Boundaries for Biofilm Formation: Humidity and Temperature

Environmental conditions which define boundaries for biofilm production could provide useful ecological information for biofilm models. A practical use of defined conditions could be applied to the high-level nuclear waste repository at Yucca Mountain. Data for temperature and humidity conditions indicate that decreases in relative humidity or increased temperature severely affect biofilm formation on three candidate canister metals.     

Correlation of extracellular polymeric substances and microbial community structure in denitrification biofilm exposed to adverse conditions

Microbial community may respond to different adverse conditions and result in the variation of extracellular polymeric substances (EPS) in denitrification biofilm; this study discovered the role of EPS in accordance with the analysis of cyclic diguanylate (c-di-GMP) and electron equilibrium (EE) under low organic loading rate, shock organic loading rate and low temperature conditions. Good nitrate removal performance could be achieved under shock organic loading rate and low temperature conditions; however, owing to the low organic loading rate, the carbon source was preferentially utilized for biomass growth. Tightly bound EPS (TB-EPS) contents progressively increased and facilitated cell adhesion and biofilm formation. The stable TB protein (TB-PN) content in TB-EPS built a cross-linked network to maintain internal biofilm structure and led to the rapid biosynthesis of polysaccharides, which could further enhance microbial adhesion and improve nitrate removal. C-di-GMP played an important role in biomass retention and biofilm formation, based on the correlation analysis of c-di-GMP and EPS. TB polysaccharide (TB-PS) contents presented a significant positive correlation with c-di-GMP content, microbial adhesion and biofilm stabilization was further enhanced through c-di-GMP regulation. In addition, a remarkable negative correlation between electron deletion rate (EDR) and TB-PN and TB-PS was discovered, and TB-PS was required to serve as energy source to enhance denitrification according to EE analysis. Surprisingly, dynamic microbial community was observed due to the drastic community succession under low temperature conditions, and the discrepancy between the dominant species for denitrification was found under shock organic loading rate and low temperature conditions. The notable increase in bacterial strains Simlicispira, Pseudomonas and Chryseobacterium was conducive to biofilm formation and denitrification under shock organic loading rate, while Dechloromonas and Zoogloea dramatically enriched for nitrate removal under low temperature conditions. The high abundance of Dechloromonas improved the secretion of EPS through the downstream signal transduction, and the c-di-GMP conserved in Pseudomonas concurrently facilitated to enhance exopolysaccharide production to shock organic loading rate and low temperature conditions.     

Diversity and biomass accumulation in cultured phototrophic biofilms

In the present study, biomass development and changes in community composition of phototrophic biofilms grown under different controlled ambient conditions (light, temperature and flow) were examined. Source communities were taken from a wastewater treatment plant and used to inoculate growth surfaces in a semi-continuous-flow microcosm. We recorded biofilm growth curves in cultures over a period of 30 days across 12 experiments. Biovolume of phototrophs and community composition for taxonomic shifts were also obtained using light and electron microscopy. Species richness in the cultured biofilms was greatly reduced with respect to the natural samples, and diversity decreased even further during biofilm development. Diadesmis confervacea, Phormidium spp., Scenedesmus spp. and Synechocystis spp. were identified as key taxa in the microcosm. While a significant positive effect of irradiance on biofilm growth could be identified, impacts of temperature and flow rate on biofilm development and diversity were less evident. We discuss the hypothesis that biofilm development could have been subject to multistability, i.e. the existence of several possible stable biofilm configurations for the same set of environmental parameters; small variations in the species composition might have been sufficient to switch between these different configurations and thus have contributed to overwriting the original effects of temperature and flow velocity.     

