Survival Of Bacterial Cells Research Articles

Drying kinetics and survival studies of dairy fermentation bacteria in convective air drying environment using single droplet drying

The drying and survival kinetics of Lactococcus lactis ssp. cremoris in a convective air drying environment were measured using single droplet drying experiments. Tests were carried out at five different drying temperatures (45–95°C) at a constant air velocity (0.5m/s) and within 2.4–11% relative humidity. The effect of protective agents (10% w/w) of lactose, sodium caseinate and lactose:sodium caseinate (3:1) was also evaluated. The thermal inactivation kinetics parameters in convective air drying and isothermal water bath heating were determined and compared. The results showed that the final temperature attained by the droplet affected the survival of the bacteria significantly, however, most of the bacterial death occurred in early stage of drying while evaporative cooling kept the drop temperature relatively low. At higher droplet temperatures (⩾65°C) the bacterial cultures were inactivated by both dehydration and thermal stresses. At lower droplet temperatures (⩽55°C) the rate of change in droplet moisture content had much stronger effect on the bacterial survival. Lactose and sodium caseinate, as protective agents, enhanced the survival of bacterial cells significantly at all the test conditions. The lactose:sodium caseinate (3:1) mixture synergistically enhanced the survival of the bacterial cultures. The death of these bacteria followed first-order kinetics during convective single droplet drying as well as during isothermal water-bath heating. However, the inactivation energy in convective single droplet drying (181.3kJ/mol) was much higher than the inactivation energy in isothermal water bath heating (16.8kJ/mol) within the medium temperature of 45–95°C.

Exploring the antibacterial properties of honey and its potential

Exploring the antibacterial properties of honey and its potential

Optimization of operating conditions for the application of Moringa oleifera (Zogale) seeds extract in water disinfection using response surface methodology

Good quality dry seeds of Moringa oleifera were selected and the seed coat and wings were removed manually. The kernel was ground to fine powder using the coffee mill attachment of the Moulinex domestic food blender. The ground powder was then sieved through 210 im sieve. The seed powder was de-fatted using hexane in electro-thermal Soxhlet extractor. Moringa seeds extract was obtained using the de-fatted seed cake and aqueous extraction. Different preparations of Moringa extract were added to 10 mL of the Escherichia coli suspension and incubated for 2 h without agitation. Survival of bacterial cells was assessed by making dilution series of bacterial suspensions, plating on non-selective LB medium agar dishes, and incubating for 48 h at 37°C. Duplicates were made of every individual assay. Colonies were counted on dishes and the bacterial cell survival ratio was estimated by comparison to a control experiment where no Moring extract was added. The bacterial removal was optimized by varying the mixing time, mixing speed, and Moringa seeds extract dosage. Statistical optimization was conducted by using central composite design (CCD). The experimental data was analyzed using statistical software DESIGN EXPERT, V6.4.8 DEC 10 2002 for Windows. The response surface model was used to determine the optimum operating condition that yields the highest antimicrobial compounds activities from M. oleifera seeds extracts. A cubic model was fitted to the data. The standard deviation for the cubic model was 0.56, with R 2 = 0.9999 and adjusted R 2 = 0.9994. The analysis of variance (ANOVA) showed that the effects of mixing time, mixing speed and Moringa dosage were significant (p < 0.05) in the extraction process. The Quadratic model was used in predicting the responses and the optimal conditions were determined as 31 min mixing time, 85 rpm mixing speed and 3.25 mg/mL Moringa dosage. The results show that the predicted and experimental values were not significantly different and it was thus concluded that the model obtained can be used to optimize the process of antimicrobial bioactive compound extraction from de-fatted M. oleifera seeds. Key words: Moringa oleifera, seeds extract, bacterial inactivation, optimization, response surface model.

