The prevention of microfouling and macrofouling on hydrogels impregnated with either Arquad 2C-75 or benzalkonium chloride

Optically clear, surfactant loaded poly (2-hydroxyethyl methacrylate) (pHEMA) hydrogels can be used to prevent fouling on optical windows of marine underwater sensors. To act successfully in this capacity, hydrogels need to prevent both microfouling and macrofouling. Panel trials were conducted using four different materials: unloaded hydrogels, hydrogels containing either benzalkonium chloride (BAC) or dicocodimethylammonium chloride (Arquad 2C-75) and PMMA coupons. Three panels were deployed at staggered intervals (2, 4 and 6 weeks) before the main settlement season of Semibalanus balanoides and Mytilus edulis in the Firth of Clyde, Scotland. Panels were left for a total period of 10, 12 and 14 weeks respectively. Results showed that no sample completely resisted fouling, but Arquad 2C-75 hydrogels were extremely effective at preventing both microfouling and macrofouling. The most heavily fouled materials were unloaded hydrogels and PMMA, despite differences in initial hydrophilicities. Arquad 2C-75 hydrogels were equally effective at preventing larval settlement, for up to 14 weeks.     

The prevention of microfouling and macrofouling on hydrogels impregnated with either Arquad 2C-75® or benzalkonium chloride

Abstract

Optically clear, surfactant loaded poly (2-hydroxyethyl methacrylate) (pHEMA) hydrogels can be used to prevent fouling on optical windows of marine underwater sensors. To act successfully in this capacity, hydrogels need to prevent both microfouling and macrofouling. Panel trials were conducted using four different materials: unloaded hydrogels, hydrogels containing either benzalkonium chloride (BAC) or dicocodimethylammonium chloride (Arquad 2C-75®) and PMMA coupons. Three panels were deployed at staggered intervals (2, 4 and 6 weeks) before the main settlement season of Semibalanus balanoidesand Mytilus edulisin the Firth of Clyde, Scotland. Panels were left for a total period of 10, 12 and 14 weeks respectively. Results showed that no sample completely resisted fouling, but Arquad 2C-75® hydrogels were extremely effective at preventing both microfouling and macrofouling. The most heavily fouled materials were unloaded hydrogels and PMMA, despite differences in initial hydrophilicities. Arquad 2C-75® hydrogels were equally effective at preventing larval settlement, for up to 14 weeks.     

In situ identification of bacterial species in marine microfouling films by using an immunofluorescence technique

An immunofluorescence technique was developed for the in situ identification of specific bacteria in marine microfouling films. Microorganisms adherent to glass plates after 30 days of immersion in a synthetic seawater system were cultured and classified by biochemical tests, flagellar arrangement, and the API 20E system. All isolates were gram-negative aerobic or facultative motile rods, predominantly Pseudomonas spp. Rabbit antisera to the five dominant organisms including Achromobacter spp., Comamonas terrigena, P. putrefaciens, a yellow-pigmented Pseudomonas sp., and Vibrio alginolyticus were prepared. These antisera were shown to be species specific in indirect immunofluorescence assays against a battery of 26 marine isolates from 14 bacterial species, with the exception of antisera to the Pseudomonas spp, which cross-reacted with each other but not with test bacteria of other genera. These immunofluorescent reagents enabled the in situ identification of all five bacterial species in microfouling films. Low-surface-energy test plates had smaller numbers of adherent bacteria in microfouling films than medium-surface-energy test plates, suggesting that the degree of microfouling may be influenced by the surface energy. In addition, the reagents could identify up to 39% of the attached bacteria in microfouling films spontaneously formed on steel plates in flow cells deployed in different areas of the Atlantic Ocean. The microbial composition of the ocean-formed films varied with the geographical area of their formation. The present results indicate that immunofluorescence techniques may provide a rapid and reliable means to identify, in situ, specific bacteria in marine microfouling films.     

