生物强化移动床生物膜反应器处理水产养殖循环水初步研究

利用自制的硝化细菌菌剂促进移动床生物膜反应器(Moving bed biofilm reactor,MBBR)的挂膜启动,分析不同载体氨氮负荷、碳氮比条件下反应器运行状况,并进一步进行了实验室模拟循环水养殖草金鱼实验。结果显示,利用自制硝化菌剂能够完成整个移动床反应器的启动过程,在接种15 d后使循环出水氨氮稳定在1 mg/L以下。单位体积载体氨氮负荷实验表明,MBBR能够在100 mg TAN/(L填料·d)条件下,使出水满足一般水产养殖水质要求(氨氮0.5 mg/L,亚硝氮0.1 mg/L)。进水碳氮比在1以内时MBBR能够稳定高效运行。在实验室模拟循环水养殖过程中,经菌剂强化的MBBR能维持循环出水氨氮低于0.5 mg/L,亚硝氮低于0.05 mg/L。     

A case study of biofilter activation and microbial nitrification in a marine recirculation aquaculture system for rearing Atlantic salmon (Salmo salar L.)

Marine recirculation aquaculture system (RAS) is a prominent technology within fish farming. However, the nitrifying bacteria in the biofilter have low growth rates, which can make the biofilter activation a long and delicate process with periods of low nitrification rates and variations in water quality. More knowledge on the microbial development in biofilters is therefore needed in order to understand the rearing conditions that favour optimal activation of the biofilters. In this case study, we investigated the activation of two biofilters in a marine RAS for Atlantic salmon post‐smolt associated with either high or low stocking densities of fish by monitoring the microbial communities and chemical composition. The results showed that the microbial communities in both biofilters were similar during the first rearing cycle, despite variations in the water quality. Nitrifying bacteria were established in both biofilters; however, the biofilter associated with low stocking density had the highest relative abundance of ammonia‐oxidizing Nitrosococcus (1.0%) and nitrite‐oxidizing Nitrospira (2.1%) at the end of the first rearing cycle, while the relative abundance of ammonia‐oxidizing Nitrosomonas (2.3%–2.9%) was similar in both biofilters. Our study showed that low fish stocking density during the first rearing cycle provided low and steady concentrations of ammonium, nitrite and organic load, which can stimulate rapid development of a nitrifying population in new marine RAS biofilters.     

循环海水养殖系统硝化滤器中氨氧化微生物分析

研究循环水养殖硝化滤器载体上附着生物膜的微生物群落结构可以为提高其处理速率和效率,并为特异性工程菌构建提供依据。采用改良的AFLP方法分析了循环水养殖硝化滤器载体上附着的氨氧化细菌16S rRNA基因和氨单加氧酶amoA基因片段及其系统发育情况。结果表明:分析16S rRNA基因得到的序列片段比分析amoA基因片段得到了更多信息,准确度较高,可作为分析循环水养殖硝化滤器氨氧化菌群组成的有效方法。克隆测序所得序列与网上公布数据比对,可见存在于循环水养殖硝化滤器载体上的氨氧化细菌与Nitrosomonas cryotolerans、Nitrosomonas oligotropha、Nitrosospira tenuis、Nitrosomonas marina相似度达100%,与Nitrosomonas aestuarii相似度为87%。大部分属于亚硝化单胞菌属(Nitrosomonas),仅少数序列属于亚硝化螺菌属(Nitrosospira)。采用16S rRNA基因和amoA片段分析方法得到的附着于封闭循环海水养殖硝化滤器载体上的氨氧化细菌主要为变形菌(Proteobacteria)的β-亚类的亚硝化单胞菌属(Nitrosomonas)和少量的亚硝化螺菌属(Nitrosospira)氨氧化细菌,以及一定数量的γ-亚类氨氧化细菌。     

Effects of formaldehyde on nitrification in biofilters of small‐scale recirculating systems

Florfenicol (Aquaflor®) is the only U.S. Food and Drug Administration (FDA) approved drug for treating diseased fish reared in recirculating aquaculture systems (RAS). Treating diseased fish in RAS is challenging because of the potential to damage nitrifying bacteria in the biofilters. Impaired nitrification can lead to concentrations of ammonia and nitrite that compromise fish welfare. The objective of this study was to determine the effects of a FDA‐approved parasiticide and fungicide, Parasite‐S^® (formalin), on biofilter nitrification. Stable biofilters were exposed once to 0, 9.25, 18.5, 37, or 55.5 mg/L formaldehyde. Total ammonia nitrogen (TAN) and nitrite nitrogen were monitored daily before and throughout the study to quantify biofilter function. Formaldehyde concentrations ≥37 mg/L increased TAN and nitrite nitrogen concentrations, and nitrification did not recover to pre‐exposure concentrations up to 8 day postexposure. On the basis of those results, a second trial was conducted. Stable biofilters were exposed once or on four consecutive days to 9.25 or 18.5 mg/L formaldehyde. Biofilters repeatedly exposed to formaldehyde showed signs of impairment and had variable recovery relative to single exposures. Results of this study may help identify formaldehyde concentrations that can be safely applied to RAS when treating diseased fish.     