Stability of spore-based biosensing systems under extreme conditions

The full exploitation of bacterial whole-cell biosensing systems in field applications requires the survival of bacterial cells and long-term preservation of their sensing ability during transportation and on-site storage of such analytical systems. Specifically, there is a need for rapid, simple and inexpensive biosensing systems for monitoring human health and the environment in remote areas as well as developing countries, which often suffer from harsh atmospheric conditions and inadequate commercial distribution and storage facilities. Our laboratory has previously reported the successful use of bacterial spores as vehicles for the long-term preservation and storage of whole-cell biosensing systems at room temperature. In the present research, we have accomplished a year-long study to investigate the effect of extreme climatic conditions on the stability of spores-based whole-cell biosensing systems. The spores were stored in laboratory conditions that simulated those found in real harsh environments, including, extreme temperatures and humidity levels, and desiccation. This study was crucial in determining the germination ability and analytical performance of the spore-based sensing systems upon storage in such conditions in order to support their effective use in the field, in extreme environments. Our results proved that the intrinsic resistance of spores to harsh environmental conditions helped maintain the integrity of the encapsulated sensor bacteria. The revived active cells actually retained their analytical performance during the course of the 12-month storage study. We envision that spore-based sensing systems could pave the way to the use of whole-cell biosensors in field applications and in areas where they have not been employed so far.

Impact of improved potassium accumulation on pH homeostasis, membrane potential adjustment and survival of Corynebacterium glutamicum

Metal ion uptake is crucial for all living cells and an essential part of cellular bioenergetic homeostasis. In this study the uptake and the impact of the most abundant internal cation, potassium, were investigated in Actinobacteria, a group of high G + C Gram-positives with a number of prominent biotechnologically and medically important members. Genome analyses revealed a variety of different potassium uptake systems in this monophyletic group ranging from potassium channels common in virtually all Actinobacteria to different active carriers that were present predominantly in pathogenic members able to cope with various stress conditions. By applying Corynebacterium glutamicum as model system we provide experimental evidence that under optimal conditions a potassium channel is sufficient in bacteria for the maintenance of internal pH and membrane potential ensuring survival of cells under stress conditions. Under potassium limitation, however, viability of C. glutamicum was increased under acidic stress or during desiccation when a functional KtrAB potassium transporter from the pathogen Corynebacterium jeikeium was heterologously expressed. We provide experimental evidence that the KtrAB mediated enhanced potassium accumulation improved maintenance of internal pH and membrane potential. The results indicate that the occurrence of active potassium transport systems correlates with an improved potassium-dependent bioenergetic homeostasis and survival of bacterial cells under stress conditions.

Re-examining the role of hydrogen peroxide in bacteriostatic and bactericidal activities of honey.

The aim of this study was to critically analyze the effects of hydrogen peroxide on growth and survival of bacterial cells in order to prove or disprove its purported role as a main component responsible for the antibacterial activity of honey. Using the sensitive peroxide/peroxidase assay, broth microdilution assay and DNA degradation assays, the quantitative relationships between the content of H2O2 and honey’s antibacterial activity was established. The results showed that: (A) the average H2O2 content in honey was over 900-fold lower than that observed in disinfectants that kills bacteria on contact. (B) A supplementation of bacterial cultures with H2O2 inhibited E. coli and B. subtilis growth in a concentration-dependent manner, with minimal inhibitory concentrations (MIC90) values of 1.25 mM/107 cfu/ml and 2.5 mM/107 cfu/ml for E. coli and B. subtilis, respectively. In contrast, the MIC90 of honey against E. coli correlated with honey H2O2 content of 2.5 mM, and growth inhibition of B. subtilis by honey did not correlate with honey H2O2 levels at all. (C) A supplementation of bacterial cultures with H2O2 caused a concentration-dependent degradation of bacterial DNA, with the minimum DNA degrading concentration occurring at 2.5 mM H2O2. DNA degradation by honey occurred at lower than ≤2.5 mM concentration of honey H2O2 suggested an enhancing effect of other honey components. (D) Honeys with low H2O2 content were unable to cleave DNA but the addition of H2O2 restored this activity. The DNase-like activity was heat-resistant but catalase-sensitive indicating that H2O2 participated in the oxidative DNA damage. We concluded that the honey H2O2 was involved in oxidative damage causing bacterial growth inhibition and DNA degradation, but these effects were modulated by other honey components.

Burkholderia Type VI Secretion Systems Have Distinct Roles in Eukaryotic and Bacterial Cell Interactions