Colorimetric assay for biofilms in wet processing conditions

Controlling bacterial biofilms is necessary for food safety and industrial processing in clean room environments. Our goal was to develop a method to quantitatively measure biofilm produced by pathogens under wet poultry production and processing conditions. Stainless steel and glass coupons were incubated in aqueous media containing reduced nutrients and exposed to Listeria monocytogenes under static temperature and humidity conditions. Samples were measured separately by biofilm assay and viable cell density, and then confirmed by spectrophotometry and microscopy. The biofilm assay resulted in different t groupings from the cell density. The mean from the biofilm assay was 0.50, and the error% was 0.595. The mean of the log₁₀ density (cfu/cm²) was 5.90, and the standard deviation ranged from 0.127 to 0.438 on 24 coupons. The typical sequence of biofilm development, followed by microscopy of biofilm grown on glass coupons, exhibited a change from dispersed single cells to an all-over pattern of clumps with few dispersed cells. L. monocytogenes formed biofilms on all of the substrata tested. Bacterial counts from planktonic cultures at 24, 48, 72, and 144 h confirmed that L. monocytogenes remained viable throughout the experiment and reached equilibrium between 6 and 24 h. The cell density log₁₀/ml was 8.01, 8.03, 7.69, and 6.66, respectively; and the standard deviation ranged from 0.156 to 0.394. The data will be used to grow stable biofilms of Listeria spp. collected from the food processing environment for further study. This is the first use of the crystal violet assay for measurement of bacterial biofilms on stainless steel under these conditions. The methods tested are applicable to other bacteria and substrata.     

The role of microbial biofilms in deterioration of space station candidate materials

Formation of microbial biofilms on surfaces of a wide range of materials being considered as candidates for use on the International Space Station was investigated. The materials included a fibre-reinforced polymeric composite, an adhesive sealant, a polyimide insulation foam, teflon cable insulation, titanium, and an aliphatic polyurethane coating. They were exposed to a natural mixed population of bacteria under controlled conditions of temperature and relative humidity (RH). Biofilms formed on the surfaces of the materials at a wide range of temperatures and RHs. The biofilm population was dominated by Pseudomonas aeruginosa, Ochrobactrum anthropi, Alcaligenes denitrificans, Xanthomonas maltophila, and Vibrio harveyi. The biocide, diiodomethyl-p-tolyl sulfone, impregnated in the polyurethane coating, was ineffective against microbial colonization and growth. Degradation of the polyurethane coatings was monitored with electrochemical impedance spectroscopy (EIS). The impedance spectra indicated that microbial degradation of the coating occurred in several stages. The initial decreases in impedance were due to the transport of water and solutes into the polymeric matrices. Further decreases were a result of polymer degradation by microorganisms. Our data showed that these candidate materials for space application are susceptible to biofilm formation and subsequent degradation. Our study suggests that candidate materials for use in space missions need to be carefully evaluated for their susceptibility to microbial biofilm formation and biodegradation.     

一株工业腐败物中魏氏柠檬酸杆菌的分离鉴定及产生物膜特性分析

【目的】分离和鉴定工业腐败物中高产细菌生物膜菌株,并明确该菌的部分产膜特性。【方法】通过微孔板结晶紫染色法对分离的菌株进行产膜能力评价,根据菌落形态、生理生化特性和16S rRNA序列的系统进化树分析进行菌株鉴定;同时利用扫描电子显微镜(SEM)和结晶紫染色法分别研究材料及温度对该菌产膜特性和能力的影响。【结果】筛选出一株高产细菌生物膜菌株,经鉴定该菌为魏氏柠檬酸杆菌;其在玻璃、不锈钢和聚氯乙烯(PVC)材料表面均能形成生物膜;温度条件显著影响产膜能力,在30°C时,菌株在PVC材料表面形成生物膜能力最强。【结论】工业腐败物中含有高产细菌生物膜菌株,并且产膜受附着物和温度影响。     

Oxidative and nitrosative stress in Staphylococcus aureus biofilm

Diverse chemical and physical agents can alter cellular functions associated with oxidative metabolism, thus stimulating the production of reactive oxygen species (ROS) and reactive nitrogen intermediates (RNI) in planktonic bacterial physiology. However, more research is necessary to determine the precise role of cellular stress in biofilm. The present study was designed to address the issues of Staphylococcus aureus biofilm formation with respect to the generation of oxidative and nitrosative stress. We studied three pathogenic S. aureus clinical strains and an ATCC strain exposed to a different range of culture conditions (time, temperature, pH, reduction and atmospheric conditions) using quantitative methods of biofilm detection. We observed that cellular stress could be produced inside biofilms, thereby affecting their growth, resulting in an increase of ROS and RNI production, and a decrease of the extracellular matrix under unfavorable conditions. These radical oxidizers could then accumulate in an extracellular medium and thus affect the matrix. These results contribute to a better understanding of the processes that enable adherent biofilms to grow on inert surfaces and lead to an improved knowledge of ROS and RNI regulation, which may help to clarify the relevance of biofilm formation in medical devices.     

