Divergence of a strain of Pseudomonas aeruginosa during an outbreak of ovine mastitis

Bacterial infections causing mastitis in sheep can result in severe economic losses for farmers. A large survey of milk samples from ewes with mastitis in Sardinia, Italy, indicated an increasing prevalence of Pseudomonas aeruginosa infections. It has been shown previously that during chronic, biofilm-associated infections P. aeruginosa populations diversify. We report the phenotypic and genomic characterisation of two clonal P. aeruginosa isolates (PSE305 and PSE306) from a mastitis infection outbreak, representing distinct colony morphology variants. In addition to pigment production, PSE305 and PSE306 differed in phenotypic characteristics including biofilm formation, utilisation of various carbon and nitrogen sources, twitching motility. We found higher levels of expression of genes associated with biofilm formation (pelB) and twitching motility (flgD) in PSE305, compared to the biofilm and twitching-defective PSE306. Comparative genomics analysis revealed single nucleotide polymorphisms (SNPs) and minor insertion/deletion variations between PSE305 and PSE306, including a SNP mutation in the pilP gene of PSE306. By introducing a wild-type pilP gene we were able to partially complement the defective twitching motility of PSE306. There were also three larger regions of difference between the two genomes, indicating genomic instability. Hence, we have demonstrated that P. aeruginosa population divergence can occur during an outbreak of mastitis, leading to significant variations in phenotype and genotype, and resembling the behaviour of P. aeruginosa during chronic biofilm-associated infections.     

Characterization of Mannheimia haemolytica biofilm formation in vitro

Mannheimia haemolytica is the primary bacterial agent in the bovine respiratory disease complex. It is thought that M. haemolytica colonizes the tonsillar crypts of cattle as a commensal and subsequently descends into the lungs to cause disease. Many bacterial species persist in the host as biofilms. There is limited information about the ability of M. haemolytica to form biofilms. The aim of this study was to develop an in vitro model for M. haemolytica biofilm formation. We found that M. haemolytica required at least 36 h to form robust biofilms on plastic in vitro when incubated in RPMI-1640 tissue culture medium at 37 °C, with maximal biofilm formation being evident at 48 h. Biofilm formation was inhibited by adding the monosaccharides d(+) galactose and d(+) mannose to the growth medium. Addition of antibodies to the M. haemolytica surface protein OmpA also reduced biofilm formation. Upon evaluating the macromolecules within the biofilm extracellular polymeric substance we found it contained 9.7 μg/cm² of protein, 0.81 μg/cm² of total carbohydrate, and 0.47 μg/cm² of extracellular DNA. Furthermore, proteinase K treatment significantly decreased biofilms (P < 0.05) while α-amylase and micrococcal nuclease decreased biofilms to a lesser extent. M. haemolytica biofilm cells were more resistant than planktonic cells to the antibiotics florfenicol, gentamicin, and tulathromycin. These results provide evidence that M. haemolytica can form biofilms, which could contribute to its ability to persist as a commensal in the bovine upper respiratory tract.     

以玉米芯为碳源和生物膜载体的反硝化反应器启动性能研究

通过小试试验,研究了以玉米芯为反硝化碳源和生物膜载体的生物反应器在启动阶段对污水中COD及硝酸盐的去除效果。结果表明,在模拟废水COD100 mg/L、NO3--N40 mg/L,pH值6.5～7.5条件下,13 d挂膜后间歇处理后的废水COD可降至30 mg/L,18 d挂膜后废水NO3--N降至2 mg/L以下。     

五倍子和石榴皮对溶藻弧菌及其生物膜的体外抑制作用

[目的]明确五倍子(Rhus chinenis)、石榴皮(Folium sennae)对溶藻弧菌(Vibrio alginolyticus)及其生物膜的体外抑制作用。[方法]采用琼脂扩散法,分别测定五倍子、石榴皮对溶藻弧菌的体外抑菌作用;选用倍比稀释法确定五倍子、石榴皮对溶藻弧菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC);采用改良微孔板法评价2种中草药液对溶藻弧菌生物膜形成的影响。[结果]2种中草药都有不同程度的抑制溶藻弧菌的作用,五倍子对溶藻弧菌的抑菌圈直径为(16.37±0.14)mm,MIC和MBC均为6.25 mg/m L;石榴皮对溶藻弧菌的抑菌圈直径为(12.37±0.06)mm,MIC和MBC分别为12.50、25.00 mg/m L。当药物浓度在1.56 mg/m L及以上时,五倍子对溶藻弧菌生物膜的形成有极显著抑制作用;药物浓度在6.25 mg/m L及以上时,石榴皮对溶藻弧菌生物膜的形成有极显著抑制作用。[结论]五倍子和石榴皮对溶藻弧菌及其生物膜均有抑制作用。     

