F.P. de França

Influence of oil contamination levels on hydrocarbon biodegradation in sandy sediment

The influence of oil concentration on hydrocarbon biodegradation in a sandy sediment was studied in polyvinyl chloride reactors (0.45 x 0.28 x 0.31 m) containing 76.8 kg of beach sand in natura, where the upper layer was artificially contaminated with petroleum. The oil-degrading microorganisms used consisted of a mixed culture named ND, obtained from landfarming and associated with indigenous microorganisms. On the 28th day of the process, the degradation in reactors containing sandy sediment contaminated with light Arabian oil and presenting an initial oil content of 14, 21 or 28 g kg-1 reached the following levels (%): 33.7, 32.9 and 28.9 for oil and grease; up to 88.3, 35.3 and 13.0 for C14-C26 n-alkanes; and 100, 61.3 and 59.4 for pristane, respectively. Phytane removal (37.1%) was only detected in the reactor contaminated with the lowest oil concentration studied. These results, together with the expressive bacterial growth observed (from 10(6) to 10(11) cfu g-1) give strong support to the argument that biodegradation was the dominant component of the remediation process. Susceptibility to biodegradation was inversely proportional to increasing oil contamination. The degradation of branched alkane: pristane was not repressed by the presence of n-alkanes.

Continuous ethanol production using yeast immobilized on sugar-cane stalks

Sugar-cane stalks, 2.0 cm long, were used as a support for yeast immobilization envisaging ethanol production. The assays were conducted in 38.5 L fermenters containing a bed of stalks with 50% porosity. The operational stability of the immobilized yeast, the efficiency and stability of the process, as well as the best dilution rate were evaluated. Molasses from demerara sugar production was used in the medium formulation. It was diluted to obtain 111.75 ± 1.51 g/L without any further treatment. Sulfuric acid was used to adjust the pH value to around 4.2. Every two days Kamoran HJ (10 ppm) or with a mixture containing penicillin (10 ppm) and tetracycline (10 ppm), was added to the medium. Ethanol yield and efficiency were 29.64 g/L.h and 86.40%, respectively, and the total reducing sugars (TRS) conversion was 74.61% at a dilution rate of 0.83 h-1. The yeast-stalk system was shown to be stable for over a 60 day period at extremely variable dilution rates ranging from 0.05 h-1 to 3.00 h-1. The concentration of immobilized cell reached around 109 cells/gram of dry sugar-cane stalk when the fermenter was operating at the highest dilution rate (3.00 h-1).

Alleviation of water stress effects in cowpea by Bradyrhizobium spp. inoculation

Experiments were carried out to investigate the effects of different degrees of water stress on cowpea in the presence and absence of Bradyrhizobium spp. inoculation and to evaluate physiological responses to stress. The soil used was Yellow Latosol, pH 6.3 and the crop used was cowpea (Vigna unguiculata (L.) Walp.) cv. ‘IPA 204’. Stress was applied continuously by the control of matric potential (ψm) through a porous cup. The lowered soil ψm had a direct effect on the N2 fixation, but the strains Bradyrhizobium introduced by inoculation in the cowpea plants were superior to the indigenous strain demonstrating the importance of inoculation in the stressed plants. At the more negative ψm plants inoculated with the strains EI 6 formed associations of greater symbiotic efficiency which helped the cowpea plants to withstand drought stress better than the strain BR 2001 and the uninoculated control. The leghaemoglobin concentration was not inhibited in the drought-stressed plants at ψm -70 kPa when inoculated with the strain EI 6, which confered a differential degree of drought resistance in plants. The ψw declined in the stressed plants reaching values of -1.0 MPa which was sufficient to cause disturbance in nodulation and biomass production.

Optimizing Carbon/Nitrogen Ratio for Biosurfactant Production by a Bacillus subtilis Strain

A Bacillus subtilis strain isolated from contaminated soil from a refinery has been screened for biosurfactant production in crystal sugar (sucrose) with different nitrogen sources (NaNO3, (NH4)2SO4, urea, and residual brewery yeast). The highest reduction in surface tension was achieved with a 48-h fermentation of crystal sugar and ammonium nitrate. Optimization of carbon/nitrogen ratio (3, 9, and 15) and agitation rate (50, 150, and 250 rpm) for biosurfactant production was carried out using complete factorial design and response surface analysis. The condition of C/N 3 and 250 rpm allowed the maximum increase in surface activity of biosurfactant. A suitable model has been developed, having presented great accordance experimental data. Preliminary characterization of the bioproduct suggested it to be a lipopeptide with some isomers differing from those of a commercial surfactin.

Thermophilic and mesophilic bacteria in biofilms associated with corrosion in a heat exchanger

Biofilm development on AISI-1020 carbon steel coupons installed at the outlet of a heat exchanger was evaluated at the thirtieth and the sixtieth days of exposure. Water temperature varied between 41 and 60 °C. The most probable number technique (MPN) was applied to quantify mesophilic and thermophilic species of aerobic, anaerobic, and sulphate-reducing bacteria (SRB) in planktonic and sessile phases. The results showed predominance of thermophilic aerobic bacteria in both phases, corresponding to 9.5 ± 0.8 × 106 cells/ml in the planktonic phase. In biofilms, maximal aerobic cell concentration, 7.8 ± 0.6 × 108 cells/cm2, was registered at the sixtieth day. An increase in the number of thermophilic anaerobic and SRB with elapsed time was also observed. The results obtained after 60 days were 5.8 ± 0.4 × 107 and 8.9 ± 0.9 × 104 cells/cm2 for anaerobic and sulphate-reducing bacteria, respectively. Scanning electron microscopy showed a varied composition of species in the biofilms and corrosion on the carbon steel surfaces after biofilm removal.

