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The number of colony forming units (CFU) of different groups of bacteria and fungi in samples stored at temperatures of 5 to -12oC for 0-32 weeks was evaluated. The number of CFU obtained after the different periods of storage of red latossol soil was compared with the number of colonies obtained immediately after removal of soil samples (time zero). The number of total bacteria and actinomycetes in the samples remained practically unchanged throughout the storage period. The number of Gram-negative bacteria decreased by as much as 69% compared to control, while the number of Bacillus spp and of fungi increased 1.9 to 4.9 times starting from the 12th week in samples stored at 5oC. Except for the variations observed in fungal counts, the remaining groups of bacteria practically showed the same variation in number of colonies in soil samples stored at 5oC and -12oC.

In an attempt to screen out new potent antibiotic producers from soil, Streptomyces hygroscopicus D1.5 was isolated and was found antagonistic to both bacteria and fungi. It could utilise arginine as nitrogen source and glycerol as carbon source at 0.75 g/l and 11.5 g/l level, respectively for maximum antibiotic yield.

The electrophoretic profiles of penicillin binding proteins (PBPs) and outer membrane proteins (OMPs) of Yersinia pestis EV 76 were determined following in vivo growth in diffusion chambers implanted in the peritoneal cavity of mice. In contrast to Y. pestis grown under in vitro conditions which activate the low calcium response (LCR) regulon there was no significant qualitative or quantitative change of the PBP profile of Y. pestis cells during growth in diffusion chambers for up to 72 h following implantation in mice. Three OMPs, with molecular weight of 100, 60 and 58 kDa, were expressed in Y. pestis cells grown for 24 h, but not at 48 h or at 72 h, in diffusion chambers. These results indicate that growth of Y. pestis in intraperitoneal diffusion chambers activates genes which might be relevant to the growth in the mammal host.

Among one thousand eight hundred and thirty-four Gram-negative bacilli, isolated at Universidade Federal do Ceará hospital complex - Brazil, from January 1995 to February 1996, 456 (24.8%) were Nonfermentative Gram-Negative Bacilli (NFGNB). This study reports their identification to the species level and their frequency as well. Thirteen genera and thirty species were identified and Pseudomonas aeruginosa was the most frequent species (69.95%), followed by Acinetobacter baumannii (5.48%) and by Acinetobacter lwoffii (3.95%). Among the identified P.aeruginosa strains, 94.1% produced pigment but 7.9% of them produced pigment only after being cultivated several times. The frequency of the most species was similar to that reported in the literature.

Ribonuclease production by Aspergillus flavipes, A. sulphureus and A. fischeri in semi-synthetic medium, after 24-144 hours at 30ºC under shaking, was studied. After cultivation, the medium was separated from micelia by filtration and the resultant solution was used as enzymatic extract. The highest amount of biomass and RNase was obtained after 96 hours of cultivation. The enzymes produced by three species presented similar characteristics, with optimum temperature at 55ºC and two peaks of activity at pH 4.5 and 7.0. A. flavipes RNases were more sensitive to temperature: 50% of the initial activity was lost after 1 hour at 70ºC. After this heat treatment, RNase of A. sulphureus lost 30% of this activity and that of A. fischeri only 16%. The nucleotides released by enzimatic hydrolysis of RNA were separated by ion exchange chromatography in a AG-1X8-formiate column and identified by paper chromatography. This procedure indicated that the raw enzymatic extract of Aspergillus flavipes is able to hydrolyze RNA, releasing 3'-nucleotides monophosphate at pH 4.5 and 3' and 5'-nucleotides monophosphate at pH 7.0 and 8.5. This result suggests that this strain produces two different types of RNase, one acidic and other alcaline, with different specificities.

Viability of 6 mushroom strains of the Pleurotus genus (2 from P. djamor var. djamor, 1 from P. ostreatus var. ostreatus, 2 from P. ostreatus var. columbinus and 1 from P. pulmonarius) after liquid nitrogen cryopreservation (-196º) was evaluated. The contact time for the mycelia of these strains with the cryoprotectant (glycerol) was studied 1, 2 and 3 hours before freezing. We also tested the effect of different times (5, 10 and 15 minutes) and temperatures (30, 45 and 60ºC) of the thawing system for mycelial recovery. The results showed a marked tendency toward faster mycelial recovery when samples were thawed at 30ºC, while at 60ºC no recovery was observed. A change in thawing and contact times with the cryoprotectant did not affect the results significantly, as the thawing temperature and strain employed affected.

