致病性嗜水气单胞菌及其所致鱼病综述

尽管早在1891年就有因嗜水气单胞菌(Aeromonas hydrophila)感染引致蛙“红腿病”的报道,但由于该菌在自然界尤其是水中广泛分布,一般为正常共栖菌,因此长期以来对其致病性并未重视。近年来在我国南方各省淡水养殖鱼类流行暴发性传染病,有报道系为该菌所致败血症。此外在其它水生经济动物、哺乳动物亦有类似发现。人类因之而发生胃肠炎及伤口感染的病例也日渐增多。嗜水气     

Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea

The prevalence of two serotypes of Streptococcus parauberis isolated from the olive flounder, Paralichthys olivaceus, was evaluated in a total of 29 isolates between 2003 and 2010 in Korea. Streptococcus parauberis isolates were divided into two serologically distinct types (serotype 1 and serotype 2), except for one strain (S1091), using an agglutination assay with rabbit antiserum, and serotype 1 was identified as the dominant type (24 of 29 isolates) in this study. To identify the characteristics of the two serotypes of S. parauberis, we conducted a biochemical test using the API 20 Strep kit, a transmission electron microscopy (TEM) assay, sequence analysis of 16S‐23S rRNA intergenic spacer region (ISR) and a pathogenicity test. In TEM, both serotypes possessed polysaccharide capsule layers around the cell surface when bacterial cells were treated with a homologous serotype of rabbit antiserum. However, we were unable to discriminate serotype‐specific biochemical characteristics and genetic characteristics of 16S‐23S rRNA ISR between the two serotypes. In the pathogenicity test, the serotype 1 strains induced significantly higher mortality than the serotype 2 strains in olive flounder when experimentally inoculated via the intraperitoneal route.     

感染茶假眼小绿叶蝉的金龟子绿僵菌菌株筛选

采用喷雾法测定8株金龟子绿僵菌菌株对假眼小绿叶蝉的致病力。结果表明:以浓度(1±0.5)×107孢子·mL-1的绿僵菌孢子悬液接种12 d后,各菌株对假眼小绿叶蝉均有一定的致病力,校正死亡率和僵虫率分别为(43.08±0.76)%-(76.92±1.72)%和(40.67±1.25)%-(76.67±1.55)%,LT50为6.76-11.48 d。其中,Ma1775菌株对假眼小绿叶蝉致病力最强,校正死亡率和僵虫率分别为(76.92±1.72)%和(76.67±1.55)%,LT50为6.76 d,6 d的LC50为4.156 3×107孢子·mL-1。通过时间—剂量—死亡率模型参数估算,Ma1775菌株对该虫致死效应较强的时间段为3-5 d,该菌株在假眼小绿叶蝉的生物防治中有很大的应用潜力。     

A new bacterial leaf blight disease of oat (Avena sativa) caused by Pantoea agglomerans in China

A new leaf blight disease of oat (Avena sativa) was observed in many oat fields in Huan county, Gansu Province of China, during 2018–2019. Typical symptoms appeared as yellow necrotic and water-soaked lesions. The lesions developed from tip to base of leaves and eventually resulted in leaf withering. Disease incidence on plants (leaves) was approximately 36%–100%. A gram-negative bacterium was isolated from the necrotic lesions of all samples. Coupled with multigene sequence (16S rRNA and gyrB gene) analyses, Biolog Gen III MicroStation, morphological, physiological, and biochemical characterization identified the pathogen as Pantoea agglomerans. Pathogenicity tests by wounding and injection inoculations in the greenhouse established that P. agglomerans could induce typical symptoms as observed in the field. The infection rate in leaves was 44%–71% after 14 days. In addition, host range tests showed that P. agglomerans could infect other plant hosts, including Sorghum sudanense, Medicago sativa, Fagopyrum esculentum, Setaria italica, and Zea mays. This is the first report of P. agglomerans causing bacterial leaf blight disease (LBD) on oat in China. The current study can provide a foundation for the prevention of this disease in the future.     

昆虫病原线虫致病机制研究进展

昆虫病原线虫与体内细菌共生,专性寄生在昆虫体内,在害虫生物防治中发挥着重要作用。昆虫病原线虫致病性研究是其高效应用的基础,涉及线虫及其共生细菌对昆虫的交互作用。本文针对斯氏属和异小杆属线虫的致病因子及这类线虫与寄主昆虫互作机制进行了综述,为昆虫病原线虫的致病机制研究和应用提供参考。     

草地贪夜蛾病原微孢子虫的鉴定及其致病力分析

为明确从田间采集草地贪夜蛾 Spodoptera frugiperda幼虫体内发现的一种微孢子虫的分类地位和致病性,利用传统形态学观察和分子生物学技术对该微孢子虫进行鉴定,同时采用室内生物活性测定法对其致病性进行分析。结果显示,该微孢子虫的形态学特征与家蚕微孢子虫 Nosemabombycis相近,具有典型微孢子虫超微组成结构,孢子壁厚度为195.00~205.15 nm,极丝盘旋于孢子后极内侧10~12圈;其基因组的基因间隔区（intergenic spacer, ITS）和小亚基核糖体RNA（smallsubunit ribosomal RNA, SSU）序列与已报道的家蚕微孢子虫相关序列的相似度分别达94.34%和99.50%,系统发育树显示该微孢子虫属于微孢子虫属 Nosema,与家蚕微孢子虫亲缘关系最近。该微孢子虫侵染草地贪夜蛾1龄和2龄幼虫5 d时的LC50分别为2.51×10⁷孢子/mL和2.48×10⁷孢子/mL;侵染3龄幼虫10 d时的LC₅₀为3.79×10⁷孢子/mL;侵染4龄幼虫15 d时的LC50为3.98×10⁷孢子/mL;且当微孢子虫浓度为1.0×10⁸孢子/mL时,草地贪夜蛾1至4龄幼虫的LT₅₀分别为3.04、 3.86、 7.47和10.43 d。表明该微孢子虫隶属微孢子虫属,对草地贪夜蛾不同龄期幼虫均有较强的致病力,具有良好的开发应用潜力。     

