Brazilian Journal of Microbiology, Volume: 41, Issue: 4, Published: 2010
  • Conventional and alternative antifungal therapies to oral candidiasis Review

    Anibal, Paula Cristina; Sardi, Janaina de Cássia Orlandi; Peixoto, Iza Teixeira Alves; Moraes, Julianna Joanna de Carvalho; Höfling, José Francisco

    Abstract in English:

    Candida-associated denture stomatitis is the most common form of oral candidal infection, with Candida albicans being the principal etiological agent. Candida adheres directly or via an intermediary layer of plaque-forming bacteria to denture acrylic. Despite antifungal therapy to treat denture stomatitis, infection is reestablished soon after the treatment ceases. In addition, many predisposing factors have been identified as important in the development of oral candidiasis, including malnourishment, common endocrine disorders, such as diabetis mellitus, antibacterial drug therapy, corticosteroids, radiotherapy and other immunocompromised conditions, such as acquired immunodeficiency syndrome (AIDS). These often results in increased tolerance to the most commonly used antifungals. So this review suggests new therapies to oral candidiasis.
  • The role of mycorrhization helper bacteria in the establishment and action of ectomycorrhizae associations Review

    Rigamonte, Tatiana Alves; Pylro, Victor Satler; Duarte, Gabriela Frois

    Abstract in English:

    More than 95 % short roots of most terrestrial plants are colonized by mycorrhizal fungi as soon as they emerge in the upper soil profiles. The establishment of mycorrhizal association involves profound morphological and physiological changes in root and fungus. It is affected by other rhizospheric microorganisms, specifically by the bacteria. Bacteria may have developed mechanisms of selective interaction with surrounding microorganisms, with neutral or positive effects on mycorrhizal associations, but negative effect on root pathogens in general. Because of the beneficial effect of bacteria on mycorrhizae, the concept of Mycorrhization Helper Bacteria (MHB) was created. Five main actions of MHB on mycorrhizae were proposed: in the receptivity of root to the mycobiont, in root-fungus recognition, in fungal growth, in the modification of rhizospheric soil and in the germination of fungal propagules. MHB appear to develop a gradation of specificity for the mycobiont, but little or no specificity for the host plant in symbiosis. One of the main groups of MHB is the fluorescent Pseudomonas, well represented in diversity and cell density studies of mycorrhizal associations. This review covers the activity of MHB in the establishment of ectomycorrhizae, taking as model the effects of Pseudomonas sp. described in scientific literature.
  • Candida albicans: genotyping methods and clade related phenotypic characteristics Review

    Lyon, Juliana P.; Moraes, Karen C.M.; Moreira, Leonardo M.; Aimbire, Flávio; Resende, Maria Aparecida de

    Abstract in English:

    Several molecular methods, such as Southern blotting hybridization, Multilocus Sequence Typing, and DNA microsatellite analysis, have been employed to genotype Candida albicans. The genotype analysis allows to group strains in clades, that is, a group composed of one ancestor and its descendants. These genotype studies demonstrate that clades distribution is influenced by geographic area as well as that antifungal resistance is associated with particular clades. These findings suggested that C. albicans reproduces mainly in a clonal manner, with certain degree of DNA microevolution. Additionally, virulence factors and site of isolation have also been associated with clade specificity. The present article is a brief review about the methods used for Candida genotyping and the correlated clade systems established. Special emphasis is given to Ca3 hybridization, MLST, and Microsatellites. The present work is also focused on the phenotypic and physiological traits associated with Candida clades.
  • Application of microbial α-amylase in industry - A review Review

    Souza, Paula Monteiro de; Magalhães, Pérola de Oliveira e

    Abstract in English:

    Amylases are one of the main enzymes used in industry. Such enzymes hydrolyze the starch molecules into polymers composed of glucose units. Amylases have potential application in a wide number of industrial processes such as food, fermentation and pharmaceutical industries. α-Amylases can be obtained from plants, animals and microorganisms. However, enzymes from fungal and bacterial sources have dominated applications in industrial sectors. The production of α-amylase is essential for conversion of starches into oligosaccharides. Starch is an important constituent of the human diet and is a major storage product of many economically important crops such as wheat, rice, maize, tapioca, and potato. Starch-converting enzymes are used in the production of maltodextrin, modified starches, or glucose and fructose syrups. A large number of microbial α-amylases has applications in different industrial sectors such as food, textile, paper and detergent industries. The production of α-amylases has generally been carried out using submerged fermentation, but solid state fermentation systems appear as a promising technology. The properties of each α-amylase such as thermostability, pH profile, pH stability, and Ca-independency are important in the development of fermentation process. This review focuses on the production of bacterial and fungal α-amylases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications.
  • Biotechnological production of citric acid Review

    Max, Belén; Salgado, José Manuel; Rodríguez, Noelia; Cortés, Sandra; Converti, Attilio; Domínguez, José Manuel

    Abstract in English:

    This work provides a review about the biotechnological production of citric acid starting from the physicochemical properties and industrial applications, mainly in the food and pharmaceutical sectors. Several factors affecting citric acid fermentation are discussed, including carbon source, nitrogen and phosphate limitations, pH of culture medium, aeration, trace elements and morphology of the fungus. Special attention is paid to the fundamentals of biochemistry and accumulation of citric acid. Technologies employed at industrial scale such as surface or submerged cultures, mainly employing Aspergillus niger, and processes carried out with Yarrowia lipolytica, as well as the technology for recovering the product are also described. Finally, this review summarizes the use of orange peels and other by-products as feedstocks for the bioproduction of citric acid.
  • Molecular cloning, characterization and enzymatic properties of a novel βeta-agarase from a marine isolate Psudoalteromonas sp. AG52 Environmental Microbiology

    Oh, Chulhong; Nikapitiya, Chamilani; Lee, Youngdeuk; Whang, Ilson; Kang, Do-Hyung; Heo, Soo-Jin; Choi, Young-Ung; Lee, Jehee

    Abstract in English:

    An agar-degrading Pseudoalteromonas sp. AG52 bacterial strain was identified from the red seaweed Gelidium amansii collected from Jeju Island, Korea. A β-agarase gene which has 96.8% nucleotide identity to Aeromonas β-agarase was cloned from this strain, and was designated as agaA. The coding region is 870 bp, encoding 290 amino acids and possesses characteristic features of the glycoside hydrolase family (GHF)-16. The predicted molecular mass of the mature protein was 32 kDa. The recombinant β-agarase (rAgaA) was overexpressed in Escherichia coli and purified as a fusion protein. The optimal temperature and pH for activity were 55 ºC and 5.5, respectively. The enzyme had a specific activity of 105.1 and 79.5 unit/mg toward agar and agarose, respectively. The pattern of agar hydrolysis demonstrated that the enzyme is an endo-type β-agarase, producing neoagarohexaose and neoagarotetraose as the final main products. Since, Pseudoalteromonas sp. AG52 encodes an agaA gene, which has greater identity to Aeromonas β-agarase, the enzyme could be considered as novel, with its unique bio chemical characteristics. Altogether, the purified rAgaA has potential for use in industrial applications such as development of cosmetics and pharmaceuticals.
  • Distribution of prokaryotic organisms in a tropical estuary influenced by sugar cane agriculture in northeast Brazil Environmental Microbiology

    Wolf, Lars; Schwalger, Berit; Knoppers, Bastiaan A.; Silva, Luiz Antonio Ferreira da; Medeiros, Paulo Ricardo Petter; Pollehne, Falk

    Abstract in English:

    In a joint Brazilian-German case study, distribution patterns of microorganisms were compared with environmental variables in the tropical coastal Manguaba lagoon in northeast Brazil, which is situated downstream of several sugar cane processing plants . 16S rDNA and 16S rRNA single strand conformation polymorphism (SSCP) gene fingerprinting were used to follow the composition and distribution of microorganisms throughout the salinity gradient of the lagoon. Potentially abundant microorganisms were identified by sequencing representative SSCP bands. It could be demonstrated that the distribution of microbes was in close relation to the physico-chemical environmental settings and followed a common scheme. In the in- and outlet areas of the lagoon rather transient microbial communities were found, whereas in the central part a stable, diverse community was encountered, that due to the long residence time of the water, had ample time for development and adaptation.
  • Characterization of Bacillus isolates of potato rhizosphere from andean soils of Peru and their potential PGPR characteristics Environmental Microbiology

    Calvo, Pamela; Ormeño-Orrillo, Ernesto; Martínez-Romero, Esperanza; Zúñiga, Doris

    Abstract in English:

    Bacillus spp. are well known rhizosphere residents of many crops and usually show plant growth promoting (PGP) activities that include biocontrol capacity against some phytopatogenic fungi. Potato crops in the Andean Highlands of Peru face many nutritional and phytophatogenic problems that have a significant impact on production. In this context is important to investigate the natural presence of these microorganisms in the potato rhizosphere and propose a selective screening to find promising PGP strains. In this study, sixty three Bacillus strains isolated from the rhizosphere of native potato varieties growing in the Andean highlands of Peru were screened for in vitro antagonism against Rhizoctonia solani and Fusarium solani. A high prevalence (68%) of antagonists against R. solani was found. Ninety one percent of those strains also inhibited the growth of F. solani. The antagonistic strains were also tested for other plant growth promotion activities. Eighty one percent produced some level of the auxin indole-3-acetic acid, and 58% solubilized tricalcium phosphate. Phylogenetic analysis revealed that the majority of the strains belonged to the B. amyloliquefaciens species, while strains Bac17M11, Bac20M1 and Bac20M2 may correspond to a putative new Bacillus species. The results suggested that the rhizosphere of native potatoes growing in their natural habitat in the Andes is a rich source of Bacillus fungal antagonists, which have a potential to be used in the future as PGP inoculants to improve potato crop.
  • Role of brown-rot fungi in the bioremoval of azo dyes under different conditions Environmental Microbiology

    Ali, Naeem; Hameed, Abdul; Ahmed, Safia

    Abstract in English:

    The present study is vital to the understanding of bioremediation of structurally different azo dyes by some unusual Brown-rot fungi. Bioremoval of each dye (20 mg l-1) was tested in two different culture media under static and shaking conditions by taking inocula from different fungi. Fungal strains showed varying dyes removal abilities, though considerable high in case of Acid Red (AR) 151(di-azo) as compared to Orange (Or) II (mono-azo). With an exception of Aspergillus tereus SA3, all the fungal isolates showed higher removal of dyes in SDB. Under static condition, the maximum decolorizing fungal strains were; Aspergillus flavus SA2 (67%) and Alternaria spp. SA4 (57%) in AR 151, while Penicillium spp. (34 and 33 %) in Orange II, in SDB and STE, respectively. Bioremoval of dyes was considerably increased when experiments were shifted from static to shaking mode. It was specifically increased (%) in; AR 151 (255) with Penicillium spp., Or II with A. flavus SA2 (112) and Alternaria spp. (111). The primary mechanism of dyes removal proved to be fungal biosorption. However, reduction of dyes (onto fungal) with formation of their products (α. naphthol, sulphalinic acid and aniline) furthermore revealed that dyes (specifically azo) were actually biodegraded.
  • Isolation and characteristics of lactic acid bacteria isolated from ripe mulberries in Taiwan Environmental Microbiology

    Chen, Yi-sheng; Wu, Hui-chung; Yanagida, Fujitoshi

    Abstract in English:

    The objective of this study was to isolate, characterize, and identify lactic acid bacteria (LAB) from ripe mulberries collected in Taiwan. Ripe mulberry samples were collected at five mulberry farms, located in different counties of Taiwan. Eighty-eight acid-producing cultures were isolated from these samples, and isolates were divided into classes first by phenotype, then into groups by restriction fragment length polymorphism (RFLP) analysis and sequencing of 16S ribosomal DNA (rDNA). Phenotypic and biochemical characteristics led to identification of four bacterial groups (A to D). Weissella cibaria was the most abundant type of LAB distributed in four mulberry farms, and Lactobacillus plantarum was the most abundant LAB found in the remaining farm. Ten W. cibaria and one Lactococcus lactis subsp. lactis isolate produced bacteriocins against the indicator strain Lactobacillus sakei JCM 1157T. These results suggest that various LAB are distributed in ripe mulberries and W. cibaria was the most abundant LAB found in this study.
  • Biodegradation of naphthalene and anthracene by chemo-tactically active rhizobacteria of populus deltoides Environmental Microbiology

    Bisht, Sandeep; Pandey, Piyush; Sood, Anchal; Sharma, Shivesh; Bisht, N. S.

    Abstract in English:

    Several naphthalene and anthracene degrading bacteria were isolated from rhizosphere of Populus deltoides, which were growing in non-contaminated soil. Among these, four isolates, i.e. Kurthia sp., Micrococcus varians, Deinococcus radiodurans and Bacillus circulans utilized chrysene, benzene, toluene and xylene, in addition to anthracene and naphthalene. Kurthia sp and B. circulans showed positive chemotactic response for naphthalene and anthracene. The mean growth rate constant (K) of isolates were found to increase with successive increase in substrate concentration (0.5 to 1.0 mg/50ml). B. circulans SBA12 and Kurthia SBA4 degraded 87.5% and 86.6% of anthracene while, Kurthia sp. SBA4, B. circulans SBA12, and M. varians SBA8 degraded 85.3 %, 95.8 % and 86.8 % of naphthalene respectively after 6 days of incubation as determined by HPLC analysis.
  • Degradation of h-acid by free and immobilized cells of Alcaligenes latus Environmental Microbiology

    Usha, M.S.; Sanjay, M.K.; Gaddad, S.M.; Shivannavar, C.T.

    Abstract in English:

    Alcaligenes latus, isolated from industrial effluent, was able to grow in mineral salts medium with 50 ppm (0.15 mM) of H-acid as a sole source of carbon. Immobilization of Alcaligenes latus in Ca-alginate and polyurethane foam resulted in cells embedded in the matrices. When free cells and immobilized cells were used for biodegradation studies at concentration ranging from 100 ppm (0.3 mM) to 500 ppm (1.15 mM) degradation rate was enhanced with immobilized cells. Cells immobilized in polyurethane foam showed 100% degradation up to 350 ppm (1.05 mM) and 57% degradation at 500 ppm (1.5 mM). Degradation rate of Ca-alginate immobilized cells was less as compared to that of polyurethane foam immobilized cells. With Ca-alginate immobilized cells 100% degradation was recorded up to 200 ppm (0.6 mM) of H-acid and only 33% degradation was recorded at 500 ppm (1.5 mM) of H-acid. Spectral analysis of the products after H-acid utilization showed that the spent medium did not contain any aromatic compounds indicating H-acid degradation by A. latus.
  • Comparative study of wild and transformed salt tolerant bacterial strains on Triticum aestivum growth under salt stress Environmental Microbiology

    Afrasayab, Shazia; Faisal, Muhammad; Hasnain, Shahida

    Abstract in English:

    Eleven salt tolerant bacteria isolated from different sources (soil, plants) and their transformed strains were used to study their influence on Triticum aestivum var. Inqlab-91 growth under salt (100 mM NaCl) stress. Salt stress caused reduction in germination (19.4%), seedling growth (46%) and fresh weight (39%) in non-inoculated plants. In general, both wild and transformed strains stimulated germination, seedling growth and fresh weight in salt free and salt stressed conditions. At 100 mM NaCl, Staphylococcus xylosus ST-1 caused 25% increments in seedling length over respective control. Soluble protein content significantly enhanced (49%) under salt stress as compared to salt free control. At 100 mM NaCl parental strain PT-5 resulted about 32% enhancement in protein content over respective control treatment. