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1. 植物乳杆菌对藤茶副产物青贮品质影响的研究
WANG Yuan, ZHOU Hong-zhang, GAO Yu, WANG Ning-wei, LIU Han, YANG Fu-yu, NI Kui-kui
Journal of Integrative Agriculture    2023, 22 (4): 1172-1183.   DOI: 10.1016/j.jia.2022.10.001
摘要221)      PDF    收藏

本文研究目是探究青贮技术以及植物乳杆菌在藤茶副产物贮藏中的应用实验分为两组,包括对照组和植物乳杆菌处理组。在室温(23-30°C)条件下贮藏7、14和30,分析藤茶副产物青贮发酵品质、营养成分、抗氧化活性以及细菌群落。结果显示,与对照组相比,植物乳杆菌组具有较低(P<0.05)的pH和氨态氮,以及较高(P<0.05)的乳酸含量。虽然总酚在青贮过程中变化较小,但乳酸菌处理组具有较高P<0.05ABTS自由基清除能力。青贮30天后,Firmicutes取代Proteobacteria成为优势菌门,Lactobacillus成为优势属Spearman相关性结果显示乳酸与乳杆菌呈正相关关系(P<0.01)。总之,用植物乳杆菌来青贮藤渣副产物可以有效地改善青贮发酵品质和减少营养物质损失,为藤渣副产物饲料化利用提供了新的研究思路。

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2. JIA-2021-2061 CovS在马链球菌兽疫亚种毒力形成中的作用研究
XU Bin, MA Zhe, ZHOU Hong, LIN Hui-xing, FAN Hong-jie
Journal of Integrative Agriculture    2023, 22 (2): 568-584.   DOI: 10.1016/j.jia.2022.08.109
摘要314)      PDF    收藏

目的:马链球菌兽疫亚种(Streptococcus equi subsp. zooepidemicus,SEZ)是一种人畜共患病病原,在我国主要引起猪链球菌病。本实验室前期研究发现了一株源于强毒株SEZ ATCC35246自然变异的弱毒株M35246M35246表现为一个连续25基因的丢失和covS基因的功能丧失性突变。这是第一次发现在SEZ中的涉及covS的自然变异。涉及covS的自然变异是增强化脓链球菌致病性的关键,所以需要确定covS的自然变异是否对SEZ毒力具有相同的影响。本工作的目的是研究CovSSEZ毒力形成中的作用有助于研究SEZ的致病机制,特别是涉及SEZ毒力的转录调控机制。

方法:本研究通过转录组测序和DNA测序,确定了M35246covS的碱基突变形式。在野生强毒株ATCC35246的基础上分别构建了25基因敲除株ΔPIcovS突变株McovS及对应的互补株。随后,本研究检测了ATCC35246M35246M35246 CcovSMcovSCMcovSΔPI的生长能力、对上皮细胞HEp-2黏附能力、对巨噬细胞Raw264.7的抗吞噬能力以及菌体荚膜含量;测定了ATCC35246M35246McovSΔPI24种抗生素的敏感性、对小鼠的半数致死量和攻毒后的菌体脏器分布;进行了ATCC35246M35246McovS的比较转录组学分析。

结果:M35246covS的变异导致其移码突变并造成提前翻译终止,且在基因N端形成终止子结构,遏制其转录。与ATCC35246相比,M35246McovS的荚膜含量和抗吞噬能力显著降低。McovSβ-内酰胺类、氨基糖苷类、大环内酯类和林可酰胺类药物的敏感性显著高于ATCC35246。与ATCC35246相比,M35246McovSΔPI小鼠的半数致死量分别增加了1051055倍。2000ATCC35246半数致死量的剂量攻毒48小时后,M35246McovS均不能从小鼠体内分离。转录组分析表明,McovSATCC35246之间存在668个显著差异表达的基因。相对于ATCC35246McovS中与抗吞噬、荚膜形成、致病性和抗生素抗性有关的许多毒力因子编码基因和合成代谢相关基因显著下调。

结论:本文系统研究了SEZ CovS在细菌抗吞噬作用、荚膜形成、致病性、抗生素耐药性以及各种重要毒力因子和关键代谢系统转录调控的作用。此外,转录组分析揭示了CovS在抗吞噬作用、荚膜形成、致病性和抗生素耐药性方面的调节机制。