温湿度对稻纵卷叶螟卵的联合作用

为了探讨温湿度在稻纵卷叶螟Cnaphalocrocis medinalis （Guenée）种群发展中的作用, 通过室内实验调查了不同温度和湿度组合下该蛾卵的发育历期、 胚胎发育情况、 孵化率和卵粒重量的变化。结果表明： 在相同温度下卵历期随相对湿度的增大而缩短, 孵化率随相对湿度的加大而提高。在22℃下低于46%的相对湿度显著降低了卵的孵化率, 而在25～34℃下低于66%的相对湿度会引起孵化率的显著降低, 37℃下卵无论在何种湿度中均不能孵化。在50%左右的低湿条件下, 温度高于28℃后卵也不能孵化。温度在22～31℃和相对湿度在77%～100%范围内, 卵的孵化率无显著差异, 这属于稻纵卷叶螟卵的适宜温湿度范围。稻纵卷叶螟卵的发育起点温度和有效积温分别为10.1±0.6℃和63.7±3.5日度。卵的孵化率（Y）与温湿系数（RH/T）间呈显著的逻辑斯蒂曲线关系Y=0.8662/[1+exp(17.4084-7.5714×RH/T)]。温湿系数在2.34以下时卵孵化率将低于50%, 而达到3.0左右时孵化率接近最高值。结论认为, 低湿造成的稻纵卷叶螟卵重量显著降低、 卵粒干瘪、 胚胎发育受阻是致死卵的主要原因。     

Biofilm Formation by the Fish Pathogen Flavobacterium columnare: Development and Parameters Affecting Surface Attachment

Flavobacterium columnare is a bacterial fish pathogen that affects many freshwater species worldwide. The natural reservoir of this pathogen is unknown, but its resilience in closed aquaculture systems posits biofilm as the source of contagion for farmed fish. The objectives of this study were (i) to characterize the dynamics of biofilm formation and morphology under static and flow conditions and (ii) to evaluate the effects of temperature, pH, salinity, hardness, and carbohydrates on biofilm formation. Nineteen F. columnare strains, including representatives of all of the defined genetic groups (genomovars), were compared in this study. The structure of biofilm was characterized by light microscopy, confocal laser scanning microscopy, and scanning electron microscopy. F. columnare was able to attach to and colonize inert surfaces by producing biofilm. Surface colonization started within 6 h postinoculation, and microcolonies were observed within 24 h. Extracellular polysaccharide substances and water channels were observed in mature biofilms (24 to 48 h). A similar time course was observed when F. columnare formed biofilm in microfluidic chambers under flow conditions. The virulence potential of biofilm was confirmed by cutaneous inoculation of channel catfish fingerlings with mature biofilm. Several physicochemical parameters modulate attachment to surfaces, with the largest influence being exerted by hardness, salinity, and the presence of mannose. Maintenance of hardness and salinity values within certain ranges could prevent biofilm formation by F. columnare in aquaculture systems.     

Poly(3-hydroxybutyrate) influences biofilm formation and motility in the novel Antarctic species <Emphasis Type="Italic">Pseudomonas extremaustralis</Emphasis> under cold conditions

Polyhydroxyalkanoates (PHAs) are highly reduced bacterial storage compounds that increase fitness in changing environments. It has previously shown that polyhydroxybutyrate (PHB) accumulation is essential during the growth under cold conditions. In this work, the relationship between PHB accumulation and biofilm development at low temperature was investigated. P. extremaustralis, an Antarctic strain able to accumulate PHB, and its phaC mutant, impaired in the synthesis of this polymer, were used to analyze microaerobic growth, biofilm development, EPS content and motility. PHB accumulation increased motility and survival of planktonic cells in the biofilms developed by P. extremaustralis under cold conditions. Microaerobic conditions rescued the cold growth defect of the mutant strain. The PHB accumulation capability could constitute an adaptative advantage for the colonization of new ecological niches in stressful environments.     

