A bacterium with close genetic identity to Pseudomonas mandelii associated with spring fish kills in wild bluegill Lepomis macrochirus Rafinesque and pumpkinseed sunfish Lepomis gibbosus (Linnaeus)

Pseudomonas fluorescens are known bacterial pathogens in fish. The P. fluorescens group contains at least nine different bacterial species, although species from fish have rarely been differentiated. Two isolated fish kills affecting wild bluegills, Lepomis macrochirus Rafinesque, and pumpkinseed sunfish, Lepomis gibbosus (Linnaeus), occurred in the spring of 2015 during cool water temperatures (12.5°C–15.5°C). Disease signs included severe bacteraemia with rare gross external signs. Pure bacterial cultures isolated from kidneys of all affected fish were identified as P. fluorescens using the API 20NE system, while no bacteria were isolated from asymptomatic fish. To further identify the species of bacterium within the P. fluorescens complex, genetic analysis of the 16S rRNA, rpoD and gyrB genes was conducted. DNA sequences of bacterial isolates from both mortality events were identical and had close identity (≥99.7%) to Pseudomonas mandelii. Although likely widespread in the aquatic environment, this is the first report of a bacterium closely resembling P. mandelii infecting and causing disease in fish. The bacterium grew at temperatures between 5°C and 30°C, but not at 37°C. It is possible that infections in fish were a result of immunosuppression associated with spring conditions combined with the psychrotrophic nature of the bacterium.     

Response of Bacterial Biofilms in Recirculating Aquaculture Systems to Various Sanitizers

ABSTRACT

Pathogenic microorganisms may be incorporated into biofilms found in aquaculture systems, causing recurring exposure to potential disease agents. Aerobic plate counts, the presence of Enterobacteriaceae, and the presence of Escherichia coli, modified to express a green fluorescent protein (GFP E. coli), was used to evaluate the effectiveness of various sanitizers in decreasing bacterial incorporation into newly generated biofilms in recirculating aquaculture systems. Disks of Buna-N rubber, polyvinyl chloride (PVC), chlorinated PVC, glass, fiberglass, and stainless steel were placed in aquariums stocked with Nile tilapia, Oreochromis niloticus. The effectiveness of water, an alkaline cleanser, sodium hypochlorite, a quaternary ammonium compound, or peracetic acid as a sanitizer was evaluated on each substrate by enumerating total plate counts, GFP E. coli, and Enterobacteriaceae. Sodium hypochlorite and peracetic acid were the most effective sanitizers, with an overall percentage reduction of GFP E. coli of approximately 2 logs₁₀. The quaternary ammonium compound was moderately effective, 1 log₁₀, against the target organisms. Water demonstrated a 2 log₁₀ reduction of the total plate count, suggesting that some mechanical cleaning was achieved. The type of material used as substrate for the biofilm had no significant (P?>?0.05) effect on the effectiveness of the sanitizers.     

氨基酸菌肥施用量对茶树生长及土壤肥力的影响

为探究不同氨基酸菌肥施用量对茶树生长和土壤肥力含量的影响,以鸿雁12号茶树为研究对象,采用田间微区试验的方法,设置了不施用氨基酸菌肥作对照组(CK)和氨基酸菌肥施用量分别为75 kg·hm^-2(T1)、150 kg·hm^-2(T2)、225 kg·hm^-2(T3)和300 kg·hm^-2(T4)的处理,研究不同氨基酸菌肥施用量对茶树生长、茶叶品质及土壤肥力的影响。结果表明：氨基酸菌肥施用后能显著增加茶叶芽头密度及茶叶产量,提高茶叶中氨基酸含量和土壤中碱解氮含量。试验6个月后与CK相比,T4组的芽头密度显著增加了63.4%,茶叶产量显著增加了71.5%;T1～T4处理茶叶中氨基酸含量分别增加了9.73%、6.41%、7.57%和13.2%,土壤中碱解氮含量分别增加了25.0%、36.0%、46.0%和37.6%。综上,氨基酸菌肥的施用能提高茶叶产量及品质,改善茶园土壤环境质量,为茶园生产的合理施肥及优质高效生产提供数据参考。     

纤维素降解菌的筛选、鉴定和糖化水平研究

将前期从木薯生境中分离、保存的298个菌株,通过平板测试,筛选到57个可产纤维素酶活的菌株,其中酶活性效果最好的菌株(水解圈最大,将其赋值得分为3)所占比例为40.9%,酶活性依次减弱的菌株所占比例(赋值得分为2和1)分别占27.9%和24.6%。采用DNS法测定57个菌株的糖化水平和酶活力,结果显示,10株细菌具有较好潜力降解纤维素,其中菌株HWY-3-9的糖化和酶活力最好,产糖量为0.1852 mg/m L,酶活力为0.0012 U/m L,经鉴定为解淀粉芽孢杆菌。     