创新性:该工作系统研究了参与SEZ致病性和抗生素耐药的调控因子,表明二元调控系统在不同细菌中调控的多样性,揭示CovSSEZ毒力形成中起着至关重要的作用。

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3. Biology, invasion and management of the agricultural invader: Fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)
Jing WAN, HUANG Cong, LI Chang-you, ZHOU Hong-xu, REN Yong-lin, LI Zai-yuan, XING Long-sheng, ZHANG Bin, QIAO Xi, LIU Bo, LIU Cong-hui, XI Yu, LIU Wan-xue, WANG Wen-kai, QIAN Wan-qiang, Simon MCKIRDY, WAN Fang-hao
Journal of Integrative Agriculture    2021, 20 (3): 646-663.   DOI: 10.1016/S2095-3119(20)63367-6
摘要181)      PDF    收藏

重大农业入侵害虫草地贪夜蛾原产于美洲,自2016年首次被发现入侵尼日利亚和加纳以来,短短3年时间内迅速入侵至47个非洲国家和18个亚洲国家。由于该虫寄主范围广(至少包含353种寄主植物)、能够适应多种生境、超强的迁飞能力、高繁殖力、暴食性,以及快速发展的农药抗性和病毒抗性等内在优势,是导致其具有入侵性的重要原因,目前已被公认为全球范围内的超级害虫。该害虫的综合治理策略主要依靠监测调查、农业防治、化学防治、病毒制剂、性诱剂、生物防治(寄生性天敌、捕食性天敌和昆虫病原体),以及植物源农药等多种防治策略的综合应用。目前尚需要进一步研究的主要内容包括:(1)明确草地贪夜蛾的入侵机制,(2)如何阻止其进一步扩散,(3)提供更有效的防治策略。本文总结了草地贪夜蛾的生物学特性,潜在的入侵性机制,以及综合治理策略,以期为今后的治理提供参考。


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4. Evaluating the efficacy of an attenuated Streptococcus equi ssp. zooepidemicus vaccine produced by multi-gene deletion in pathogenicity island SeseCisland_4
MA Fang, WANG Guang-yu, ZHOU Hong, MA Zhe, LIN Hui-xing, FAN Hong-jie
Journal of Integrative Agriculture    2019, 18 (5): 1093-1102.   DOI: 10.1016/S2095-3119(18)62133-1
摘要147)      PDF    收藏
Streptococcus equi ssp. zooepidemicus (SEZ) is a pathogen associated with a wild range of animal species.  Frequent outbreaks have occurred in recent years in pigs, horses, goats and dogs which is liable to infect humans.  There is a lack of efficient vaccines against this disease and the occurrence of antibiotic resistance may render drug therapies ineffective.  In this study, gene deletion mutant (ΔSEZ) in pathogenicity islands SeseCisland_4 was constructed.  The mutant ΔSEZ had a 52-fold decrease in 50% lethal dose (LD50) and had less capacity to adhere epithelial cells.  Importantly, immunization of mice with attenuated vaccine ΔSEZ at the dose of 102 colony-forming units (CFU) mL–1 elicited a significant humoral antibody response, with an antibody titer of 1:12 800.  Therefore, 102 CFU mL–1 might be used as the appropriate immune dose for the attenuated vaccine ΔSEZ, which provided mice with efficient protection against virulent SEZ.  In addition, the hyperimmune sera against 102 CFU mL–1 attenuated vaccine ΔSEZ could confer significant protection against virulent SEZ infection in the passive immunization experiment and exhibited efficient bactericidal activity in the whole blood assay.  Meanwhile, no viable bacteria was detected in blood when mice were immunized with ΔSEZ at the dose of 102 CFU mL–1 via hypodermic injection.  Thereafter, the mutant ΔSEZ at the dose of 102 CFU mL–1 could confer significant protection in mice and had less negative effects on host, which could be an effective attenuated vaccine candidate for the prevention of SEZ.
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5. Identification and Characterization of Putative Virulent Genes in Streptococcus equi ssp. zooepidemicus
ZHOU Hong, MA Zhe, YUAN Jin , FAN Hong-jie
Journal of Integrative Agriculture    2013, 12 (2): 327-333.   DOI: 10.1016/S2095-3119(13)60232-4
摘要1371)      PDF    收藏
Suppression subtractive hybridization (SSH) was performed with virulent strain ATCC35246 and avirulent strain ST171 to identify novel genes associated with virulence in Streptococcus equi ssp. zooepidemicus (SEZ). There were fourteen genomic regions that only presented in virulent strain ATCC35246. These regions encoded 14 proteins, some of them were homologous to proteins associated with cellular surface structure, molecular synthesis, energy metabolism, regulation, transport systems, and other unknown functions. Primers for 6 particular regions were designed from the already published SEZ sequence. Then, we used PCR to evaluate the distribution and conservation of these 6 DNA fragments in various SEZ strains collected from different sources, regions, groups, and times. The results showed that these 6 DNA fragments were widely distributed in SEZ strains, yet they were not existence in the avirulent strain ST171. Moreover, these fragments could not be detected in other Streptococcus groups.
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