Effect of Manual Brush Cleaning on Biomass and Community Structure of Microfouling Film Formed on Aluminum and Titanium Surfaces Exposed to Rapidly Flowing Seawater

Metals exposed to rapidly flowing seawater are fouled by microbes that increase heat transfer resistance. In this study, results of biochemical test methods quantitatively relating the biomass and community structure of the microfouling film on aluminum and titanium to heat transfer resistance across the metal surface during three cycles of free fouling and manual brushing showed that cleaning accelerates the rate of fouling measured as the loss of heat transfer efficiency and as microfouling film biomass. The results also showed that the rate of fouling, measured as an increase in heat transfer resistance, is faster on titanium than on aluminum but that the titanium surface is more readily cleaned. In three cycles of free fouling and cleaning with a stiff-bristle nylon brush, the free-fouling communities re-forming on aluminum became enriched in bacteria containing short-branched fatty acids as the cycling progressed. The free-fouling community on titanium revealed an increasingly diverse morphology under scanning electron microscopy that was enriched in a portion of the microeucaryotes. Brushing removed most of the biomass, but left a residual community that was relatively enriched in a portion of the bacterial assembly containing cyclopropane fatty acids on aluminum and in a more diverse community on the titanium surface. The residual communities left after cleaning of titanium revealed an increase in bacteria with short-branched fatty acids and in microeucaryotes as cleaning continued. No significant changes occurred in the residual microbial community structure left on aluminum with cleaning; it was, again, less diverse than that remaining on titanium. The residual communities secreted a twofold-larger amount of extracellular polymer, measured as the ratio of total organic carbon to lipid phosphate, than did the free-fouling community on both surfaces.     

The use of periphytic diatoms as a means of assessing impacts of point source inorganic nutrient pollution in south-eastern Australia

1. Periphytic diatoms are used as indicators of water quality because their ecological tolerances or preferences to environmental variables are thought to be predictable. However, much of the present autecological information for periphytic diatoms has been derived from studies conducted in the northern hemisphere. In this present study we used periphytic diatoms to determine the impacts of inorganic nutrient pollution in a tidal river system in the temperate latitudes of south‐east Australia. In so doing, we assess the suitability of the use of the ‘northern hemisphere’ ecological tolerance/preference data for periphytic diatoms. 2. Artificial substrates were used to collect periphytic diatoms at 35 sites, which were positioned along the riverbanks and the middle of the river at various distances upstream and downstream of the sewage outfall. The sampling design took into account tidal excursions and the observed sewage plume dynamics. Periphytic diatoms were collected during the austral winter month of August and the austral spring months of September and October. We deployed the artificial substrates for 4 weeks to allow the periphytic diatoms to recruit and colonise, before identifying and enumerating the assemblages. 3. Data analysis included two approaches: multivariate visualisations of combinations of environmental and biological data to investigate shifts in species structure of the periphytic diatom assemblage and multimetric indices based on ecological tolerance/preference data. 4. We found that the spatial patterns inferred from multivariate and multimetric analyses were consistent. Temporal variation in the composition of the periphytic diatom assemblage was greater than the spatial variation along horizontal sections of the river (in any one deployment) due mainly to shifts between winter and spring species. 5. Outfall effects were most apparent in winter, possibly because subsequent deployments were swamped by growth of spring periphytic diatoms. The outfall effects included a shift towards pollutant tolerant species and a reduction in the variability of the periphytic diatom assemblage across the river. 6. We conclude that the use of periphytic diatoms and associated ecological tolerance/preference data as a means of assessing impacts of point source inorganic nutrient pollution is effective. An understanding of river and sewage flow patterns is essential to the design of appropriate monitoring programmes and to the interpretation of results, especially as periphytic diatoms are sensitive to many environmental variables.     