Bacteria that live in the environment have evolved pathways specialized to defend against eukaryotic organisms or other bacteria. In this manuscript, we systematically examined the role of the five type VI secretion systems (T6SSs) of Burkholderia thailandensis (B. thai) in eukaryotic and bacterial cell interactions. Consistent with phylogenetic analyses comparing the distribution of the B. thai T6SSs with well-characterized bacterial and eukaryotic cell-targeting T6SSs, we found that T6SS-5 plays a critical role in the virulence of the organism in a murine melioidosis model, while a strain lacking the other four T6SSs remained as virulent as the wild-type. The function of T6SS-5 appeared to be specialized to the host and not related to an in vivo growth defect, as ΔT6SS-5 was fully virulent in mice lacking MyD88. Next we probed the role of the five systems in interbacterial interactions. From a group of 31 diverse bacteria, we identified several organisms that competed less effectively against wild-type B. thai than a strain lacking T6SS-1 function. Inactivation of T6SS-1 renders B. thai greatly more susceptible to cell contact-induced stasis by Pseudomonas putida, Pseudomonas fluorescens and Serratia proteamaculans—leaving it 100- to 1000-fold less fit than the wild-type in competition experiments with these organisms. Flow cell biofilm assays showed that T6S-dependent interbacterial interactions are likely relevant in the environment. B. thai cells lacking T6SS-1 were rapidly displaced in mixed biofilms with P. putida, whereas wild-type cells persisted and overran the competitor. Our data show that T6SSs within a single organism can have distinct functions in eukaryotic versus bacterial cell interactions. These systems are likely to be a decisive factor in the survival of bacterial cells of one species in intimate association with those of another, such as in polymicrobial communities present both in the environment and in many infections.

Halophilic Archaea cultured from ancient halite, Death Valley, California

Halophilic Archaea cultured from ancient fluid inclusions in a 90-m-long (0- to 100,000-year-old) salt core from Death Valley, California, demonstrate survival of bacterial cells in subsurface halite for up to 34,000 years. Five enrichment cultures, representing three genera of halophilic Archaea (Halorubrum, Natronomonas and Haloterrigena), were obtained from five surface-sterilized halite crystals exclusively in one section of the core (13.0-17.8 m; 22,000-34,000 years old) containing perennial saline lake deposits. Prokaryote cells were observed microscopically in situ within fluid inclusions from every layer that produced culturable cells. Another 876 crystals analysed from depths of 8.1-86.7 m (10,000-100,000 years old) failed to yield live halophilic Archaea. Considering the number of halite crystals tested (culturing success of 0.6%), microbial survival in fluid inclusions in halite is rare and related to the paleoenvironment, which controls the distribution and abundance of trapped microorganisms. Two cultures from two crystals at 17.8 m that yielded identical 16S rRNA sequences (genus: Haloterrigena) demonstrate intra-laboratory reproducibility. Inter-laboratory reproducibility is shown by two halophilic Archaea (genus: Natronomonas), with 99.3% similarity of 16S rRNA sequences, cultured from the same core interval, but at separate laboratories.

Site‐directed mutagenesis of rat thioltransferase: Effects of essential cysteine residues for the protection against oxidative stress

A cDNA of rat liver thioltransferase was cloned and then expressed using pMAL-c expression vector in Escherichia coli. Recombinant rat liver thioltransferase was expressed as a fusion protein with maltose-binding protein and then purified by amylose resin column chromatography to be homogeneity on 12.5% SDS-polyacrylamide gel electrophoretic analysis. The expressed proteins were shown as two bands at around 53 and 41 kDa, suggesting that the high molecular one was a fusion protein of recombinant thioltransferase (11.7 plus 41 kDa) and the other (smaller one) was a maltose-binding protein (41 kDa). A recombinant thioltransferase catalyzed a thiol/disulfide exchange reaction in the same way as thioltransferases purified from various sources. Compared with wild type, the mutants C23A, C26A, C79A, and C83A showed 0%, 17%, 82%, and 86% in the enzymatic activity, respectively. In addition, wild-type-transfected bacteria expressed in bacterial cells showed a strong resistance to H(2)O(2) treatment as well as the case of active mutants (C79A and C83A), but inactive mutants (C23A and C26A) showed no resistance to H(2)O(2) treatment as same as mocktransfection. Thioltransferase can be important for survival of bacterial cells under oxidative stress.

Targeting the Peptide Deformylase of Mycobacterium tuberculosis Leads to Drug Discovery

Abstract: Peptide deformylase (PDF) is a ubiquitous bacterial metalloenzyme responsible to cleave the formyl group from nascent polypeptides, supporting in the maturation. It plays a vital role in the survival of bacterial cells which is con-served in the eubacteria and is considered to be an attractive target for developing new antibacterial agents. Homology modeling was employed for generation of 3-D structure of PDF of M. tuberculosis H37Rv and showed 92.5% amino acid in the allowed region of Ramachandran plot. PDF was used as target for a set of inhibitors with substantial structural dif-ferences. Docking results show that the BB-3497, BBS-54, Actinonin and BBS-02 bind with high affinity to enzyme ac-tive site. Phylogeny of PDF in M. tuberculosis H37Rv shows homology with other strains of pathogenic bacteria. These data validate PDF as a novel target for the design of a new generation of antimycobacterial agents. Keywords: Mycobacterium tuberculosis, Peptide deformylase, Docking, Phylogeny, Drugs.