Curvularia pallescens Boedijn (Hyphomycetes) is redescribed with the aid of a scanning electron microscope, and the optimal cultural conditions for growing this fungus are discussed. Cytological analysis and nuclear condition, observed through the HCl-Giemsa technique, showed vegetative and reproductive structures (hypha and conidia) formed by uni, bi, tri, and multinucleated segments. Cultures of C. pallescens in Complete medium and in Potato dextrose agar varied on growth, on aspects of the border of the colonies and also on medium pigmentation. The Complete medium and the temperature between 25-28°C were the most indicated for growth of C. pallescens.

Pediococcus acidilactici (IL01) has grown in MRS (Man, Rogosa and Sharpe) broth modified by substitution of glucose by 2.0% (MRS-2), 3.0% (MRS-3), 4.0% (MRS-4) and 5.0% (MRS-5) sugar cane blackstrap molasses. The highest acid production was obtained in MRS-5 broth maintained at a constant pH of 5.0. The highest biomass production was obtained when P. acidilactici was grown in MRS-5 broth at initial pH 6.5, while productivity was higher in MRS-2 broth (28.16%). When the MRS-2 broth was utilized at initial pH 6.5 for a 20-hour fermentation period, the highest growth rate (dx/dt) was found in a period of 8 to 16 hours (0.290 g cells/L.h), while the specific growth rate (µ) was 0.175 (h-1) for that period, differently from the 0.441 (h-1) obtained for the period comprising the 4th to the 12th hour. The growth in MRS broth was 5.08% (2.95 g/l) higher than in MRS-2 broth (2.80 g/l). The data obtained have shown that P. acidilactici has had a significant growth in molasses as the main carbon source, and that it is possible to substitute MRS glucose by this carbon source with the purpose of obtaining a more economical growth medium for the potential large scale productions.

Solid state fermentations were carried out to test the efficacy of Ceratocystis fimbriata to grow on different agro-industrial substrates and aroma production. Seven media were prepared using cassava bagasse, apple pomace, amaranth and soya bean. All the media supported fungal growth. While amaranth medium produced pineapple aroma, media containing cassava bagasse, apple pomace and soya bean produced a strong fruity aroma. The aroma production was growth dependent and the maximum aroma intensity was detected a few hours before or after the maximum respirometric activity. Sixteen compounds were separated by gas cromatography of the components present in the headspace and fifteen of them were identified as acid (1), alcohols (6), aldehyde (1), ketones (2) and esters (5).

Prostaglandin A1 (PGA1) inhibits Mayaro virus replication in Aedes albopictus cells at nontoxic doses to uninfected cells. At 10 µg/ml, PGA1 decreases virus production by 90%. The presence of PGA1 during virus adsorption, with no treatment after infection, reduces virus yield by 41%. Antiviral activity is observed even when treatment starts at one or two hours post-infection. However, in cells pre-treated with PGA1 during 24 hours, virus replication is not impaired. Thus, events ocurring during initial stages of infection and after virus adsorption and penetration must be the target of PGA1 action. SDS-PAGE analysis of 35S-methionine labelled proteins shows that PGA1 inhibits the synthesis of viral proteins and induces the synthesis of polypeptides with molecular weight of 70 kDa, 57 kDa and 23 kDa. In cells pre-treated with actinomycin D the induction of those proteins is suppressed. In addition, actinomycin D treatment prevents PGA1antiviral activity, indicating that PGA1-induced stress proteins are probably involved in this mechanism.

Aeromonas has been described as an emergent foodborne pathogen of increasing importance. In this study, we report that 48% of 50 Pintado fish samples collected at the retail market of São Paulo city were positive for Aeromonas sp, as detected by the direct plating method. When the presence/absence method was used, the positivity was 42%. A. caviae was the most frequent species, followed by A. hydrophila and A. sobria. Production of cytotoxic enterotoxin, observed in suckling mouse assay, was detected in 67% of A. sobria strains, in 60% of A. hydrophila strains and in 40% of A. caviae strains. In vitro tests, performed with HEp-2 cells, showed that 88% of A. hydrophila, 27% of A. sobria and 13% of A. caviae strains were positive for this toxin. The in vivo production of cytotonic enterotoxin, tested after heating the filtrates at 56ºC for 20 minutes, was detected in 17% of A. sobria, in 10% of A. caviae and in none of A. hydrophila strains in vivo. All analyzed strains did not alter HEp-2 cells. 20% and 16% of A. sobria and A. caviae isolates, respectively, presented capacity to adhere to HEp-2 cells. In counterpart, invasion of HEp-2 cells was not observed in any isolate. The Aeromonas isolates were sensitive to the majority of the antimicrobiol agents tested.