三株球孢白僵菌的分离鉴定与其对粘虫的室内毒力

通过黄粉虫虫饵法从河北省采集的土样中分离得到三株昆虫病原真菌,结合形态学特征和rDNA序列分析对菌株进行了鉴定,结果表明：三株均为球孢白僵菌Beauveria bassiana,编号依次为Z-G2-5、M-G4、M-G3。通过生测发现3株球孢白僵菌菌株均能有效感染粘虫幼虫,三株球孢白僵菌对粘虫3龄幼虫的毒力不同：在5×10⁸孢子/mL剂量下,菌株Z-G2-5第6 d时对粘虫的累计校正死亡率达到96.77%,LT₅₀为3.02 d,LC₅₀为4.95×10⁶个孢子/mL。而菌株M-G3、M-G4的累计死亡率分别为66.67%、71.11%;LT₅₀分别为4.89 d、3.97 d;LC₅₀分别为1.18×10⁸个孢子/mL、1.13×10⁷个孢子/mL。上述结果表明,相比其他两株菌,菌株Z-G2-5对粘虫的致病力更强,致死速度更快,具有较大的应用价值,可作为粘虫幼虫生物防治的候选菌株。     

云南省草果主要病原真菌鉴定及防治药剂筛选

为了明确引起云南草果植株主要病害的病原真菌种类,筛选能有效抑制分离病原菌的杀菌剂,为草果病害化学防治提供依据。采用组织分离法纯化培养,并通过形态学鉴定并结合ITS,TEF,ACT和GAPDH序列分析确定病原菌的分类地位;通过菌丝生长速率法,明确7种杀菌剂对草果主要病害的防治效果。结果表明,分离到的菌株为禾谷镰刀菌(Fusarium graminearum)和胶孢炭疽菌(Colletotrichum gloeosporioides)。防治药剂筛选结果显示,对于禾谷镰刀菌,所选的7种杀菌剂中京彩的防治效果达到93%以上。对于胶孢炭疽菌,代森锰锌在田间推荐量区间内可以实现百分百防治。蓝楷、京彩、施灰乐低浓度时对胶孢炭疽菌的防治效果均达到了90%以上。推荐京彩、施灰乐,代森锰锌和蓝楷作为云南省草果主要真菌病害的优选兼治药剂。     

球孢白僵菌对假眼小绿叶蝉成虫高致病力菌株筛选

采用喷雾法测定5株球孢白僵菌菌株对假眼小绿叶蝉的致病力。结果表明:以浓度(1±0.5)×107孢子/m L白僵菌孢子悬液接种12 d后,各菌株对假眼小绿叶蝉均有一定的致病性,校正死亡率和僵虫率范围分别为(52.21±0.94)%~(86.92±1.96)%和(50.67±1.25)%~(84.67±0.82)%,LT50为6.35~9.61 d。其中,BLK菌株对其致病力最强,校正死亡率和僵虫率分别为(86.92±1.96)%和(84.67±0.82)%,LT50为6.35 d,6 d的LC50为9.539×106孢子/m L。通过时间-剂量-死亡率模型参数估算,BLK菌株对该虫致死效应较强的时间段为2.5~5 d,该菌株在假眼小绿叶蝉的生物防治中有重要的应用潜力。     

广东省南美白对虾豚鼠气单胞菌的分离鉴定

广东省某养殖场南美白对虾于2012年突发传染病,为确定该病病原并对其进行有效防制,特利用分子生物学技术对病原进行了分离鉴定。结果从发病南美白对虾分离纯化到1株优势菌ZHBX12016,将该菌人工浸浴感染南美白对虾,5 d后南美白对虾死亡率达95%,且发病对虾临床症状与自然病例相似,人工注射感染测得其半数致死量LD50为6.43×103cfu/mL。该菌革兰氏染色阴性,短杆状;对半乳糖、蔗糖、甘露醇、葡萄糖产气、精氨酸双水解酶,赖氨酸脱酸酶,葡萄糖酸盐的生化反应性与豚鼠气单胞菌(Aeromonas caviae)(synonym Aeromonas punctata)生化反应结果相同。用聚合酶链式反应(PCR)技术扩增该菌16S rD NA序列进行测序和生物信息学分析,结果显示:分离菌株与豚鼠气单胞菌(Aeromonas caviae)相似性达97.7%~100%之间,在系统发育树上聚为同一分支。综合该菌形态学、致病性、生理生化和16S rRNA基因分析结果,将其鉴定为豚鼠气单胞菌(Aeromonas caviae)。药敏试验结果表明分离菌株对氯霉素、土霉素、丁胺卡那、呋喃唑酮等高度敏感。