Response of Tomato Sugar and Acid Metabolism and Fruit Quality under Different High Temperature and Relative Humidity Conditions

The combined stress of high temperature and high relative air humidity is one of the most serious agrometeorological disasters that restricts the production capacity of protected agriculture. However, there is little information about the precise interaction between them on tomato fruit quality. The objectives of this study were to explore the effects of the combined stress of high temperature and relative humidity on the sugar and acid metabolism and fruit quality of tomato fruits, and to determine the best relative air humidity for fruit quality under high temperature environments. Four temperature treatments (32°C, 35°C, 38°C, 41°C), three relative air humidity (50%, 70%, 90%) and four duration (3, 6, 9, 12 d) orthogonal experiments were conducted, with 28°C, 50% as control. The results showed that under high temperature and relative air humidity, the activity of sucrose metabolizing enzymes in young tomato fruits changed, which reduced fruits soluble sugar content; in addition, enzyme activities involved phosphopyruvate carboxylase (PEPC), mitochondria aconitase (MDH) and citrate synthetase (CS) increased which increased the content of organic acids (especially malic acid). Eventually, vitamin C, total sugar and sugar-acid ratio decreased significantly, while the titratable acid increased, resulting in a decrease in fruit flavor quality and nutritional quality in ripe fruit. Specifically, a temperature of 32°C and a relative air humidity of 70% were the best cultivation conditions for tomato reproductive growth period under high temperature. Our results indicating that fruit quality reduced under high temperature at the flowering stage, while increasing the relative air humidity to 70% could alleviate this negative effect. Our results are benefit to better understand the interaction between microclimate parameters under specific climatic conditions in the greenhouse environment and their impact on tomato flavor quality.     

Biofilm formation and sloughing in Serratia marcescens are controlled by quorum sensing and nutrient cues

We describe here a role for quorum sensing in the detachment, or sloughing, of Serratia marcescens filamentous biofilms, and we show that nutrient conditions affect the biofilm morphotype. Under reduced carbon or nitrogen conditions, S. marcescens formed a classical biofilm consisting of microcolonies. The filamentous biofilm could be converted to a microcolony-type biofilm by switching the medium after establishment of the biofilm. Similarly, when initially grown as a microcolony biofilm, S. marcescens could be converted back to a filamentous biofilm by increasing the nutrient composition. Under high-nutrient conditions, an N-acyl homoserine lactone quorum-sensing mutant formed biofilms that were indistinguishable from the wild-type biofilms. Similarly, other quorum-sensing-dependent behaviors, such as swarming motility, could be rendered quorum sensing independent by manipulating the growth medium. Quorum sensing was also found to be involved in the sloughing of the filamentous biofilm. The biofilm formed by the bacterium consistently sloughed from the substratum after approximately 75 to 80 h of development. The quorum-sensing mutant, when supplemented with exogenous signal, formed a wild-type filamentous biofilm and sloughed at the same time as the wild type, and this was independent of surfactant production. When we removed the signal from the quorum-sensing mutant prior to the time of sloughing, the biofilm did not undergo significant detachment. Together, the data suggest that biofilm formation by S. marcescens is a dynamic process that is controlled by both nutrient cues and the quorum-sensing system.     

The Evaporative Function of Cockroach Hygroreceptors

Insect hygroreceptors associate as antagonistic pairs of a moist cell and a dry cell together with a cold cell in small cuticular sensilla on the antennae. The mechanisms by which the atmospheric humidity stimulates the hygroreceptive cells remain elusive. Three models for humidity transduction have been proposed in which hygroreceptors operate either as mechanical hygrometers, evaporation detectors or psychrometers. Mechanical hygrometers are assumed to respond to the relative humidity, evaporation detectors to the saturation deficit and psychrometers to the temperature depression (the difference between wet-bulb and dry-bulb temperatures). The models refer to different ways of expressing humidity. This also means, however, that at different temperatures these different types of hygroreceptors indicate very different humidity conditions. The present study tested the adequacy of the three models on the cockroach’s moist and dry cells by determining whether the specific predictions about the temperature-dependence of the humidity responses are indeed observed. While in previous studies stimulation consisted of rapid step-like humidity changes, here we changed humidity slowly and continuously up and down in a sinusoidal fashion. The low rates of change made it possible to measure instantaneous humidity values based on UV-absorption and to assign these values to the hygroreceptive sensillum. The moist cell fitted neither the mechanical hygrometer nor the evaporation detector model: the temperature dependence of its humidity responses could not be attributed to relative humidity or to saturation deficit, respectively. The psychrometer model, however, was verified by the close relationships of the moist cell’s response with the wet-bulb temperature and the dry cell’s response with the dry-bulb temperature. Thus, the hygroreceptors respond to evaporation and the resulting cooling due to the wetness or dryness of the air. The drier the ambient air (absolutely) and the higher the temperature, the greater the evaporative temperature depression and the power to desiccate.     

