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.     

The impact of temperature on nitrification rate in fixed film biofilters

The impact of temperature on nitrification rate was evaluated in this study through experimental tests, mathematical modeling and sensitivity analysis. The results show that the impact of temperature on fixed film nitrification rate is less significant than that predicted by the van't Hoff–Arrhenius equation. In a fixed film biofilter, the impact of temperature on nitrification rate due to DO (dissolved oxygen) limitation is different from that due to TAN (total ammonia nitrogen) limitation. Sensitivity analysis indicated that a temperature increment at 20 °C resulted in nitrification rate increase of 1.108% per °C and 4.275% per °C under DO and TAN limited conditions, respectively. Diffusion mass transport plays an important role in fixed film nitrification processes. Consequently, the effect of temperature on nitrification rate due to bacterial growth rate change in fixed film processes is greatly reduced compared with that of suspended growth processes. When oxygen is limited, the decrease in saturation DO concentration as temperature increases results in a negative temperature impact upon the nitrification rate.     

Larval settlement of the silver- or goldlip pearl oyster Pinctada maxima (Jameson) in response to natural biofilms and chemical cues

This paper presents the first laboratory study investigating larval settlement and metamorphosis of Pinctada maxima in response to both natural and artificial inducers. Natural biofilms induced larval settlement and the inductive effect appears to be closely associated with the bacteria in the biofilm. This result indicates that, in nature, bacteria may play an important role in the settlement of P. maxima larvae. Various pharmacological agents were also tested in this study and the results were generally similar to those found with other marine invertebrates, particularly molluscs. The results show that some pharmacological agents are potent artificial inducers of larval settlement for P. maxima. The findings of this study provide valuable information for understanding the mechanism of the larval settlement of this species, which is useful information for the hatchery industry of this pearl oyster.     

Nitrification kinetics of biofilm as affected by water quality factors

Various types of fixed film biofilters have been used in recirculating aquaculture systems under different water quality and operating conditions. The effectiveness of the nitrification process can be evaluated by nitrification kinetics. Nitrification in the bacterial film of the biofilter involves physical, chemical and biological processes that are governed by a variety of parameters such as substrate and dissolved oxygen concentrations, organic matters, temperature, pH, alkalinity, salinity and turbulence level. The impacts of these parameters upon nitrification kinetics make predicting the performance of a biofilter for a given application an engineering challenge. Knowing the performance of a biofilter is critical for both designers and managers. This paper summarizes the current knowledge on nitrification kinetics as affected by the aforementioned factors based on literature and the results from the authors’ laboratories. These factors were ranked according to their significance of impact on biofilter nitrification performance. The information presented can be used as a reference for the design and operation of biofilters in recirculating aquaculture systems.     

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。     

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。     

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.     

循环水养殖系统生物滤器负荷挂膜技术

循环水养殖系统启动运行前往往需要经过一段时间的生物膜预培养,使生物膜达到成熟稳定,从而保证系统的水质净化功能。本研究通过养殖试验,研究了生物滤器负荷挂膜的技术方法,以期实现生物膜的快速成熟和系统的快速启动。为此,构建了6组循环水系统组成的养殖车间,建成后立即投入试验生产。试验为期120 d,养殖种类为红鳍东方鲀,初始放养平均体重(632.5±2.26)g。期间,红鳍东方鲀平均增重29.91%,养殖成活率98.7%,养殖密度由(19.34±1.89)kg/m3增加到(32.17±3.40)kg/m3,投饵率由0.2%增加到0.5%–0.7%,每日换水量由50%逐渐减至10%。结果表明,在生物膜的生长期,通过对投饵量及新水补充量的有效调节,可以把养殖水体中的氨氮和亚硝氮浓度控制在安全范围以内,以保证养殖鱼类的生长。生物膜在50天左右达到完全成熟,此后便可依靠生物膜的净化作用将氨氮浓度控制在0.5?1.2 mg/L、亚硝氮浓度控制在0.2?0.5 mg/L、pH值控制在6.5–7.5、COD值低于4 mg/L、细菌总数控制在800–2100 cell/ml的安全范围内。利用生物滤器负荷挂膜技术,在合理调控水质指标的条件下,循环水养殖系统建成后可以立即投入生产,实现生物滤器挂膜与养殖生产的同步进行。