Claudia Duarte da Cunha

Oil biodegradation by Bacillus strains isolated from the rock of an oil reservoir located in a deep-water production basin in Brazil

Sixteen spore forming Gram-positive bacteria were isolated from the rock of an oil reservoir located in a deep-water production basin in Brazil. These strains were identified as belonging to the genus Bacillus using classical biochemical techniques and API 50CH kits, and their identity was confirmed by sequencing of part of the 16S rRNA gene. All strains were tested for oil degradation ability in microplates using Arabian Light and Marlin oils and only seven strains showed positive results in both kinds of oils. They were also able to grow in the presence of carbazole, n-hexadecane and polyalphaolefin (PAO), but not in toluene, as the only carbon sources. The production of key enzymes involved with aromatic hydrocarbons biodegradation process by Bacillus strains (catechol 1,2-dioxygenase and catechol 2,3-dioxygenase) was verified spectrophotometrically by detection of cis,cis-muconic acid and 2-hydroxymuconic semialdehyde, and results indicated that the ortho ring cleavage pathway is preferential. Furthermore, polymerase chain reaction (PCR) products were obtained when the DNA of seven Bacillus strains were screened for the presence of catabolic genes encoding alkane monooxygenase, catechol 1,2-dioxygenase, and/or catechol 2,3-dioxygenase. This is the first study on Bacillus strains isolated from an oil reservoir in Brazil.

Gasoline biodegradation in different soil microcosms

The objective of this study was to evaluate gasoline biodegradation in batch soil microcosms. Microorganisms able to grow in the presence of gasoline were isolated from soil. Several treatment systems were performed using both isolated strains and Pseudomonas putida obtained from a culture collection. The treatment system using only autochthonous microflora (system 1) presented an average value of degradation of 50%. The association of Pseudomonas putida, Burkholderia cepacia, Pseudomonas alcaligenes and the native soil microflora (system 13) presented significant percentage of removal of n-undecane (88.7), n-dodecane (61.3) and n-tridecane (66.7). According to these results, systems 1 and 13 revealed considerable potential for application in bioremediation treatments.

Novel Bacterial Phylotypes in Endodontic Infections

Although molecular studies have revealed potential oral pathogens among the phyla Spirochaetes and Deferribacteres, their occurrence in endodontic infections has not been consistently investigated. In this study, we devised a nested PCR-DGGE approach to survey samples from infected root canals for the presence of members of these two phyla, and to examine their diversity. The primers used also amplified DNA from Atopobium species. Eight of 10 cases showed bands representative of the target bacterial groups. DGGE profiles revealed a mean number of 6.5 intense and faint bands. No single band occurred in all profiles. Sequences from intense bands excised from the gel showed similarities to species/phylotypes of all target groups—Flexistipes species (Deferribacteres phylum), uncharacterized spirochetes, and Atopobium species. Analysis of these data indicates that uncultivated Spirochaetes and Deferribacteres phylotypes are frequent members of the endodontic microbiota and may be potential pathogens involved with the etiology of periradicular diseases.

Preliminary identification of the bioremediation limiting factors of a clay bearing soil contaminated with crude oil

Bioremediation is an attractive alternative to treat soils contaminated with petroleum hydrocarbons. However, the effectiveness of biodegradation process can be limited by both contaminant characteristics and its bioavailability in soil. This work aims at establishing a preliminary procedure to identify the main factor (hydrocarbon recalcitrance or its bioavailability) that impairs the biodegradation, possibly resulting in low remediation efficiencies. Tests in soil microcosms were carried out applying biostimulation and bioaugmentation techniques. Similar experiments were conducted using liquid medium containing crude oil and microorganisms, both of them extracted from a weathered contaminated soil sample, nutrients and microbial inoculum constituted of Rhodoturula glutinis and Nocardia nova. After 30 days, highest microcosms biodegradation levels (98.8%) were obtained with bioaugmentation in liquid medium, pointing out that the main problem of petroleum hydrocarbon utilization by microorganisms was associated to its availability and not only to the intrinsic recalcitrancy of remaining pollutants.