水热环境对黑眶蟾蜍幼体存活的影响

禁食后在9种水热条件下,观测了不同温湿度对黑眶蟾蜍(Bufo melanostictus Schneider)幼体存活时间和动用体内贮能情况的影响。黑眶蟾蜍幼体在24℃浅水环境中存活时间长达22d。在高温和干燥条件下幼体的存活时间最短,32℃干燥环境中幼体存活时间不到1d。不同温湿度环境下幼体体内含脂量有显著差异。在高温和干燥环境中幼体存活时间短,动用贮能少,其体内脂肪含量和总能量动用少于其它温湿度环境中的幼体。结果表明,黑眶蟾蜍幼体适宜生活在24℃浅水和湿润环境中,觅食不足并非影响其幼体存活时间的主要因子,而高温和干燥是影响其幼体存活时间的关键因子。     

Relative humidity is a key factor in the acclimation of the stomatal response to CO(2)

Previous work has shown that stomata of growth chamber-grown Vicia faba leaves have an enhanced CO2 response when compared with stomata of greenhouse-grown plants. This guard cell response to CO2 acclimatizes to the environmental conditions on the transfer of plants between the two environments. In the present study, air relative humidity is identified as a key environmental factor mediating the changes in stomatal sensitivity to CO2. In the greenhouse environment, elevation of relative humidity to growth chamber levels resulted in an enhanced CO2 response, whereas a reduction in the light level to that comparable to growth chamber conditions had no effect on stomatal CO2 sensitivity. The transfer of plants between humidified and normal greenhouse conditions resulted in an acclimation response with a time-course matching that previously obtained in transfers of plants between greenhouse and growth chamber environments. The high stomatal sensitivity to CO2 of growth chamber-grown plants could be reduced by lowering growth chamber relative humidity and then restored with its characteristic acclimation time-course by an elevation of relative humidity. Leaf temperature was unchanged during this restoration, eliminating it as a primary factor in the acclimation response. Humidity regulation of stomatal CO2 sensitivity could function as a signal for leaves inside dense foliage canopies, promoting stomatal opening under low light, low CO2 conditions.     

Labile and Recalcitrant Organic Matter Utilization by River Biofilm Under Increasing Water Temperature

Microbial biofilms in rivers contribute to the decomposition of the available organic matter which typically shows changes in composition and bioavailability due to their origin, seasonality, and watershed characteristics. In the context of global warming, enhanced biofilm organic matter decomposition would be expected but this effect could be specific when either a labile or a recalcitrant organic matter source would be available. A laboratory experiment was performed to mimic the effect of the predicted increase in river water temperature (+4?°C above an ambient temperature) on the microbial biofilm under differential organic matter sources. The biofilm microbial community responded to higher water temperature by increasing bacterial cell number, respiratory activity (electron transport system) and microbial extracellular enzymes (extracellular enzyme activity). At higher temperature, the phenol oxidase enzyme explained a large fraction of respiratory activity variation suggesting an enhanced microbial use of degradation products from humic substances. The decomposition of hemicellulose (β-xylosidase activity) seemed to be also favored by warmer conditions. However, at ambient temperature, the enzymes highly responsible for respiration activity variation were β-glucosidase and leu-aminopeptidase, suggesting an enhanced microbial use of polysaccharides and peptides degradation products. The addition of labile dissolved organic carbon (DOC; dipeptide plus cellobiose) caused a further augmentation of heterotrophic biomass and respiratory activity. The changes in the fluorescence index and the ratio Abs(250)/total DOC indicated that higher temperature accelerated the rates of DOC degradation. The experiment showed that the more bioavailable organic matter was rapidly cycled irrespective of higher temperature while degradation of recalcitrant substances was enhanced by warming. Thus, pulses of carbon at higher water temperature might have consequences for DOC processing.     

Diurnal flight behavior of Ichneumonoidea (Insecta: Hymenoptera) related to environmental factors in a tropical dry forest

The biology and behavior of insects are strongly influenced by environmental conditions such as temperature and precipitation. Because some of these factors present a within day variation, they may be causing variations on insect diurnal flight activity, but scant information exists on the issue. The aim of this work was to describe the patterns on diurnal variation of the abundance of Ichneumonoidea and their relation with relative humidity, temperature, light intensity, and wind speed. The study site was a tropical dry forest at Ría Lagartos Biosphere Reserve, Mexico; where correlations between environmental factors (relative humidity, temperature, light, and wind speed) and abundance of Ichneumonidae and Braconidae (Hymenoptera: Ichneumonoidea) were estimated. The best regression model for explaining abundance variation was selected using the second order Akaike Information Criterion. The optimum values of temperature, humidity, and light for flight activity of both families were also estimated. Ichneumonid and braconid abundances were significantly correlated to relative humidity, temperature, and light intensity; ichneumonid also showed significant correlations to wind speed. The second order Akaike Information Criterion suggests that in tropical dry conditions, relative humidity is more important that temperature for Ichneumonoidea diurnal activity. Ichneumonid wasps selected toward intermediate values of relative humidity, temperature and the lowest wind speeds; while Braconidae selected for low values of relative humidity. For light intensity, braconids presented a positive selection for moderately high values.     