Influence of substrate type on periphyton biomass and nutrient state at contrasting high nutrient levels in a subtropical shallow lake

How substrate affects periphyton biomass and nutrient state at different, but high, nutrient levels was tested in three large enclosures in a hypereutrophic subtropical shallow lake. We compared periphytic characteristics (1) on three hard substrates (stone, bamboo, and wood) incubated for 2 weeks and 1 year, respectively, to investigate the existence of the influences of substrate type at hypereutrophic levels, and (2) on artificial plants with contrasting (parvopotamid-like and myriophyllid-like) soft substrate morphology. In general, periphytic biomass and nutrient state were sensitive to variations in nutrient level, incubation time, hard substrate type (except 2-week incubated) and substrate morphology, but to a varying extent. The periphyton nutrient content increased with increasing nutrient levels on most substrates. Long-time incubated substrates supported more periphytic biomass, had a higher nutrient content and autotrophic proportion, while the effect of nutrient level on nutrient content in the periphyton was independent of incubation time. The effects of hard substrate type on periphyton characteristics were much weaker than those of nutrient level. By contrast, the effects of soft substrate morphology on periphyton biomass and carbon: nutrient ratios surpassed those of nutrient level. Chlorophyll a, dry mass, and ash free dry mass were much higher on parvopotamid than on myriophyllid substrates. Our results show that periphyton biomass and nutrient state are influenced by both substrate and nutrient level even in hypereutrophic lakes, which might have cascading effects on the benthic food web.     

空间层次及人工基质对着生藻类建群特性的影响

为全面了解着生藻类在建群中群落变化的生态学特性,揭示着生藻类的建群规律,在以丝状藻类为优势藻的生态塘中,采用花岗岩和瓷砖为附着材料,设置水体底部和中部为附着位点,进行频次为10d的采样分析。结果表明,生态塘中共检出8门73属117种着生藻类,其中以硅藻、蓝藻、绿藻为优势类群。同时不同人工基质和不同空间层次条件下着生藻类的建群特征较一致,早期以单细胞硅藻如舟形藻(Navicula sp.)、脆杆藻(Fragilaria sp.)、曲壳藻(Achnanthes sp.)等为优势,后期以丝状藻类如鞘丝藻(Lyngbya sp.)、颤藻(Oscillatoria sp.)、伪鱼腥藻(Pseudanabaena sp.)等为优势;研究结果发现不同人工基质(花岗岩和瓷砖)对着生藻类的种类组成、细胞密度、生物量和藻类多样性无显著影响,花岗岩和瓷砖上附着的着生藻类具有较高的相似性;但不同的空间层次对着生藻类建群特征影响明显,水体底部具有更多的硅藻种类数,中部具有更多的绿藻,随着建群时间的发展,蓝藻比例不断增加;就生物量而言,底部的着生藻类叶绿素a显著高于水体中部,但两者的细胞密度无显著性差异;随着建群过程的发展,水体底部的着生藻类生物量达峰值所需的时间比中部更长。通过相关性分析,生态塘中着生藻类的生长主要受总磷的影响。     

Quantification of the geocarposphere and rhizosphere effect of peanut (Arachis hypogaea L.)

Roots and pods of field-grown peanut (groundnut) (Arachis hypogaea L.) were sampled at the R3, R5, and R7 developmental stages and examined in comparison to root- and pod-free soil for microbial population densities to assess the geocarposphere and rhizosphere effects. G/ S (no. geocarposphere microorganisms/no. soil microorganisms) and R/S (no. rhizosphere microorganisms/no. soil microorganisms) ratios were calculated for total fungi,Asperigillus flavus, spore-forming bacilli, coryneform bacteria, fluorescent pseudomonads, and total bacteria isolated on low- and high-nutrient media. A clear geocarposphere effect was evidenced by increased population densities of bacteria and fungi associated with developing pods compared to soil. G/S and R/S ratios were generally greater than 1.0 for all groups of microorganisms except bacilli. G/S ratios were greater for total bacteria than for total fungi at two of the three sample times, suggesting that bacteria were stimulated more than fungi in the zone around developing pods. In contrast, R/S ratios, were higher for total fungi than for total bacteria at two of three sample times. The preferential association of fungi and bacteria with early developmental stages of the pod indicates that some microorganisms are particularly well adapted for colonization of the peanut geocarposphere. These microorganisms are logical candidates for evaluation as biological control candiates forA. flavus.     