不同环境因素对表皮葡萄球菌生物膜形成的影响

采用微量板半定量法研究影响表皮葡萄球菌生物膜形成的因素,同时模拟奶牛体内环境因素,如乳糖、酪蛋白和Ca2+等,探索不同理化因素对表皮葡萄球菌生物膜形成的影响,结果表明,检测的3种培养基中,胰蛋白胨大豆肉汤(Typtic Soy Broth,TSB)培养基最有利于表皮葡萄球菌的生长及其生物膜的形成,且最适质量分数为3%左右;在pH中性条件下,NaCl对表皮葡萄球菌生物膜的形成有明显抑制作用;当pH大于7时,表皮葡萄球菌生物膜形成能力明显减弱。在模拟奶牛体内环境因素中,一定质量分数的Ca2+(0.2%及以上)与高质量分数的乳糖(8%及以上)对生物膜形成有明显抑制作用,而外源添加酪蛋白对表皮葡萄球菌生物膜形成没有明显影响。     

An experimental study on nitrification biofilm performances using a series reactor system

Total ammonia nitrogen (TAN) concentration is often a key limiting water quality parameter in intensive aquaculture systems. Removing ammonia through biological filtration is thus the first objective in recirculating aquaculture system design. In this study, the performance characteristics of a steady-state nitrification biofilm were explored using a series of reactors. Four nitrification kinetics parameters were estimated using the data collected from the experimental system, including minimum TAN concentration, half saturation constant, maximum TAN removal rate and maximum specific bacterial growth rate. Experimental data showed that a minimum TAN concentration was needed to support a steady-state nitrification biofilm. For the temperature of 27.2°C, the mean minimum TAN concentration was 0.07 mg/l. For a single substrate-limiting factor, the relationship between TAN removal rate (R) and TAN concentration (S) was represented by an empirical equation [R=1859(S−0.07)/(S+1.93)]. The characteristics of nitrite oxidation were also demonstrated by the experiment system. The results of this study will help to better understand the characteristics of nitrification biofilters applied in recirculating aquaculture systems.     

植物相关细菌生物膜研究进展

介绍了不同类型的植物相关细菌生物膜。综述了有关生物膜结构、形成和微生物与陆生植物结合所形成生物膜的特性等方面的研究进展。阐述了植物表面环境、细菌在植物表面主动和被动的沉积作用等对生物膜形成的影响。     

改进型微动力一体化生物膜反应器处理灰水并回用研究

[目的]为灰水的利用提供依据。[方法]利用改进型微动力一体式生物膜反应器处理灰水,监测了进出水的CODCr、LAS、TN和SS以及反应器的温度、pH值和DO。[结果]进水CODCr平均为188 mg/L,出水CODCr平均为32 mg/L,CODCr去除率为80.2%~86.9%。当水力停留时间(HRT)≤3 h时,系统对污染物的去除率随停留时间的延长增长较快。当HRT>3 h时,系统对污染物的去除率随停留时间的延长增长变缓;HRT=3.5 h时,系统对CODCr、LAS的去除率分别为91.7%、81.2%。反应器在常温下按HRT=3.5 h运行时,处理后灰水符合《生活杂用水水质标准》的要求。CODCr、LAS的去除率随温度变化变化不大,TN的去除率下降了40.4%。[结论]改进型微动力一体化生物膜反应器的最佳水力停留时间为3.5 h,系统对CODCr、LAS的去除率分别为91.7%、81.2%。     

猪场消毒前的准备工作——清洁

2018年发生的非洲猪瘟疫情使得中国养猪业更加注重生物安全,而在养猪生产的生物安全措施中,清洁也是重要的组成部分,是猪场消毒前的重要环节。在猪场的实际工作中,大家往往更加注重消毒而忽略了清洁。文章就猪场清洁工作进行简述。