A possible mechanism for visible light‐induced wound healing

Chronic wounds resistant to conventional therapy have been treated successfully with low energy lasers and light emitting diodes (LEDs) in the visible and near IR region. It has been proposed that production of low level reactive oxygen species (ROS) following illumination is the first step of photobiomodulation. It was also shown that white light (400-800 nm) has similar stimulatory effects as lasers and LEDs. ROS at higher levels are toxic to cells and bacteria. In the present study, we examined the phototoxicity of broadband (400-800 nm, 120 J/cm(2)) visible light on the survival of several pathogenic bacteria: Staphylococcus aureus 195, Pseudomonas aeruginosa 1316, Escherichia coli 1313, and Serratia marcescens. These bacteria were chosen due to their high prevalence in infected wounds. The survival of bacterial cells following illumination was monitored by counting the number of colony forming units before and after exposure to light. Illumination with white light, 120 J/cm(2), caused a reduction of 62%, 83%, and 56% in the colony count of E. coli 1313, S. aureus 195 and S. marcescens, respectively, though no reduction in the viability of P. aeruginosa 1316 was demonstrated. The phototoxic effect was found to involve induction of ROS production by the bacteria. It was also found that illumination of S. aureus 195 and E. coli 1313 in the presence of pyocyanin, known to be secreted by P. aeruginosa, had a stronger bactericidal effect compared to illumination alone. Visible light at high intensity can kill bacteria in infected wounds. Thus, illumination of infected wounds with intense visible light, prior to low intensity illumination for stimulating wound closure, may reduce infection and promote healing.

Mutations in two global regulators lower individual mortality in Escherichia coli

There has been considerable investigation into the survival of bacterial cells under stress conditions, but little is known about the causes of mortality in the absence of exogenous stress. That there is a basal frequency of cell death in such populations may reflect that it is either impossible to avoid all lethal events, or alternatively, that it is too costly. Here, through a genetic screen in the model organism Escherichia coli, we identify two mutants with lower frequencies of mortality: rssB and fliA. Intriguingly, these two genes both affect the levels of different sigma factors within the cell. The rssB mutant displays enhanced resistance to multiple external stresses, possibly indicating that the cell gains its increased vitality through elevated resistance to spontaneous, endogenous stresses. The loss of fliA does not result in elevated stress resistance; rather, its survival is apparently due to a decreased physical stress linked to the insertion of the flagellum through the membrane and energy saved through the loss of the motor proteins. The identification of these two mutants implies that reducing mortality is not impossible; rather, due to its cost, it is subject to trade-offs with other traits that contribute to the competitive success of the organism.

Effect of different environmental conditions on the bacteria survival on stainless steel surfaces

The effect of different environmental conditions on the survival of Enterobacter cloacae, Pseudomonas aeruginosa and Staphylococcus aureus on stainless steel surfaces was determined using direct epifluorescence microscopy (DEM) and plate count method. A strong linear relation ( R 2 = 0.97; P < 0.01) between DEM and plate counts over the range of 10 4–10 7 CFU/cm 2 was observed when counts by DEM were performed over viable microorganisms. However, when microorganisms were subjected to adverse conditions, such as slow air-drying, the correlation between both methods diminished considerably ( R 2 = 0.225). This may be due to microorganisms subjected to adverse conditions can be injured, so some bacteria can not be detected by cultivation, but they still present a danger. Microorganisms on wet surfaces had the ability to produce biofilm after 24 h. However, air-drying on surfaces, reduced considerably the bacterial count. E. cloacae was the most susceptible to slow air-drying on surfaces at room temperature however, S. aureus, remained viable for at least 72 h. The characteristics of an organism and its surrounding environment are among the important factors that may affect the survival of bacterial cells on surfaces.

