Surface Properties of Bacteria from Different Wastewater Treatment Plants

The surface properties of the individual members of degradative biocommunities isolated from different laboratory and natural populations were characterized. The bacterial strains isolated from a given origin and degrading a given substrate varied with respect to their hydrophobic and electrostatic properties (e.g. contact angle, adsorption to hexadecane, isoelectric point, adsorption of anionic orcationic dyes). However, despite their specific surface characteristics, in most cases the net charge properties of different bacterial strains (characterized by the zeta potential profiles of the bacteria in relation to the pH) were found to be related to the substrate the bacteria were able to degrade as well as to the consortium the bacteria were isolated from. For one group of specialized bacteria, only oneor at most two characteristic zeta potential profiles were measured. Compared to the differences between different strains, the zeta potential profiles of individual strains were only slightly affected by either growth state or changes in the actual nutrient composition. Even if isolated strains were cultivated in standard nutrient broth for several months, only slight differences in the zeta potential profiles were measured. Only the isoelectric focusing experiments indicated thatcultivation in a complex medium favoured a progressively decreased uniformity of surface charge properties. Thus, measurement of zeta potential profiles under standardized conditions may be a useful means to compare the surface structures of bacteria from different origins.     

The influence of saliva on the hydrophobic surface properties of bacteria isolated from oral sites of macaque monkeys

Abstract The surface hydrophobicity of 64 bacterial strains isolated from discrete, intra-oral sites of monkeys ( Macaca fascicularis ) was determined by measuring their affinity for hexadecane. Bacteria were also exposed to monkey saliva which either increased or reduced the surface hydrophobicity of the cells. After exposure to saliva those bacteria isolated solely from the mucosal surfaces were significantly more hydrophobic than bacteria ( Streptococcus mutans and Actinomyces spp.) whose major habitat was the dentition. Streptococcus sanguis strains isolated from all intra-oral sites and among the early plaque formers were as hydrophobic as the organisms isolated only from the mucosal surfaces.     

The effect of culture conditions on hydrophobic properties of Pseudomonas aeruginosa

The cell surface hydrophobicity (CSH) plays an important role in a adhesion of bacteria on solid surfaces. CSH of 62 Pseudomonas aeruginosa strains isolated from humans and different animals was assessed using the ammonium sulfate salt aggregation test. Bacteria were grown for 24 h and 48 h at a room temperature (22 degrees C) and 37 degrees C on enrichment broth and agar (Biomed) and tryptic soy agar (Difco). The hydrophobic properties of the Pseudomonas aeruginosa strains were depended on the temperature, time of the culture of bacteria and the kind of media. CSH properties were most frequently expressed when the analyzed strains were cultured in enrichment broth. In a such conditions Pseudomonas aeruginosa strains were more hydrophobic when grown at 22 degrees C (94% after 24 h and 87% after 48%) than those at 37 degrees C (72% after 24 h and 71% after 48 h). Among strains cultured in tryptic soy agar at 37 degrees C, 48% after 24 h and 75% after 48 h were autoaggregating, representing very strong hydrophobic properties.     

Evaluation of hydrophobic properties of Yersinia enterocolitica strains isolated from humans and pigs]

The aim of the study was to evaluate the hydrophobic properties of Yersinia enterocolitica and to determine the influence of the culture conditions, such as: type of medium, temperature, and duration of the culture on the manifestation of these properties. The subject of the study were 117 of Y. enterocolitica strains isolated from humans and pigs. The ammonium sulphate salt aggregation test according to Lindahl modified method was used to evaluate the hydrophobic properties of Y. enterocolitica strains. Strains of Y. enterocolitica were cultured for 24 h at 25 degrees C on TSA (Difco) medium. During investigation of the influence of the culture conditions the chosen strains were incubated for 24 h and 48 h at 25 degrees C and 37 degrees C on TSA (Difco), LB (Difco), enrichment agar (Biomed), and enrichment agar with 5% sheep blood (Graso). A total of 44.5%, 17.9%, 9.4%, and 28.2% strains of Y. enterocolitica showed very strong hydrophobic properties, strong hydrophobic properties, some hydrophobic properties, and were non-hydrophobic, respectively when strains of Y. enterocolitica were cultured for 24 h at 25 degrees C on TSA medium. A total of 75.5% strains isolated from humans showed very strong hydrophobic properties and 13.5% strains were non-hydrophobic. Among strains isolated from pigs 30% showed very strong hydrophobic properties but 35% were non-hydrophobic. The hydrophobic properties of Y. enterocolitica depended on the temperature, duration of the culture and the type of media. The highest number of strains with very strong hydrophobic properties (89.6%) was obtained after 48 h of the incubation at 37 degrees C on the enrichment agar with 5% sheep blood. The highest number of non-hydrophobic strains of Y. enterocolitica (28.5%) was obtained after 24 h at 25 degrees C on TSA medium.     