一株吡啶降解菌Pseudomonas sp.ZX08的生物膜形成特性及影响因素

【背景】煤化工企业排放的废水中含有大量难降解、高毒性的有机污染物,采用以高效降解菌为基础的生物强化技术对其进行处理,是一种经济可行的策略;而促进高效降解菌在载体材料表面的生物膜形成,有助于提升生物膜法废水处理系统的效能。【目的】探究一株吡啶高效降解菌Pseudomonas sp. ZX08的生物膜形成过程和特性,识别不同的环境因子如温度、pH、Na⁺、K⁺、Ca²⁺、Mg²⁺等对其生物膜形成的影响规律,为实现人工调控其在实际废水处理系统中的成膜过程提供理论依据。【方法】采用改良的微孔板生物膜培养与定量方法,以单因子影响实验测定不同条件下菌株在12孔板内的生物膜形成量和浮游态细菌量;采用激光共聚焦显微镜(confocallaserscanning microscope,CLSM)观察和分析生物膜的结构特征。【结果】Pseudomonas sp. ZX08菌株具有良好的吡啶降解性能,且生物膜形成能力较强,CLSM观察到其在载体表面形成的生物膜可达40-50mm;生物膜外层的活细胞比例更高,分泌的胞外蛋白...     

Biofilm formation at warming temperature: acceleration of microbial colonization and microbial interactive effects

River biofilms that grow on wet benthic surface are mainly composed of bacteria, algae, cyanobacteria and protozoa embedded in a polysaccharide matrix. The effects of increased river water temperature on biofilm formation were investigated. A laboratory experiment was designed employing two temperatures (11.1-13.2°C, night-day; 14.7-16.0°C, night-day) and two nutrient levels (0.054 mg P l(-1), 0.75 mg N l(-1); 0.54 mg P l(-1), 7.5 mg N l(-1)). Biofilm formation at the higher temperature was faster, while the biomass of the mature biofilm was mainly determined by nutrient availability. The specific response of the three microbial groups that colonized the substrata (algae, bacteria and ciliates) was modulated by interactions between them. The greater bacterial growth rate and earlier bacterial colonization at the higher temperature and higher nutrient status was not translated into the accrual of higher bacterial biomass. This may result from ciliates grazing on the bacteria, as shown by an earlier increase in peritrichia at higher temperatures, and especially at high nutrient conditions. Temperature and ciliate grazing might determine the growth of a distinctive bacterial community under warming conditions. Warmer conditions also produced a thicker biofilm, while functional responses were much less evident (increases in the heterotrophic utilization of polysaccharides and peptides, but no increase in primary production and respiration). Increasing the temperature of river water might lead to faster biofilm recolonization after disturbances, with a distinct biofilm community structure that might affect the trophic web. Warming effects would be expected to be more relevant under eutrophic conditions.     

Effect of Batch and Fed-Batch Growth Modes on Biofilm Formation by <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> at Different Temperatures

The influence of Listeria monocytogenes (L. monocytogenes) biofilm formation feeding conditions (batch and fed-batch) at different temperatures on biofilm biomass and activity was determined. Biofilm biomass and cellular metabolic activity were assessed by Crystal Violet (CV) staining and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (XTT) colorimetric method, respectively. Live/Dead staining was also performed in order to get microscopic visualization of the different biofilms. Results revealed that at refrigeration temperature (4°C) a higher amount of biofilm was produced when batch conditions were applied, while at higher temperatures the fed-batch feeding condition was the most effective on biofilm formation. Moreover, independently of the temperature used, biofilms formed under fed-batch conditions were metabolically more active than those formed in batch mode. In conclusion, this work shows that different growth modes significantly influence L. monocytogenes biofilm formation on abiotic surfaces as well as the metabolic activity of cells within biofilms.