对暗褐网柄牛肝菌菌丝生长有益细菌的筛选

从用于暗褐网柄牛肝菌(Phlebopus portentosus)人工栽培的覆土和草炭中分离、纯化出40株细菌,经液体发酵试验和平板试验检测这些菌株对暗褐网柄牛肝菌菌丝生长的影响,筛选出对暗褐网柄牛肝菌菌丝生长有明显促进作用的3株细菌(编号为:T24、T34、C1)。     

抑菌圈-定殖力双重测定法筛选青枯病生防细菌

 本研究首先用平皿抑菌圈法筛选出55个拮抗青枯菌的细菌菌株,将番茄幼苗在各菌菌悬液中浸根12h后栽种于温室未灭菌的土壤中,结果发现,有22个菌株在幼苗根部的定殖能力较强(终定殖密度大于104 cfu/g根),其中革兰氏阴性土壤细菌占同类菌的86.3%,革兰氏阳性土壤细菌占同类菌的13.0%,10个无致病力产细菌素的青枯菌菌株在根部的终定殖密度均低于104 cfu/g根,其定殖能力弱于致病青枯菌。有17个拮抗菌菌株在番茄幼苗根部的定殖密度超过所有致病菌。温室生防结果证明,抑菌圈-定殖力双重测定法对于筛选生防菌株是一种行之有效的方法。     

番茄酱中好氧细菌分离方法的研究

新疆番茄制品出口量占全国的90;,近年来外商对其安全卫生提出了许多技术要求,国内番茄制品生产企业也陷入番茄制品胖听、平听的技术障碍,至今国内尚无相应的微生物检测技术和标准.采用锰营养、甘露醇卵黄多粘菌素、嗜热耐酸杆菌、脂耐热嗜酸杆菌、溴甲酚紫葡萄糖肉汤、平酸菌增菌和液体硫乙醇酸盐7种液体培养基对供试番茄酱在37和55℃下培养.通过不同培养时间培养液的O.D值、pH值、菌体形态和菌落特征的研究,确定出锰营养和平酸菌增菌培养基为该类菌的最适液体培养基,嗜热耐酸杆菌和平酸菌增菌琼脂培养基为最佳固体培养基.分离出的菌株经VITEK-32生化鉴定,主要细菌分别为枯草芽孢杆菌(Bacillus subtilis)、地衣芽孢杆菌(Bacillus licheniformis)和矮小芽孢杆菌(Bacillus pumilus),均为产酸不产气可导致平酸腐败的好氧芽孢菌.     

Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens

An isolate of Trichoderma harzianum Rifai from an infected cacao pod produces and secretes nonanoic (pelargonic) acid into a liquid culture medium. Nonanoic acid (NA) was very inhibitory to spore germination and mycelial growth of two cacao pathogens, Crinipellis perniciosa Stahel and Moniliophthora roreri Cif. H.C. Evans. It was highly active causing 75% inhibition of spore germination in an in vitro assay at a rate as low as 0.09 μM for M. roreri and 0.92 μM for C. perniciosa. Mycelial growth was comparatively less sensitive to inhibition, but still there was a 75% reduction in growth with 0.62 μM in M. roreri and 151 μM NA in C. perniciosa. In contrast, NA did not affect Trichoderma mycelial growth or spore germination at concentrations that were inhibitory to the pathogens. 6-pentyl-α-pyrone was also produced and secreted into the medium by T. harzianum, however; it was not antagonistic to the cacao pathogens. Although a number of metabolites produced by Trichoderma spp. have been identified in the past, this is the first report of NA production and secretion by any Trichoderma. The results suggest that NA may play a role in the successful use of some Trichoderma spp. isolates in the biocontrol of fungal diseases of plants.     

冰核细菌的定量分离研究

 冰核细菌可能是自然界中诱发和加重霜冻敏感植物发生霜冻害的重要原因之一。由于单个的冰核细菌细胞并不具有冰核活性,因此,研究植物上冰核细菌的种群数量或密度是研究冰核细菌与植物霜冻害关系的关键。从云南省的元江、寻甸、河口、呈贡、临沧、陆良、宣威、曲靖、富民、昆明市官渡区及越南老街等地采集到108种植物共166个标样中,有45个标样分离共到71株冰核细菌,标样中冰核细菌的检出率为27.1％,最低密度为6.0×10⁴ CFU/g,最高密度为5.12×10⁹ CFU/g。其余121个标样经再次分离未检测冰核细菌,占总标样数的72.9％。在云南植物上,用稀释平板法可检测冰核细菌的最低密度为2.0×10⁴ CFU／g.     

两种载体生物膜中异养细菌数量动态及其氨化作用

用微生物学方法测定了淹没式生物滤器中两种载体生物膜形成过程中异养细菌数量动态及熟化后细菌的氨化作用。结果表明,载体生物膜形成过程中,异养细菌呈逐渐减少的趋势,生物滤器运行前期异养细菌数量高(107～108CFU/g),随着时间的推移逐渐减少(105～106CFU/g);沸石载体生物膜细菌的氨化速率前24h平均为17 45μg/(cm3·h)和26μg/(g·h),无纺布载体生物膜细菌的氨化速率前24h平均为17 41μg/(cm3·h)和59 9μg/(g·h)。