞抑制剂不能抑制 LSDV 进入 MDBK 和 BMEC 细胞。此外，用特异性抑制剂处理后发现，LSDV 通过大蛋白胞吞作用进入 MDBK 和 BMEC 细胞依赖于 Na1/H1 交换子（NHE），而不是磷脂酰肌醇 3- 激酶（PI3K）。此外，研究结果表明，这些抑制剂抑制了 LSDV 的进入，但对 LSDV 的结合没有影响。综上所述，我们的研究表明，LSDV 是以低 PH 和达达敏依赖的方式通过巨细胞吞噬途径进入 MDBK 和 BMEC 细胞的，而不依赖于 PI3K。本研究的结果可能有助于深入了解 LSDV 的进入机制，并有助于开发治疗干预措施。

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引用次数: 0