Larval settlement and metamorphosis of the mussel Mytilus galloprovincialis in response to biofilms

Biofilms were allowed to develop on glass slips immersed 1.0–1.5 m below the sea surface in Tachibana Bay, Nagasaki, Japan, for different periods of time from November 2003 to January 2005. The effects of age, immersion month, dry weight, bacterial and diatom densities of these biofilms on the settlement and metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis were investigated in the laboratory. Furthermore, biofilms were subjected to various treatments to investigate the nature of the settlement and metamorphosis cue in the biofilm. Pediveliger larvae of the mussel settled and metamorphosed in response to biofilms. Settlement and metamorphosis to the post-larval stage significantly increased with the biofilm age. In addition, the biofilm activity varied depending on the immersion month (season), e.g., for biofilms with the same age, those immersed between June and August had higher activities than those immersed between November and March. The activity of the biofilm also positively correlated with the dry weight, bacterial and diatom densities. These three quantitative parameters of the biofilm were significantly affected by the film age but were not affected by the immersion month, suggesting that other parameters (e.g., community structures, extracellular products) also affected the inductive activity of the biofilm. The fixative agents (formalin and glutaraldehyde), heat, ethanol, ultraviolet irradiation and antibiotics treatments of the biofilm resulted in significant reduction or loss of its inductive activity. The survival of bacterial cells in the treated films where activities were either reduced or lost also decreased significantly. No settlement and metamorphosis were obtained when larvae were exposed to the conditioned water of the biofilm. Thus, larvae of M. galloprovincialis settled and metamorphosed in response to a cue produced by living bacteria in the biofilm. The cue may be a bacterial extracellular product which was susceptible to the above treatments.

Low-Intensity Photosensitization May Enhance RecA Production

Three bacterial strains-Escherichia coli, Acinetobacter calcoaceticus, and the A. calcoaceticus RecA- mutant-underwent photosensitization by a low-concentration (0.73 micromol/L) tetramethyl pyridyl porphine (a cationic hydrophylic photosensitizer) and a 4-J/cm2 dose of 407 to 420 nm blue light. The viability of the first two strains decreased by approximately 60%. and that of the RecA- strain decreased by 90%. Increasing the amount of photosensitizer to 14.6 micromol/L at the same dose of blue light resulted in a 95% to 98% decrease in viability of the three strains. Very little damage to the bacterial DNA was observed after this treatment. Increasing the concentration photosensitizer under the same illumination conditions also resulted in very little damage to the DNA. Western blotting demonstrated that the low photosensitization procedures enhance RecA production for mending the damaged chromosomal DNA. RecA production as a result of low-dose photosensitization was confirmed and demonstrated by immunofluorescent staining and gold immunolabeling. Although DNA is not the primary target for photosensitization, this process of RecA production may provide a certain degree of DNA mending and may also affect the survival of bacterial cells on low-intensity photosensitization.

Genome scale analysis of diffusible signal factor regulon in Xanthomonas campestris pv. campestris: identification of novel cell–cell communication‐dependent genes and functions

The bacterial pathogen Xanthomonas campestris pv. campestris (Xcc) recruits a diffusible signal factor (DSF), which has recently been structurally characterized as cis-11-methyl-2-dodecenoic acid, as a cell-cell communication signal to synchronize virulence gene expression and biofilm dispersal. In this study, we showed that despite the existance of phenotype variations in different Xcc isolates, the DSF-mediated functions were in general conserved. To investigate the genomic profiles of DSF regulation, we designed and conducted oligomicroarray analysis by comparison of the gene expression patterns of wild-type strain XC1 and its DSF-deficient mutant XC1dF, as well as those of XC1dF in the presence or absence of DSF signals. The analyses led to identification of 165 genes, whose expression was significantly influenced by DSF signals. These genes encode proteins and enzymes belonging to at least 12 functional groups. In addition to those previously known DSF-dependent activities such as production of extracellular enzymes and extracellular polysaccharides, microarray analyses also revealed new functions mediated by DSF, such as flagellum synthesis, resistance to toxins and oxidative stress, and aerobic respiration. Phenotype analyses confirmed that DSF signalling contributed to resistance to toxin acriflavin and hydrogen peroxide, and to the survival of bacterial cells at different temperatures. We conclude that DSF cell-cell signalling is not only essential for co-ordinating the expression of virulence genes but also plays a vital role in keeping up the general competence of the pathogen in ecosystems.

A Salt-Bridge Motif Involved in Ligand Binding and Large-Scale Domain Motions of the Maltose-Binding Protein

The uptake of nutrients is essential for the survival of bacterial cells. Many specialized systems have evolved, such as the maltose-dependent ABC transport system that transfers oligosaccharides through the cytoplasmic membrane. The maltose/maltodextrin-binding protein (MBP) serves as an initial high-affinity binding component in the periplasm that delivers the bound sugar into the cognate ABC transporter MalFGK2. We have investigated the domain motions induced by the binding of the ligand maltotriose into the binding cleft using molecular dynamics simulations. We find that MBP is predominantly in the open state without ligand and in the closed state with ligand bound. Oligosaccharide binding induces a closure motion (30.0° rotation), whereas ligand removal leads to domain opening (32.6° rotation) around a well-defined hinge affecting key areas relevant for chemotaxis and transport. Our simulations suggest that a “hook-and-eye” motif is involved in the binding. A salt bridge between Glu-111 and Lys-15 forms that effectively locks the protein-ligand complex in a semiclosed conformation inhibiting any further opening and promoting complete closure. This previously unrecognized feature seems to secure the ligand in the binding site and keeps MBP in the closed conformation and suggests a role in the initial steps of substrate transport.

Surface Characteristics of Spacecraft Components Affect the Aggregation of Microorganisms and May Lead to Different Survival Rates of Bacteria on Mars Landers

Layers of dormant endospores of Bacillus subtilis HA101 were applied to eight different spacecraft materials and exposed to martian conditions of low pressure (8.5 mbar), low temperature (-10 degrees C), and high CO(2) gas composition and irradiated with a Mars-normal ultraviolet (UV-visible- near-infrared spectrum. Bacterial layers were exposed to either 1 min or 1 h of Mars-normal UV irradiation, which simulated clear-sky conditions on equatorial Mars (0.1 tau). When exposed to 1 min of Mars UV irradiation, the numbers of viable endospores of B. subtilis were reduced three to four orders of magnitude for two brands of aluminum (Al), stainless steel, chemfilm-treated Al, clear-anodized Al, and black-anodized Al coupons. In contrast, bacterial survival was reduced only one to two orders of magnitude for endospores on the non-metal materials astroquartz and graphite composite when bacterial endospores were exposed to 1 min of Mars UV irradiation. When bacterial monolayers were exposed to 1 h of Mars UV irradiation, no viable bacteria were recovered from the six metal coupons listed above. In contrast, bacterial survival was reduced only two to three orders of magnitude for spore layers on astroquartz and graphite composite exposed to 1 h of Mars UV irradiation. Scanning electron microscopy images of the bacterial monolayers on all eight spacecraft materials revealed that endospores of B. subtilis formed large aggregates of multilayered spores on astroquartz and graphite composite, but not on the other six spacecraft materials. It is likely that the formation of multilayered aggregates of endospores on astroquartz and graphite composite is responsible for the enhanced survival of bacterial cells on these materials.

Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy

The mechanosensitive channel of large conductance (MscL) plays an important role in the survival of bacterial cells to hypo-osmotic shock. This channel has been extensively studied and its sequence, structure and electrophysiological characteristics are well known. Here we present a method to visualise MscL in living bacteria using confocal microscopy. By creating a gene fusion between mscl and the gene encoding the green fluorescent protein (GFP) we were able to express the fusion protein MscL-GFP in bacteria. We show that MscL-GFP is present in the cytoplasmic membrane and forms functional channels. These channels have the same characteristics as wild-type MscL, except that they require more pressure to open. This method could prove an interesting, non-invasive, tool to study the localisation and the regulation of expression of MscL in bacteria.

Effect of heparin binding on Helicobacter pylori resistance to serum.

The objective of this study was to evaluate the effect of heparin binding to Helicobacter pylori cells on their survival in the presence of fresh rabbit serum with or without active complement components. Three H. pylori strains were compared and the amounts of heparin added reflected the physiological concentrations that can be found in animal tissues. No growth of H. pylori was noted in the presence of serum. Serum with or without active complement produced a reduction in c.f.u. for strains SPM 326, CCUG 17874(T) and SS1. However, addition of heparin resulted in increased survival of bacterial cells in serum with or without active complement. It appears that heparin binding to H. pylori can prevent bacterial cell death due to the alternative complement system. Heparin binding could also protect from heated serum (complement-inactivated), indicating protection from other serum components besides complement. In vivo, the process of heparin binding could possibly result in facilitated colonization due to a higher survival rate.