pyrG基因对西瓜噬酸菌致病性和组氨酸利用的影响

为研究pyr G基因与西瓜噬酸菌(Acidovorax citrulli)致病相关性,利用同源重组的方法构建pyr G基因突变体(Δpyr G),对pyr G基因的功能进行分析。结果显示,pyr G基因影响病原菌的致病性、游动性、生物膜形成和定殖能力以及激发烟草过敏反应能力;pyr G基因影响his D基因的表达;此外,pyr G基因突变影响西瓜噬酸菌对组氨酸的利用。     

一株尼罗罗非鱼致病性嗜水气单胞菌的 分离鉴定及药敏试验

从自然感染发病的尼罗罗非鱼肝脏和脑组织中分离出1 株细菌,经革兰氏染色镜检、生化鉴定和PCR 鉴定,确定为嗜水气单胞菌,命名为GD001,进一步采用药敏纸片法测定分离菌株对多种抗生素的敏感性,并用该菌 株接种健康尼罗罗非鱼,鉴定其致病力。结果发现,GD001 菌株对恩诺沙星、头孢噻吩、四环素和克莱霉素等抗生素 敏感,对链霉素轻度敏感,对卡那霉素和青霉素不敏感。从人工感染10 g 左右的健康罗非鱼发现,GD001 菌株具有很 强的致病力,试验鱼在感染后3~4 h内全部死亡,且患病症状与自然发病症状的鱼一致。     

淡紫拟青霉T-DNA插入突变体的表型分析

研究淡紫拟青霉T-DNA插入突变体,鉴定突变体的表型特征,为淡紫拟青霉致病机制分析及致病相关的基因克隆奠定基础。PCR检测筛选的14株T-DNA插入突变体,观察和测定菌落形态,菌落生长速率,产孢量,孢子萌发率和菌丝干质量等生物学性状,并比较突变体致病力。结果表明突变体均含有beta-tubulin基因序列,T-DNA已整合进入野生型菌株20-7的基因组中。通过与野生型菌株对比,发现突变体菌落形态发生了不同程度的改变。菌落生长速率明显增大的菌株占全部菌株的85.71%,只有BN-11菌株的菌落生长速率降低。产孢量显著变大的菌株有BN-11和BN-12,为菌株总数的14.29%。孢子萌发率发生改变的菌株占78.57%。菌丝干质量发生明显增大和减少的菌株都占全部突变体的21.43%。筛选获得了1株致病力增强的突变体BN-11和11株致病力显著降低的突变体。     

一株GPV的分离鉴定及致病性研究

为了确定导致吉林省某地区10日龄雏鹅发病的病原,从病死雏鹅的肝脏中分离到一株病毒,根据病毒的电镜照片,动物回归试验及PCR鉴定,确定为鹅细小病毒。PCR产物测序后经BLAST比对显示,分离毒株与NCBI收录的GPVVP3基因同源性均在92％以上,与GPV／CH／HLJ02／08VP3基因的同源性最高,达99％。致病性研究表明,分离株的ELD50为10^-6.5／0．2mL,毒力较强,且该毒株能被GPV标准血清所中和。     

Molecular characterization and pathogenicity of a grouper iridovirus (GIV) isolated from yellow grouper, Epinephelus awoara (Temminck & Schlegel)

Molecular characterization was carried out on an iridovirus isolated from yellow grouper, Epinephelus awoara . The major capsid protein (MCP) gene was located, sequenced and compared with homologous genes from other iridoviruses. The nucleotide sequence is 1392 bases long and contains a single open reading frame beginning at an ATG codon from the 5' end and terminating at a TAA codon at the 3' end. The open reading frame encodes a protein of 463 amino acids with a predicted molecular weight of 50 272 Da. Pairwise amino acid alignments detected a high degree of sequence identity between grouper iridovirus (GIV) MCP and the homologous genes of other iridoviruses. The MCP gene of GIV was most similar to the MCP gene from frog virus 3 (FV3) with 70% nucleotide and 73% amino acid sequence identity. The predicted molecular weight of the protein of this gene is comparable with the apparent weight obtained by SDS–PAGE. Pathogenicity of the GIV was investigated in yellow grouper by intraperitoneal injection of 10⁷ and 10⁴ TCID₅₀ virus. Cumulative mortalities reached 100% within 11 and 25 days post-infection, respectively, while no grouper died in the control group. The molecular studies demonstrated that GIV is a member of the genus Ranavirus .     

The pathogenic helminth parasites of eels

Although 63 and 55 species of helminths have been reported from each species of Atlantic eel and from 29 to 19 for each species of Pacific eel only the monogeneans Pseudodactylogyrus bini and P. anguillae and the nematode Anguillicola crassus, originally specific to species of Pacific eels, can be considered serious pathogens. None of the three are normally pathogenic to their preferred natural eel host species in the wild. Pseudodactylogyrus spp. only cause serious local gill damage when present on a host in large numbers under optimal conditions that facilitate transmission. This is the case in eel aquaculture, where infections can be controlled by drugs. Anguillicola crassus is only pathogenic to Anguilla anguilla and A. rostrata when Atlantic eels are introduced to the far east or when the parasites have been introduced to Europe. Here the parasite life cycle differs in that A. crassus can infect a wide range of intermediate hosts, employ paratenic hosts and survive as larvae for months in the swimbladder wall. This makes it an excellent colonizer. Its major pathogenic effects on eels result from haemorrhaging in, and thickening of, the swimbladder wall. It reduces the oxygen concentration in the swimbladder, reducing its ability to function as a hydrostatic organ, and increases the stress response of eels. In shallow lakes at warm temperatures this can result in mass mortalities. It is also feared that the parasite affects the ability of eels to migrate to the Sargasso Sea and so contributes to the decline in eel populations. Control by drug treatment is possible in culture, but not in the wild.     

大口黑鲈鲁氏耶尔森氏菌的分离鉴定、主要毒力基因及致病性研究

为探明2021年3月福建某水库养殖大口黑鲈疾病暴发的原因,实验从患病大口黑鲈肝脏、肾脏、脾脏中分离优势菌,通过人工感染实验确定病原菌,并综合16S rDNA基因序列分析、生理生化和质谱特征等技术对该病原菌进行种属鉴定,同时,进行毒力基因检测、药物敏感性实验以及组织病理学观察。结果显示,从病鱼脾脏中分离获得1株优势菌并鉴定为鲁氏耶尔森氏菌;该菌株对大口黑鲈的半致死剂量为3.8×10⁵ CFU/尾;该菌株携带yrP1、yhl A、yhl B等毒力基因,对恩诺沙星、盐酸多西环素、氟甲喹、硫酸新霉素、氟苯尼考等5种药物相对敏感。组织病理学观察发现,鲁氏耶尔森氏菌的感染造成大口黑鲈肝脏、肾脏、脾脏不同程度的损伤,表现为明显的变性、坏死及炎症细胞的浸润等。本研究首次报道了鲁氏耶尔森氏菌对养殖大口黑鲈的致病性,可为养殖大口黑鲈鲁氏耶尔森氏菌病的诊断和药物防治提供参考依据。     

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。