Characterization by whole-cell hybridization of bacterial populations associated with shrimp hatchery biofilms

The impact of shrimp larvae development, as well as water and food inputs upon the increase of bacterial populations within the bacterial community of hatchery tank biofilms, was studied. For this study, a total of 68 biofilm samples were collected from concrete tanks at three larvae production times in a commercial shrimp hatchery. Seventeen samples were taken at each larval development stage (Zoea I, Mysis I, postlarvae 1 and postlarvae 16), as well as 37 samples from water, shrimp nauplii and food, introduced into the shrimp hatchery tanks. Culturable and direct bacterial counts were performed and 16S‐rRNA‐targeted oligonucleotide probes were used to quantify the presence of specific bacterial groups. An average of 27–70% of DAPI total cell counts were detected with the EUB338 probe, while the GAM42a probe signal ranged from 1% to 11%. Vibrio‐like bacteria (VLB) counts in TCBS agar ranged from <10 to 10¹ VLB/cm⁻², with a tendency to increase at the last postlarvae stage. The most significant external source of bacteria registered with GAM42a probe and TCBS agar were found in live Artemia nauplii, used as food; nevertheless, biofilms remain with low counts of these groups.     

Effects of Dual Microalgal Species Biofilms on New Zealand Black-Footed Abalone (Hailotis iris) Larval/Post-Larval Processes

Dual microalgal species biofilms with distinct amino acid characteristics were tested for their effects on abalone (Haliotis iris) initial attachment, metamorphosis, settlement, and survival. Nine microalgal species, isolated from around Auckland, New Zealand, were grown in the laboratory, and used to produce nine dual microalgal species biofilms. Abalone larvae were exposed to the different biofilm treatments and controls (no biofilms) for seven (attachment and metamorphosis) and 14 (settlement and survival) days. The larvae performed significantly better in the dual microalgal biofilms compared to controls, and some biofilms resulted in better attachment, metamorphosis, settlement, and survival than others. For example, the best-performing biofilms almost always contained cyanobacteria (Oscillatoria cf. erythrata, Anacystis cf. aeruginosa, and Schizothrix cf. calcicola), which are attributed with better nutritional values. However, the amino acid profiles did not produce a clear pattern with regard to their effects on the four larval processes. Instead, total amino acid (TAA) content was positively correlated with percentage of attachment and metamorphosis. Greater TAA contents are likely to reflect greater amounts of extra-cellular polymers within the biofilms, which are suggested to improve larval performance.     

再生水滴灌灌水器附生生物膜生长对堵塞的影响

再生水滴灌灌水器堵塞与其内部附生生物膜的形成和生长密切相关,但是生物膜的生长过程对灌水器堵塞加深的影响机制至今还罕有报道。基于此,该文测试了系统累积运行540 h后7种不同堵塞程度灌水器出流及附生生物膜关键组分,并分析灌水器堵塞加深对生物膜组分增长的敏感性。结果表明:灌水器堵塞程度随着附生生物膜组分的增长而加深,两者存在显著的"S型曲线"相关关系(R20.92),呈现出"敏感-微敏感-极度敏感"变化趋势。生物膜固体颗粒物和磷脂脂肪酸含量变化可以极好地刻画这种敏感性的变化过程,说明水源颗粒物和微生物同时是再生水滴灌灌水器堵塞的关键诱发因素。该文的研究可为揭示再生水滴灌灌水器堵塞诱发机理及其推广提供理论依据。     