雪被去除对川西高山冷杉林冬季土壤微生物生物量碳氮和可培养微生物数量的影响

为了解气候变暖情景下雪被减少对冬季土壤微生物特征的影响,采用人工遮雪的方法,研究了雪被去除对原始冷杉林土壤微生物生物量和可培养微生物数量的影响.结果表明:雪被去除显著影响土壤微生物生物量碳(MBC)和氮(MBN)以及可培养细菌和真菌数量,但土壤微生物在雪被覆盖不同阶段具有不同的响应特征.在雪被去除处理下,土壤有机层MBC和MBN在雪被形成初期和雪被融化前期显著降低,而在雪被覆盖期和雪被融化后期显著增加;在雪被形成初期至雪被覆盖期,可培养细菌数量都显著降低,但可培养真菌数量都显著增加.雪被融化后,雪被去除显著降低土壤有机层MBC和可培养真菌数量,显著增加可培养细菌数量,对MBN无显著影响.矿质土壤层MBC、MBN和可培养微生物数量在雪被去除下的变化趋势与土壤有机层基本一致,但波动较小.雪被去除还改变了川西高山冷杉林冬季土壤微生物类群比,提高了土壤可培养真菌数量的冬季优势.     

Multi-Etiological Nature of Tuberculosis-Like Lesions in Condemned Pigs at the Slaughterhouse

Tuberculosis-like lesions (TBL) in pigs have been associated with microorganisms other than mycobacteria. In this work a histopathological and microbiological evaluation of TBL in pigs is shown. A total of 352 samples belonging to 171 pigs totally condemned at slaughterhouse due to generalized TBL were sampled and selected for analysis. Pyogranulomatous (56.2%) and granulomatous lesions (20.2%) were observed in all analysed organs. Most of the granulomas observed in both lymph nodes and lungs belonged to more advanced stages of development (stages III and IV) whereas in the liver and the spleen most of lesions belonged to intermediate stages (stages II and III). Different microorganisms were simultaneously detected from TBL in the 42.7% of the animals. Mycobacterium tuberculosis complex (MTC) (38%), coryneform bacteria (40.3%) and streptococci (28.1%) were the main groups of microorganisms detected after bacteriological analysis, with Trueperella pyogenes and Streptococcus suis as the most frequently isolated species. Mycobacteria belonging to MTC were the most frequently detected pathogens in granulomatous and pyogranulomatous lesions in submandibular lymph nodes (32.7%) and coryneform bacteria were the microorganisms more frequently isolated from lungs (25.9%) and spleen samples (37.2%). These results may provide new insights into the pathogenesis and diagnosis of this pathology. The importance of coryneform bacteria and streptococci in such processes must be evaluated in future studies.     

Chrysophyte stomatocyst assemblages associated with periphytic, high arctic pond environments

Chrysophycean stomatocysts associated with three different periphytic substrates (wet mosses, submerged mosses and rock scrapes) were investigated from ponds on Cape Herschel (78°37'N, 74°42'W), Ellesmere Island in the Canadian High Arctic. The goal of this study was to determine whether a distinct assemblage of periphytic chrysophyte cysts existed and, if so, whether assemblage composition varied with substrate and between ponds. One hundred and thirty-seven different cyst morphotypes were observed with light microscopy from 68 periphytic samples taken from 35 ponds. Twenty-six of these cysts were new morphotypes, of which 16 were identified and described using scanning electron microscopy. Significantly more cyst types with collars and hooked projections in the collar region (i.e. 'hooked'), and fewer unomamented morphotypes were recorded in the periphytic habitats as compared to the surface sediments. Wet moss stomatocyst assemblages were particularly distinct, with a high number of heavily silicified and hooked morphotypes. The morphotype richness was far greater in periphytic environments, with 86, 100 and 95 morphotypes observed in the wet mosses, submerged mosses and rock scrapes, respectively, as compared to only 35 types in the surface sediments of the ponds. Canonical correspondence analysis indicated that measured water chemistry did not account for the variation in the species data (p_{axic 1} < 0.01, 999 Monte Carlo permutations). This study suggests that distinct periphytic assemblages exist, and that cyst morphotype composition and richness varies with substrate.     