Biofilm formation in water cooling systems

Biofilm formation on stainless steel samples immersed in cooling water has been evaluated by exposing metal samples to cooling seawater for 30 days. Anaerobic bacteria were then at 1.6 × 106/cm2, with sulphate-reducing species predominating. Aerobic bacteria and fungi were 2600 and 140/cm2, respectively. After 60 days, numbers of aerobic microorganisms remained constant whereas the count of anaerobic microorganisms had increased to 1.8×109/cm2. Scanning electron microscopy showed the presence of morphologically different microorganisms in deposits and as a mucilaginous net. No signs of corrosion were detected on the stainless steel surface.

Cadmium uptake by Spirulina maxima: toxicity and mechanism

Cadmium uptake by Spirulina maxima cells was more pronounced in living than in dead cells, with a maximum recovery of 47.63mg Cd/g cells for living cells and 37.00mg Cd/g cells for inactivated cells. When in the medium at 1.2mg/l, cadmium affected cell growth and diminished cell productivity. Cadmium was detected in the outer and inner faces of the external membrane, essentially in the lipid layer.Cadmium uptake by Spirulina maxima cells was more pronounced in living than in dead cells, with a maximum recovery of 47.63mg Cd/g cells for living cells and 37.00mg Cd/g cells for inactivated cells. When in the medium at 1.2mg/l, cadmium affected cell growth and diminished cell productivity. Cadmium was detected in the outer and inner faces of the external membrane, essentially in the lipid layer.

Biofilm formation on brass coupons exposed to a cooling system of an oil refinery

Brass coupons (70% Cu 30% Zn) were exposed to a cooling freshwater system of an oil refinery, in order to investigate susceptibility of the metal to biofilm formation. The coupons were fixed on bypasses at points which allowed the circulation of makeup, cooling and return water. The number of aerobic, anaerobic and sulfate-reducing bacteria was determined in both the planktonic and the sessile phases. Maximum bacterial concentrations were detected in the cooling water, corresponding to 2.1 ± 0.1 × 106 CFU ml−1 (planktonic phase) and 1.3 ± 0.2 × 105 CFU cm−2 (sessile phase) for aerobic bacteria and to 3.2 ± 0.3 × 105 cells ml−1 (planktonic phase) and 6.2 ± 0.7 × 105 cells cm−2 (sessile phase) for anaerobic bacteria. Sulfate-reducing bacteria (SRB) were observed only in the planktonic phase, being found in greater numbers in the return water. Scanning electron microscopy (SEM) analysis indicated that biofilm formation occurred at the three monitored sites and showed a diversity in cell morphology. Nonetheless, no evidence of corrosion was observed on the brass coupons during the experimental period.

Variation in sessile microflora during biofilm formation on AISI-304 stainless steel coupons

Coupons of stainless steel type AISI-304 were exposed to the industrial cooling system of a petrochemical plant fed by seawater from the Guanabara Bay, Rio de Janeiro, Brazil, in order to study thein situ formation of biofilms. Bacteria, microalgae and fungi were detected on the coupons as soon as 48 h after exposure. Their respective numbers were determined at times 48, 96 and 192 h and over the following 8 weeks. Aerobic, anaerobic and sulfate-reducing bacteria were quantified according to the technique of the most probable number, and fungi by the pour plate technique. The number of microorganisms present in the forming biofilm varied over the experimental period, reaching maximal levels of 14×1011 cells cm−2, 30×1013 cells cm−2, 38×1011 cells cm−2 and 63×105 cells cm−2, respectively, for aerobic bacteria, anaerobic bacteria, sulfate-reducing bacteria and fungi, and the dynamics of this variation depended on the group of microorganisms.Bacillus sp,Escherichia coli, Serratia sp andPseudomonas putrefaciens were identified among the aerobic bacteria isolated. Additionally, microalgae and bacteria of the genusGallionella were also detected. Nonetheless, no evidence of corrosion was found on the stainless steel type AISI-304 coupons over the experimental period.

Effect of different salinities of a dynamic water system on biofilm formation

AISI-1020 carbon steel coupons were fixed onto a water circulation loop in order to study the effect of varying NaCl concentrations on formation of biofilms by natural populations of microorganisms. Overall, we observed a reduction in the number of bacteria attached to the metal surfaces as NaCl levels increased. At 12.85 and 80 g/l NaCl, the respective bacterial counts were: 1.7×109 CFU/cm2 and 7.5×102 CFU/cm2 for aerobic species; 1.3×104 CFU/cm2 and 2.1×10 CFU/cm2 for anaerobic species; and 1.8×103 CFU/cm2 and 4.6×10 CFU/cm2 for sulfate-reducing species. However, the opposite trend was observed for the numbers of iron-reducing bacteria: 4.1×106 CFU/cm2 at 12.85 g/l NaCl and 7.5 108 CFU/cm2 at 80 g/l NaCl, respectively. Fungal counts remained constant throughout the experimental period. The salt concentration at which the maximum corrosion rate was observed was 35 g/l. In view of the marked loss of metal mass recorded at this salinity, AISI-1020 carbon steel proved to belong to the group of alloys less resistant to corrosion. Journal of Industrial Microbiology & Biotechnology (2000) 25, 45–48.