鳗鲡循环水高密度养殖试验研究

利用室内封闭式循环水养殖系统对欧洲鳗鲡进行高密度养殖试验。结果表明,10 522尾平均体重为55.6 g(18P)的欧洲鳗鲡养殖159 d,成活率达99.7%,总重由584.6 kg增加到1478.0 kg,均重达143.2 g(7.0P),养殖密度从13.0 kg/m3提高到32.8 kg/m3,共投饵1 263.2 kg,鳗鲡增重893.4 kg,饵料效率达70.7%。采用添加营养液和低负载预培养生物膜,使鳗鲡进入系统后水质平稳变化,降低了养殖初期因水质变化剧烈而发生事故的风险。试验阶段养殖池水体氨氮0.03～1.28 mg/L、亚硝态氮0.02～0.75 mg/L、硝态氮1.21～99.60 mg/L,溶氧控制在5～7 mg/L、pH以碳酸氢钠调节稳定在7.0～7.7、水温在23.8～32.4℃间,系统的日换水量在5%内,各水质指标均处于鳗鲡适宜范围内。养殖期间发生2次指环虫病害,利用中草药和无残留药物进行防治,效果良好。利用循环水养殖系统养殖鳗鲡,创造最适的水环境理化条件,在快速生产绿色安全水产品的同时有效节水和减少污水排放。研究亮点:国内首次中试规模(养殖水体45 m3),高密度(32.8 kg/m3)进行了欧洲鳗鲡的循环水养殖试验。养殖试验时间达159 d,鳗鲡达到了商品规格。在中试规模条件下,联合运用预培养生物膜和低负载培养生物膜的方法快速构建了具有稳定硝化功能的生物过滤器。首次比较了不同日换水率条件下,循环水养殖欧洲鳗鲡水体氨氮、亚硝酸盐氮和硝酸盐氮的变化情况。     

被膜态与浮游态猪源乳杆菌上清液抑菌活性比较分析

试验旨在比较分析猪源约氏乳杆菌L-76和嗜淀粉乳杆菌L-102被膜态和浮游态下其上清液对病原菌的抑菌活性。采用结晶紫染色法测定L-76和L-102的生物被膜形成能力,研究其被膜态和浮游态乳杆菌上清液及上清液与不同影响因素作用后的抑菌活性,并利用扫描电镜观察其两种状态下的上清液对指示菌形态的影响。结果显示,L-76、L-102乳酸菌具有较强的生物被膜形成能力,被膜态和浮游态乳杆菌L-76、L-102具有良好的抑菌活性,其上清液经蛋白酶作用后,被膜态L-76和L-102上清液的抑菌圈直径比浮游态明显变小;经过氧化氢酶和不同温度作用后,两种状态的乳杆菌上清液的抑菌活性无明显变化（P>0.05）,但在pH 3.0作用下,两种状态的乳杆菌上清液的抑菌效果最好;扫描电镜结果显示,被膜态乳杆菌上清液对金黄色葡萄球菌的形态影响略大。综上所述,两种状态下乳酸杆菌上清液中均含有抑菌物质,且被膜态乳酸杆菌上清液中的抑菌物质含量略多或活性略高。     

Biofilm production by pathogens associated with canine otitis externa,and the antibiofilm activity of ionophores and antimicrobial adjuvants

Otitis externa (OE) is a frequently reported disorder in dogs associated with secondary infections by Staphylococcus, Pseudomonas and yeast pathogens. The presence of biofilms may play an important role in the resistance of otic pathogens to antimicrobial agents. Biofilm production of twenty Staphylococcus pseudintermedius and twenty Pseudomonas aeruginosa canine otic isolates was determined quantitatively using a microtiter plate assay, and each isolate was classified as a strong, moderate, weak or nonbiofilm producer. Minimum biofilm eradication concentration (MBEC) of two ionophores (narasin and monensin) and three adjuvants (N‐acetylcysteine (NAC), Tris‐EDTA and disodium EDTA) were investigated spectrophotometrically (OD_570nm) and quantitatively (CFU/ml) against selected Staphylococcus and Pseudomonas biofilm cultures. Concurrently, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of planktonic cultures were assessed. 16/20 of the S. pseudintermedius clinical isolates were weak biofilm producers. 19/20 P. aeruginosa clinical isolates produced biofilms and were distributed almost equally as weak, moderate and strong biofilm producers. While significant antibiofilm activity was observed, no MBEC was achieved with narasin or monensin. The MBEC for NAC ranged from 5,000–10,000 µg/ml and from 20,000–80,000 µg/ml against S. pseudintermedius and P. aeruginosa, respectively. Tris‐EDTA eradicated P. aeruginosa biofilms at concentrations ranging from 6,000/1,900 to 12,000/3,800 µg/ml. The MBEC was up to 16‐fold and eightfold higher than the MIC/MBC of NAC and Tris‐EDTA, respectively. Disodium EDTA reduced biofilm growth of both strains at concentrations of 470 µg/ml and higher. It can be concluded that biofilm production is common in pathogens associated with canine OE. NAC and Tris‐EDTA are effective antibiofilm agents in vitro that could be considered for the treatment of biofilm‐associated OE in dogs.     

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.     

Anti-biofilm compounds derived from marine sponges

Bacterial biofilms are surface-attached communities of microorganisms that are protected by an extracellular matrix of biomolecules. In the biofilm state, bacteria are significantly more resistant to external assault, including attack by antibiotics. In their native environment, bacterial biofilms underpin costly biofouling that wreaks havoc on shipping, utilities, and offshore industry. Within a host environment, they are insensitive to antiseptics and basic host immune responses. It is estimated that up to 80% of all microbial infections are biofilm-based. Biofilm infections of indwelling medical devices are of particular concern, since once the device is colonized, infection is almost impossible to eliminate. Given the prominence of biofilms in infectious diseases, there is a notable effort towards developing small, synthetically available molecules that will modulate bacterial biofilm development and maintenance. Here, we highlight the development of small molecules that inhibit and/or disperse bacterial biofilms specifically through non-microbicidal mechanisms. Importantly, we discuss several sets of compounds derived from marine sponges that we are developing in our labs to address the persistent biofilm problem. We will discuss: discovery/synthesis of natural products and their analogues-including our marine sponge-derived compounds and initial adjuvant activity and toxicological screening of our novel anti-biofilm compounds.     

两级回流生物膜工艺处理农村生活污水效果

针对农村生活污水低碳高氮磷特点,研究两级回流连续曝气生物膜工艺处理农村生活污水的效果和机理。在平均处理量90t/d、水力停留时间为3.0d,稳定运行12个月结果表明,该工艺对化学需氧量、五日生化需氧量、氨氮、全氮、全磷和悬浮物的平均去除率分别为75.7%、84.8%、69.2%、68.0%、58.9%和87.9%;出水化学需氧量、五日生化需氧量、氨氮、全氮、全磷和悬浮物的平均质量浓度分别在51.0、15.8、10.8、16.5、2.3和15.7mg/L以下,满足天津市农村污水处理厂排水标准,改造后工艺运行更加稳定,出水NH3-N和TN有明显改善,符合农村污水处理要求。     

蜡样芽孢杆菌及其生物被膜对有机酸及乙醇的抗性比较

为比较蜡样芽孢杆菌及其生物被膜对环境压力的抗性,该文主要研究了4种有机酸（乙酸、柠檬酸、乳酸和苹果酸）和乙醇对蜡样芽孢杆菌及其生物被膜存活率的影响。结果表明：生物被膜态蜡样芽孢杆菌对乙酸的抗性高于浮游态菌。扫描电镜结果显示,经乙酸处理后,浮游态蜡样芽孢杆菌的细胞表面严重受损,而生物被膜态菌的表面形态未发生明显变化。在柠檬酸、乳酸、苹果酸和乙醇存在的条件下,生物被膜态蜡样芽孢杆菌比浮游态菌表现出更强的压力抗性,特别是在高浓度（有机酸16%～20%,乙醇50%～60%）的情况下,该现象尤为显著。因此,在食品工业中控制被膜态蜡样芽孢杆菌对预防和阻止食品腐败非常重要。该研究结果为实际生产中有效控制蜡样芽孢杆菌及其生物被膜的形成提供参考。