Non-chemical biofouling control in heat exchangers and seawater piping systems using acoustic pulses generated by an electrical discharge

Acoustic pulses generated by an electrical discharge (pulsed acoustics) were investigated as a means for biofouling control in two test formats, viz. a 5/8" outside diameter titanium tube and a mockup heat exchanger. The pulsed acoustic device, when operated at 17 kV, demonstrated 95% inhibition of microfouling over a 20 ft length of titanium tube over a 4-week period, comparable to chlorination in combination with a high-velocity flush. The pulsed acoustic device inhibited microfouling over a downstream distance of 15 ft, therefore, a single pulsed acoustic device is theoretically capable of protecting at least 30 ft of tube from microfouling (15 ft on either side of the device). A correlation between acoustic intensity in the frequency range 0.01-1 MHz and the level of biofouling inhibition was observed. The threshold acoustic intensity for microfouling inhibition was determined for this frequency range. It was also observed that the orientation of the device is critical to obtaining microfouling inhibition.     

Influence of substrate wettability on the attachment of marine bacteria to various surfaces

The effect of the initial substrate surface condition, as indicated by the critical surface tension for wetting, on the rate of attachment of marine bacteria to a variety of solid surfaces has been measured. The techniques used to determine the number of bacteria attached per unit surface area were a lipopolysaccharide test utilizing Limulus lysate and direct examination of the surface by scanning electron microscopy. The results obtained by the two techniques are compared and their significance to the control of microbiological slime film formation (microfouling) is discussed.     

Antibacterial copper–nickel bilayers and multilayer coatings by pulsed laser deposition on titanium

Biofouling, especially microfouling, is a major concern with the use of titanium (Ti) in the marine environment as a condenser material in cooling water systems. Earlier, copper–nickel (Cu/Ni) alloys were extensively used in marine environments due to their high corrosion and biofouling resistance. However, the choice of condenser material for the new fast breeder reactor in Kalpakkam is Ti to avoid steam side corrosion problems, which may pose a threat to steam generator parts having sodium as the secondary coolant. This study evaluates the surface modification of Ti using nano films of copper (Cu) and nickel (Ni) to utilize the antibacterial property of copper ions in reducing microfouling. The surface modification of Ti was carried out by the deposition of a Cu/Ni bilayer and (Cu/Ni)₁₀ multilayer films using a pulsed laser deposition technique. Various surface characterization studies revealed that the deposited Cu/Ni films were thin and nanocrystalline in nature. The antibacterial properties were evaluated using total viable count and epifluorescence microscopic techniques. The results showed an apparent decrease in bacterial attachment on multilayered and bilayered Cu/Ni thin films on Ti surfaces. Comparative studies between the two types of films showed a bigger reduction in numbers of microorganisms on the multilayers.     

Non-Chemical biofouling control in heat exchangers and seawater piping systems using acoustic pulses generated by an electrical discharge

Acoustic pulses generated by an electrical discharge (pulsed acoustics) were investigated as a means for biofouling control in two test formats, viz. a 5/8" outside diameter titanium tube and a mockup heat exchanger. The pulsed acoustic device, when operated at 17 kV, demonstrated 95% inhibition of microfouling over a 20 ft length of titanium tube over a 4-week period, comparable to chlorination in combination with a high-velocity flush. The pulsed acoustic device inhibited microfouling over a downstream distance of 15 ft, therefore, a single pulsed acoustic device is theoretically capable of protecting at least 30 ft of tube from microfouling (15 ft on either side of the device). A correlation between acoustic intensity in the frequency range 0.01-1 MHz and the level of biofouling inhibition was observed. The threshold acoustic intensity for microfouling inhibition was determined for this frequency range. It was also observed that the orientation of the device is critical to obtaining microfouling inhibition.     