Variability of the nitrifying bacteria in the biofilm and water column of a recirculating aquaculture system for tilapia (Oreochromis niloticus) production

The aim of this study was to evaluate variability of nitrifying bacterial community in the biofilm and in the water of a recirculating aquaculture systems (RAS) in a tilapia farming in order to determine if nitrification process is dependent, or not, of nitrifying bacteria abundance. Biofilm and water samples were collected periodically for 30 days and analysed with the fluorescent in situ hybridization (FISH) technique, used to quantify ammonia‐oxidizing bacteria (AOB) and nitrite‐oxidizing bacteria (NOB). Ammonia presented the peak in the first week, while the nitrite's maximum was recorded in the second week. Nitrate increased steadily, indicating nitrification activity. Total bacterial abundance in biofilm increased continuously, while in water, it did not change significantly. In the biofilm, number of AOB was high at beginning, decreased after few days and increased again following augment of ammonia. Number of NOB also showed an increase in abundance in biofilm following the increment of nitrite and nitrate. In water, AOB and NOB did not show major variability. Relative abundance of nitrifying bacteria represented more than 30% of total bacteria in biofilm at beginning of the experiment. Their contribution decreased to >3% in last days. It indicates that nitrifying bacteria are biofilm colonizers, and that their activity seems to be directly related to the concentration of nitrogen compounds. However, contribution of nitrifying bacteria did not vary much along the time. We may conclude that the biofilm‐nitrifying bacteria plays major role in nitrification process in RAS and that the activity of these organisms is dependent of their abundance in response to the concentration of nitrogen compounds.     

Biocontrol of ammonia pollution in the rearing water of fish by inducing a heterotrophic bacterial-based food chain in the medium

In intensive aquaculture, one of the main problems confronted by the farmers is the ammonia pollution and subsequent disease outbreaks, high costs of quality protein feed, and the labor for periodic water exchange. Ammonia is a major metabolic waste product from fish, which is excreted across the gill membranes and in the urine. Controlling the inorganic nitrogen by manipulating the carbon/nitrogen ratios seems to be a practical and inexpensive means of reducing the accumulation of ammonia. At high carbon to nitrogen (C/N) ratios, bacteria will assimilate nitrogen, i.e., ammonia, from water and produce new cell protein. The experiment was designed by adding 0, 10, 20, and 30 g of carbohydrate (rice flour) for each gram of total ammonia nitrogen (TAN) released as a result of feeding metabolism and feed waste decomposition. The ammonia built up in the rearing water showed a drastic decrease in all the carbohydrate added tanks. The heterotrophic bacterial growth was significantly higher in the same. The biochemical constituents and growth rate were higher in fishes in the tanks having C/N ratio 10 and 20. Percentage weight gain was 100% for T₃₀, protein efficiency ratio (PER) was high for T₂₀ and T₃₀ (4.048). The C/N ratios of 20 and 30 worked more effectively. Shifting the aquatic ecology from autotrophic to heterotrophic bacterial-based community can improve water quality and recycle the toxic ammonia waste to heterotrophic bacterial flocs, which in turn can be consumed by the fish, thereby reducing the feed protein demand and subsequently the reduction of feed cost.     

不同有机碳源及C/N对生物滤池净化效果的影响

利用生物滤池模拟装置,以实际养殖废水为处理对象,探讨了4种常见有机碳源(葡萄糖、乙醇、红糖和淀粉)及不同碳氮比对有机物去除、硝化反应和异养反硝化作用等生物滤池主要净化过程的影响.碳源初选结果显示,同种碳源下,当C/N从0升高至6过程中,生物滤池对TAN(总氨氮)的去除率呈先升高后降低趋势;当C/N较小时,各组对NO2--N的去除率差异性不显著(P＞0.05),随着C/N继续升高,NO2-N去除率则显著降低(P＜0.05);乙醇组除外,其他3组随着C/N升高,CODMn去除率先迅速增大然后趋于稳定;各组NO3-N和TN去除率呈先升高后降低趋势,且变化显著(P＜0.05),当C/N=4时,分别达到最高值.碳源复选结果显示,在C/N=4条件下,分别添加有机碳源(乙醇、淀粉、红糖和葡萄糖)的4组对TAN、NO3--N、TN和CODM的去除率显著高于对照组(P＜0.05);而对照组NO2--N的去除率最高,达到93.59％;添加乙醇,生物滤池对水体中TAN、NO2-N、NO3-N和TN的去除效果优于其他3种碳源.研究表明,当C/N=4时,乙醇作为外加碳源能很好地提高生物滤池的净化效率.     

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.     

Development of a Model to Simulate Nitrogen Dynamics in an Integrated Shrimp–Macroalgae Culture System with Zero Water Exchange

The effluents of traditional shrimp monoculture cause pollution and promote eutrophication and hypernutrification of the receiving coastal ecosystems. Integrated aquaculture and a recirculating aquaculture system (RAS) have been proposed as an alternative to address these problems. In this study, we developed a dynamic model to simulate the concentration of total ammonia nitrogen (TAN), nitrite, and nitrate in an integrated culture of whiteleg shrimp, Litopenaeus vannamei, and seaweed, Gracilaria vermiculophylla, in a recirculating and zero water exchange system, and the effect of nitrifying and heterotrophic bacteria was also included. The experiments demonstrated that a dynamic model can explain the concentrations of dissolved inorganic nitrogen and variations in these concentrations over time in the integrated culture. The results also suggest that nitrifying and heterotrophic bacteria play an important role in the transformation of dissolved nitrogenous compounds; therefore, these bacteria should be considered within the dynamics of nitrogen in integrated systems with low water exchange.