In an ex vivo agar plate assay, we monitored the appearance of an inhibitory halo against Vibrio cholerae from the feces of Wistar and Fischer rats aged 10 to 42 days. The frequency of Wistar rats showing halo increased from 0% (10 days) to a maximum of 80.0% (29 days) and then decreased to 53.3% (42 days). A similar pattern was obtained with Fischer rats but with a lower intensity (maximum frequency of 50.0% by day 36). In a separate experiment, when Wistar rats were fed a low-protein diet for 7 days, the inhibitory halo decreased drastically. Three apparently different colony morphologies were isolated from the dominant fecal microbiota: a facultative anaerobe (FAN) and two strict anaerobes (SAN). The ex vivo inhibitory test showed a halo around the feces of germfree mice monoassociated with the FAN bacterium or one of the SAN bacterium but not of the germfree ones. After oral challenge of all groups with V. cholerae, a permissive and a drastic barrier effects were observed in mice with FAN and SAN associated bacteria, respectively. The FAN and one SAN bacteria used in the in vivo challenges were identified as Escherichia coli and Streptococcus intermedius, respectively. The potent antagonism developed by the rat intestinal microbiota against V. cholerae seems to be due, in part, to diffusible compounds and this phenomenon depends apparently on age, strain and nutrition of the animals. These preliminary results also suggest that this effect was due to more than one bacterial component at any given moment.

Molecular evolution in bacteria is examined with an emphasis on the self-assembly of cells capable of primitive division and growth during early molecular evolution. Also, the possibility that some type of encapsulation structure preceeded biochemical pathways and the assembly of genetic material is examined. These aspects will be considered from an evolutionary perspective.

The use of other inducers as substitutes for pectin was studied aiming to reduce the production costs of pectic enzymes. The effects of sugar-cane juice on the production of pectin lyase (PL) and polygalacturonase (PG) by Penicillium griseoroseum were investigated. The fungus was cultured in a mineral medium (pH 6.3) in a rotary shaker (150 rpm) for 48 h at 25oC. Culture media were supplemented with yeast extract and sucrose or sugar-cane juice. Sugar-cane juice added singly to the medium promoted higher PL activity and mycelial dry weight when compared to pectin and the use of sugar-cane juice and yeast extract yielded levels of PG activity that were similar to those obtained with sucrose-yeast extract or pectin. The results indicated that, even at low concentrations, sugar-cane juice was capable of inducing pectin lyase and polygalacturonase with no cellulase activity in P. griseoroseum.

Large amounts of oily sludge are generated as residues by the oil industry, representing a real problem for refineries. This work studied the technical viability of treating oily sludge biologically, through stimulation of native microorganisms, at bench scale. Such microorganisms were able to grow in a medium containing oily sludge as the only carbon and energy sources. Two oily sludge concentrations were studied, 5% (v/v) and 10% (v/v), with a C:N ratio of 100:1. Higher microbial populations were observed in the first case. Substrate inhibition and/or toxic effect took place in the second case. The importance of aeration on the microbial activity and on the biodegradation of the residue was ascertained. In terms of n-paraffins, pristane and phytane consumption, maximum global efficiency of 76.9% (w/w) was achieved, in a medium containing 5% (v/v) of oily sludge. Bacteria of the genus Pseudomonas predominated. Two yeast species were also identified and two filamentous fungi were isolated.

The influence of aeration and automatic pH control on the production of <FONT FACE="Symbol">a</FONT>-amylase by a strain of Bacillus subtilis NRRL 3411 from acid cheese whey was studied. Tests were carried out in a rotary shaker and in mechanically stirred fermenters. <FONT FACE="Symbol">a</FONT>-amylase was analysed according to DUN’s method. Oxygen absorption rate was determined by Cooper’s method. Cell oxygen demand was determined as oxygen consumption in a Warburg respirometer. The level of dissolved oxygen was measured by means of a galvanic silver-lead electrode. Results suggest the possibility of industrial use of acid cheese whey as a carbon source for <FONT FACE="Symbol">a</FONT>-amylase production, since the yield was similar to that produced with lactose. The highest <FONT FACE="Symbol">a</FONT>-amylase levels 10,000 DUN/ml units were not attained at higher aeration rates -431 mLO2/L.h-. The indicated value correspond to a 96 h process with automatic pH control at 7.5. These conditions resulted in double concentration of <FONT FACE="Symbol">a</FONT>-amylase. The enzyme production was directly related to growth in the form of cell aggregates.