Implication of Mn-cofactored superoxide dismutase in the tolerance of swarmer Pseudomonas aeruginosa to polymixin, ciprofloxacin and meropenem antibiotics.

The protective role of superoxide dismutase (Sod) against oxidative stress, resulting from the common antibiotic pathway of action, has been studied in the wild type and mutant strains of swarmer Pseudomonas aeruginosa, lacking Cytosolic Mn-Sod (sodM), Fe-Sod (sodB) or both Sods (sodMB).Our results showed that inactivation of sodB genes leads to significant motility defects and tolerance to meropenem. This resistance is correlated with a greater membrane unsaturation as well as an effective intervention of Mn-Sod isoform, in antibiotic tolerance.Moreover, loss of Mn-Sod in sodM mutant, leads to polymixin intolerance and is correlated with membrane unsaturation. Effectivelty, sodM mutant showed an enhanced swarming motility and a conserved rhamnolipid production. Whereas, in the double mutant sodMB, ciprofloxacin tolerance would be linked to an increase in the percentage of saturated fatty acids in the membrane, even in the absence of superoxide dismutase activity.The overall results showed that Mn-Sod has a protective role in the tolerance to antibiotics, in swarmer P.aeruginosa strain. It has been further shown that Sod intervention in antibiotic tolerance is through change in membrane fatty acid composition.

Pseudomonas aeruginosa Swarmer Cells Adaptation Toward UVc Radiations.

Swarming is the most rapid surface motility allowing Pseudomonas aeruginosa bacteria to rapidly colonize new surfaces. However, swarming behavior is affected by environmental factors like ultraviolet irradiation (UVc). UVc radiation is the most disinfection technology usually applied for wastewater and proven to be effective to inactivate microorganisms. However, efficiency against motile bacteria is not yet studied. This study aims to explain the mechanisms of resistance of swarmer P. aeruginosa cells toward UVc exposure. P. aeruginosa liquid cultures were allowed to swarm across a semisolid surface for 18 h and directly exposed to UVc radiations. Emergent swarmer colonies, revealed after re-incubation, were selected to study biofilm formation, fatty acid (FA) composition, and ultrastructure. Our results showed that membrane adaptation to UVc radiations was seen in Pseudomonas cells by an increase of cyclic fatty acid (CFA) content, confirming the role of cyclopropane in radio-resistance of swarmer cells. Furthermore, electron microscopic study confirmed that over production of S-layer is believed to be a protective form adopted by P. aeruginosa swarmer cells to resist after 5 min of UVc exposure. Moreover, membrane disintegration is the lethal effect observed after 15 min of UVc exposure. In the other hand, study of biofilm production showed an enhancement of biofilm formation, of swarmer cells mainly after 15 min of UVc exposure. There results confirmed that swarming process is highly correlated with particular FA composition of P. aeruginosa membrane and that radio-resistance of swarmer cells is highly supported by CFA biosynthesis and S-layer overproduction.

Antioxidant Defense Mechanisms in Pseudomonas aeruginosa: Role of Iron-Cofactored Superoxide Dismutase in Response to UV-C Radiations.

The role of SOD gene in response to UV-C radiations was studied in Pseudomonas aeruginosa. Firstly, our results showed that the inactivation of sodM and/or sodB genes decreases the resistance of P. aeruginosa after exposure to UV-C rays. Furthermore, our results showed that SOD activity is dose dependant in all strains. However, significant increase in SOD activity was only shown at UV-C exposure time of 5 min in sodB mutant. At an elevated dose equivalent to 30 min of exposure, significant increase in SOD activity was observed in sodM. Catalase activities showed significant decrease in WT and in sodB mutant after an exposure time of 30 min. CAT enzyme was present at higher levels than SOD, reflecting that alternate enzymes such as POX, is poorly associated with CAT activity, and an increase in POX activity is related to increase in stress tolerance. The overall results showed that sodB gene has an important protective role against UV-C radiations in P. aeruginosa, compared to SodM isoform.

UV-C pre-adaptation of Salmonella: effect on cell morphology and membrane fatty acids composition.

The present study was carried out to evaluate the effects of ultraviolet radiations (UV-C) on the fatty acids composition of three serovars of Salmonella: S. typhimurium, S. hadar and S. zanzibar. Results obtained show that UV-C treatment increases significantly (P ≤ 0.05) the percentage of cyclic fatty acids. The atomic force microscopy was used to study the morphology and cell surface of irradiated strains. Results show that UV-C rays induce morphological changes and alter the bacterial cell surface (presence of grooves and irregularities).

Allelic polymorphisms of human platelets-specific alloantigens in South Tunisian population.

OBJECTIVES: Human platelet-specific alloantigens (HPA) are polymorphic epitopes which vary among ethnic groups. BACKGROUND: In Tunisia, HPA frequencies were determined in North and centre; however, the pattern of HPA in South Tunisian population is not been studied yet. The aim of this work was to determine allelic frequencies of HPA-1, -3, and -5 systems in south Tunisian population, in order to estimate the risk of anti-platelet allo-immunization and to create a register of HPA-typed blood donors. METHODS: Our study concerned 212 unrelated healthy, regular blood donors from southern Tunisia. Allelic polymorphisms of each system were determined using a polymerase chain reaction with sequence-specific primers. RESULTS: Genotype frequencies a/a, a/b, and b/b were, respectively, 0.670, 0.288, and 0.042 for HPA-1 system, 0.430, 0.462, and 0.108 for HPA-3 system, and 0.750, 0.241, and 0.009 for HPA-5 system. The allele frequencies were 0.814 and 0.186 for HPA-1a and -1b alleles; 0.660 and 0.340 for HPA-3a and -3b alleles and 0.870, and 0.130 for HPA-5a and -5b alleles. DISCUSSION: The reported frequencies are more similar to those of Caucasians than those of north Tunisian population.

Changes in membrane fatty acid composition of Pseudomonas aeruginosa in response to UV-C radiations.

The changes in lipid composition enable the micro-organisms to maintain membrane functions in the face of environmental fluctuations. The relationship between membrane fatty acid composition and UV-C stress was determined for mid-exponential phase and stationary phase Pseudomonas aeruginosa. The total lipids were obtained by dichloromethane/methanol (3:1) and were quantified by GC. The TLC analysis of phospholipids showed the presence of three major fractions phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. Significant modifications, as manifested by an increase of UFA, were obtained. Interestingly, this microorganism showed a remarkable capacity for recovery from the stressful effects of UV-C.

Prevalence of factor V Leiden in south Tunisian blood donors.

Venous thrombosis (VT) is a common disease with multifactorial pathogenesis. Factor V Leiden mutation (G1691A) (FVL) is the most common risk factor in venous thrombosis. The prevalence of FVL varies according to geography and ethnicity. Hence, in several countries there is a difference in the frequency of this mutation between the southern, central and north. In Tunisia, no data is available about prevalence of FVL mutation by geographical origin. For this reason, we sought the prevalence of FVL mutation in blood donor of south Tunisia population. FVL has been detected by APCR-test and confirmed by PCR-RFLP and sequencing. Two hundred fifty blood donors, different in age and sex were included in this study to determine the prevalence of FVL in blood donors. FVL mutation was found in 13.6% of the studied population. Thirty-one were heterozygous and three persons were homozygous with a rate of 12.4 and 1.2%, respectively. In conclusion, FVL mutation is very common in south Tunisian population.