Littoral diatoms as indicators for the eutrophication of shallow lakes

The littoral zone of shallow water bodies in the Czech Republic has been studied quite consistently at several fishponds. The use of algae, especially diatoms, for the monitoring of the state of lotic freshwater also has a long tradition. The main objective of the presented paper is to validate the feasibility of the use of littoral periphyton comunities for the biomonitoring of standing waters. At the investigated sites, littoral periphytic diatoms were studied together with selected enviromental variables (pH, conductivity, nutrients – especially total phosphorus) on three types of natural substrates (epilithon, epiphyton, epipelon). The evaluation of the diatom community was performed on the basis of the checklists of algal indicator species published by authors from the Czech Republic, Austria and the Netherlands. The data were subjected to statistical software NCCS 2000 (GLM Anova and ``Ward's minimum'' variance cluster analysis). Littoral periphytic diatoms appear to be good indicators of the fishpond water quality. The selected substrates show non-significant differences therefore the average values from all substrates were used. The best indicatory system for evaluation of Czech fishponds was van Dam's index.

     

Distribution of microorganisms lysing Pseudomonas aeruginosa and Staphylococcus aureus in the system of sewage waters

The regularities of lytic microorganisms distribution in domestic sewage have been studied. A reproduction of mesophilic gram-negative bacteria producing lytic substances against Pseudomonas aeruginosa and Staphylococcus aureus has been shown to take place at mechanical cleaning stages. In the primary sediment trap the number and the relative content of microorganisms lysing P. aeruginosa at mean temperature and the number of microorganisms lysing S. aureus are maximum. The number of gram-positive sporogenous bacteria lysing P. aeruginosa under conditions close to thermophilic does not change considerably till the secondary sediment trap and remains comparatively high. Certain stages of purification can be regarded as a source of microorganisms producing lytic substances.     

Trophic interactions in the methanogenic microbial community of low-temperature terrestrial ecosystems

The formation of methane in various ecosystems is due to the functioning of an anaerobic community, which combines trophically different groups of microorganisms. The methanogenic microbial community is a complex biological system, which responds to low temperatures by changes in its trophic structure resulting in redistributing matter flows. The enhanced activity of homoacetogenic bacteria at low temperature plays a significant role in this redistribution. Due to their relatively high growth rates and metabolic versatility, homoacetogens can successfully compete with fermenting bacteria and hydrogenotrophic methanogenic archaea for common substrates. The concentration of hydrogen is an important regulatory factor in the psychroactive methanogenic community. At low temperature methanogenic archaea possessing a higher affinity for hydrogen than homoacetogens provide for interspecies H2 transport in syntrophic reactions of fatty acid decomposition. The formation of a balanced community at low temperature is a longtime process. Cold terrestrial ecosystems are dominated by psychroactive (psychrotolerant) microorganisms, which can grow over a wide range of ambient temperatures.     

稳定性同位素探测技术在微生物生态学研究中的应用

稳定性同位素标记技术同分子生物学技术相结合而发展起来的稳定性同位素探测技术（stableisotope probing,SIP）,在对各种环境中微生物群落组成进行遗传分类学鉴定的同时,可确定其在环境过程中的功能,提供复杂群落中微生物相互作用及其代谢功能的大量信息,具有广阔的应用前景.其基本原理是：将原位或微宇宙（microcosm）的环境样品暴露于稳定性同位素富集的基质中,这些样品中存在的某些微生物能够以基质中的稳定（性同位素为碳源或氮源进行物质代谢并满足其自身生长需要,基质中的稳定性同位素被吸收同化进入微生物体内,参与各类物质如核酸（DNA和RNA）及磷脂脂肪酸（PLFA）等的生物合成,通过提取、分离、纯化、分析这些微生物体内稳定性同位素标记的生物标志物,从而将微生物的组成与其功能联系起来.在介绍稳定性同位素培养基质的选择及标记方法、合适的生物标志物的选择及提取分离方法的基础上,举例阐述了此项技术在甲基营养菌、有机污染物降解菌、根际微生物生态、互营微生物、宏基因组学等方面的应用.