Effect of avian intestinal microflora possessing adhering and hydrophobic properties on competitive exclusion of Salmonella typhimurium from chicks

The role of adherence and hydrophobic properties of native gut microflora in competitive exclusion of salmonellas from chicks is reported. Pure bacterial strains were isolated from washed caeca of 3-d-old chicks which had been treated on the day of hatch with a microflora from salmonella-free adult birds. These strains, when added to known mixtures of pure cultures, improved the efficacy of the mixtures in protecting chicks against a challenge of 10⁵ cfu of Salmonella typhimurium. Strains from washed caeca and from other sources were also screened for hydrophobic properties. Undefined microflora and strains freshly isolated from washed caeca and which were hydrophobic improved the efficacy of protective mixtures, while none of the combination of hydrophobic strains obtained from stored strains had any beneficial effect.     

Effect of avian intestinal microflora possessing adhering and hydrophobic properties on competitive exclusion of Salmonella typhimurium from chicks.

The role of adherence and hydrophobic properties of native gut microflora in competitive exclusion of salmonellas from chicks is reported. Pure bacterial strains were isolated from washed caeca of 3-d-old chicks which had been treated on the day of hatch with a microflora from salmonella-free adult birds. These strains, when added to known mixtures of pure cultures, improved the efficacy of the mixtures in protecting chicks against a challenge of 10(5) cfu of Salmonella typhimurium. Strains from washed caeca and from other sources were also screened for hydrophobic properties. Undefined microflora and strains freshly isolated from washed caeca and which were hydrophobic improved the efficacy of protective mixtures, while none of the combination of hydrophobic strains obtained from stored strains had any beneficial effect.     

Surface properties of Staphylococcus aureus affecting chemiluminescence response of human phagocytes

To assess the surface properties of Staphylococcus aureus affecting the response of human phagocytes, the effects of the organisms with different surface properties on the chemiluminescence (CL) response of human phagocytes were examined. The magnitude of the phagocytic CL response to hydrophobic strains was significantly greater than that to hydrophilic strains, while no significant difference in the CL response was seen between protein A-deficient strains and their parent strains. The CL response to the hydrophilic organisms prepared from a hydrophobic strain by trypsin treatment decreased significantly. These results suggest that the phagocytic CL response to staphylococci depends on the hydrophobicity of the surface, but not on the presence of protein A. Two protein A-deficient strains which were isolated from protein A-positive strains showed identical hydrophobicity with their parent strains. All of the hydrophilic strains isolated from hydrophobic strains possessed protein A identical to that of their parent strains. Moreover, a hydrophilic strain could be isolated from a protein A-deficient, hydrophobic strain. These results strongly suggest that protein A is not solely responsible for the surface hydrophobicity of S. aureus.     

Surface properties of lactobacilli isolated from the small intestine of pigs

One hundred wild-type strains of the genus Lactobacillus were isolated from the small intestine of newly-slaughtered pigs up to 6 months of age. Cell surface hydrophobicity and capsule formation were studied on a number of strains. Strains showing high surface hydrophobicity as measured by the salt-aggregation test and hydrophobic interaction chromatography on Octyl Sepharose were commonly found to adhere in high numbers to isolated pig intestinal epithelial cells. Heat and protease treatment of bacteria of high surface hydrophobicity, including autoaggregating strains in phosphate-buffered saline, showed a drastic decline in this surface property. Three hydrophilic strains (LBp 1044, 1068 and 1073) also showed binding to intestinal cells but at a lower level (approx. 5 bacteria/cell) as compared with the best binding hydrophobic strain (LBp 1063, approx. 11 bacteria/cell). These findings suggest that different or multiple adhesion mechanisms may be involved in the colonization of the small intestinal mucosa of pigs. Cultures of selected strains grown in liquid media rich in carbohydrates did not affect their hydrophobic cell surface character. Therefore it seems less likely that carbohydrate capsule polymers are the major determinants of intestinal colonization of lactobacilli in pigs.     

Surface properties of lactobacilli isolated from the small intestine of pigs

One hundred wild-type strains of the genus Lactobacillus were isolated from the small intestine of newly-slaughtered pigs up to 6 months of age. Cell surface hydrophobicity and capsule formation were studied on a number of strains. Strains showing high surface hydrophobicity as measured by the salt-aggregation test and hydrophobic interaction chromatography on Octyl Sepharose were commonly found to adhere in high numbers to isolated pig intestinal epithelial cells. Heat and protease treatment of bacteria of high surface hydrophobicity, including autoaggregating strains in phosphate-buffered saline, showed a drastic decline in this surface property. Three hydrophilic strains (LBp 1044, 1068 and 1073) also showed binding to intestinal cells but at a lower level (approx. 5 bacteria/cell) as compared with the best binding hydrophobic strain (LBp 1063, approx. 11 bacteria/cell). These findings suggest that different or multiple adhesion mechanisms may be involved in the colonization of the small intestinal mucosa of pigs. Cultures of selected strains grown in liquid media rich in carbohydrates did not affect their hydrophobic cell surface character. Therefore it seems less likely that carbohydrate capsule polymers are the major determinants of intestinal colonization of lactobacilli in pigs.     

The hemagglutinating properties of Pseudomonas aeruginosa strains isolated from patients with nonspecific lung diseases]

To detect d-mannose-sensitive (MS) pili in 31 P. aeruginosa strains isolated from the respiratory tract of patients with inflammatory and purulent destructive pulmonary diseases, the hemagglutination (HA) test was used. The isolated Pseudomonas under study differed in the degree of manifestation of their MS adhesins. Among them microorganisms with pronounced HA activity (high HA titer) occurred, as well as those whose HA activity was less pronounced (low HA titer). P. aeruginosa strains with pronounced HA activity were more frequently isolated from patients with purulent destructive processes in the lungs. A correlation between the state of the patient at the moment of bacteriological examination and the degree of manifestation of MS pili in the P. aeruginosa strain isolated from this patients was established. The value of HA titer in the presence of d-mannose is indicative of the presence of MS adhesins in a P. aeruginosa strain.     

Surface Properties of Bifidobacterial Strains of Human Origin

The adherence of Bifidobacterium strains isolated from infant feces and commercial fermented dairy products to enterocyte-like cells was correlated with the autoagglutination and hemagglutination properties of these organisms. These results allowed us to define two groups: (i) cell-adherent bacteria showing hemagglutination and autoagglutination and (ii) non-cell-adherent, nonhemagglutinating, nonautoagglutinating bacteria. Glass adherence was shown to be nonspecific and was discarded as a criterion for selection of adherent cells. Hydrophobicity appeared to be necessary for adhesion to enterocyte-like cells and autoagglutination. Adhesive strains were highly hydrophobic, and the degree of adherence was slightly dependent on the surface potential. Cells autoagglutinated more when the electrostatic negative charges on the cell surface were shielded by a decrease in the pH from 7 to 2. However, in some strains negative charges at the cell surface were adjuvant to adhesion, thus suggesting that specific chemical interactions occurred. The present results provide a method for preliminary selection of bacteria potentially adherent to epithelial cells by means of autoagglutination.     

Identification of Bacterial Strains Isolated from the Mediterranean Sea Exhibiting Different Abilities of Biofilm Formation

The Mediterranean Sea has rarely been investigated for the characterization of marine bacteria as compared to other marine environments such as the Atlantic or Pacific Ocean. Bacteria recovered from inert surfaces are poorly studied in these environments, when it has been shown that the community structure of attached bacteria can be dissimilar from that of planktonic bacteria present in the water column. The objectives of this study were to identify and characterize marine bacteria isolated from biofilms developed on inert surfaces immersed in the Mediterranean Sea and to evaluate their capacity to form a biofilm in vitro. Here, 13 marine bacterial strains have been isolated from different supports immersed in seawater in the Bay of Toulon (France). Phylogenetic analysis and different biological and physico-chemical properties have been investigated. Among the 13 strains recovered, 8 different genera and 12 different species were identified including 2 isolates of a novel bacterial species that we named Persicivirga mediterranea and whose genus had never been isolated from the Mediterranean Sea. Shewanella sp. and Pseudoalteromonas sp. were the most preponderant genera recovered in our conditions. The phenotypical characterization revealed that one isolate belonging to the Polaribacter genus differed from all the other ones by its hydrophobic properties and poor ability to form biofilms in vitro. Identifying and characterizing species isolated from seawater including from Mediterranean ecosystems could be helpful for example, to understand some aspects of bacterial biodiversity and to further study the mechanisms of biofilm (and biofouling) development in conditions approaching those of the marine environment.     

Pore-forming properties and antibacterial activity of proteins extracted from epidermal mucus of fish.

Among several biological functions, the epidermal mucus of fish may play an important role in host defense, particularly in the prevention of colonization by parasites, bacteria and fungi. In previous work, two hydrophobic proteins of 27 and 31 kDa were isolated from carp mucus. This study identified a strong antibacterial activity (0.16-0.18 microM) well correlated with pore-forming properties. Here this work was extended to other fish species, four fresh water fish and one sea water fish. After a first step of purification, water-soluble and hydrophobic material were separated, and both fractions were analyzed by SDS-PAGE and capillary electrophoresis. Only the hydrophobic component induced pore-forming activity, when reconstituted in planar lipid bilayers. This pore-forming activity was well correlated to a strong antibacterial activity against several bacteria strains. These results suggest that fish secrete antibacterial proteins able to permeabilize the membrane of the target cell and thus act as a defense barrier.     

Study of beta-lactamase activity of staphylococci from different sources

The beta-lactamase activity of staphylococci isolated from the nasopharynx and skin of children with destructive affections of the lungs and from blood of patients with cardiovascular diseases subjected to surgical operations was determined with acidometric and microbiological procedures. Interrelation between synthesis of beta-lactamase by the staphylococcal strains and their resistance to beta-lactam antibiotics was demonstrated. No correlation of the antibiotic resistance and the taxonomic position of the staphylococcal strains was observed.     

Bacterial biofilm in seawater: cell surface properties of early-attached marine bacteria

The development of antifouling strategies in seawater requires knowledge of the physico-chemical properties of the cell surfaces of early adherent bacteria. The hydrophilic, electrostatic and the Lewis acid-base cell surface properties of eleven marine bacteria were characterized. Although these bacteria adhered to a hydrophilic support immersed for 3 and 6 h, they presented various physico-chemical properties. Eleven strains possessed a hydrophilic surface and five a hydrophobic surface. Although the majority of the bacteria presented an electron-donating character, some could not generate Lewis acid-base interactions with the support. On the other hand, all strains possessed an isoelectric point ranging from 2.2 to 3.4 and were negatively charged at the pH of seawater. Hydrophilicity was a preponderant property among these bacteria, but other properties should not be ignored. The development of new antifouling paints must take account all the possible interaction levels used by the bacteria to adhere to an immersed surface.     

Autoaggregation,hydrophobic, and hydrophylic properties of Moraxella catarrhalis strains

The hydrophobicity of the bacterial cell surface was evaluated via the salt aggregation test (SAT) in 58 strains (19 from the lower and 39 from the upper respiratory tracts) of Moraxella catarrhalis in hospitalized patients aged 25 to 65. Based on the SAT results, the strains were divided into three groups: autoaggregating (highly hydrophobic), hydrophobic, and hydrophilic. At a temperature of 37 degrees C, the autoaggregating, hydrophobic or hydrophilic properties did not depend on the choice of a medium, whereas at 22 degrees C the investigated properties did (p<0.0001). Taking into account the origin of the strains (lower vs. upper respiratory tract), it was found that: in the strains cultivated in liquid medium, both highly hydrophobic, hydrophobic and hydrophilic surfaces were present with a comparative frequency, independent of the strain isolation site and cultivation conditions; strains with highly hydrophobic and hydrophobic surfaces, but only those cultivated on solid media at 22 degrees C, were much more often isolated from sputum rather than from nose and throat swabs, whereas a statistically significant incidence of hydrophilic strains was found in samples from the upper rather than lower respiratory tract.     

Advances in the design of probiotic products for the prevention of major diseases in dairy cattle

Probiotics are frequently used for multiple objectives and the majority of applications are performed both in human beings and in different animals. The specificity of species showed by the members of the indigenous microbiotia led us to isolate lactic acid bacteria (LAB) from their natural bovine ecosystems and to select those with probiotic potentiality. The beneficial probiotic LAB were isolated from the vaginal tract and mammary gland of adult heifers, faeces and oral cavity of newborn calves. Bacterial strains and their numbers varied in each tract under study. Their surface properties, evaluated using the MATH test, revealed that the higher number of low hydrophobic strains were isolated from vaginal tract, mammary gland or faeces. The capability of autoaggregation also differed with higher degree in those isolated from vagina, whereas those that originated in the mammary gland or faeces showed low autoaggregation characteristics. The production of antagonistic substances showed that the production of hydrogen peroxide was higher in the strains isolated from the vaginal tract, while those able to produce organic acid were isolated from all the environments. A few strains were able to produce bacteriocins and were isolated only from the mammary gland and faeces, but not from the vaginal tract. Further studies are being performed to complete the functional characterization of the strains in order to evaluate their survival in the environmental conditions of each tract. Finally, the selected strains could be combined for the design of probiotic products to be used both in adult cows and in neonates, based on the host-specificity, due to their autochthonous origin and their potential use in the tract where they were isolated.     

Bacterial Biofilm in Seawater: Cell Surface Properties of Early-attached Marine Bacteria

The development of antifouling strategies in seawater requires knowledge of the physico-chemical properties of the cell surfaces of early adherent bacteria. The hydrophilic, electrostatic and the Lewis acid-base cell surface properties of eleven marine bacteria were characterized. Although these bacteria adhered to a hydrophilic support immersed for 3 and 6?h, they presented various physico-chemical properties. Eleven strains possessed a hydrophilic surface and five a hydrophobic surface. Although the majority of the bacteria presented an electron-donating character, some could not generate Lewis acid-base interactions with the support. On the other hand, all strains possessed an isoelectric point ranging from 2.2 to 3.4 and were negatively charged at the pH of seawater. Hydrophilicity was a preponderant property among these bacteria, but other properties should not be ignored. The development of new antifouling paints must take account all the possible interaction levels used by the bacteria to adhere to an immersed surface.     

Capsular polysaccharides secreted by building façade colonisers: characterisation and adsorption to surfaces

Exopolymers secreted by algal and cyanobacterial strains isolated from building fa?ades were imaged by microscopy techniques. They were extracted and characterised to investigate their possible contribution to interactions with solid surfaces. The polymers were polysaccharides, with anionic and hydrophobic properties varying between the various strains. Capsular polysaccharides extracted from a strain of Klebsormidium flaccidum adsorbed in higher amounts on hydrophobic than on hydrophilic surfaces. These results tend to confirm the hypothesis that exopolymers are important in the colonisation process of microorganisms to surfaces.     

Persistense of mono- and associated cultures of conditionally pathogenic enterobacteria

The comparative study of adhesive, hemolytic, DNA-ase, lecithinase, antilysozymic, anticomplementary activities of mono- and associated cultures of 57 Enterobacter spp., 61 Citrobacter spp. and 55 Serratia spp. strains, isolated from patients with pyoinflammatory, intestinal and urological diseases is carried out. Different variations of cocultivated bacteria including Enterobacter and Citrobacter, Enterobacter and Serratia, Citrobacter and Serratia are used. It was shown, that cocultivated Enterobacter, Citrobacter and Serratia bacteria increased the persistent properties of mixt cultures.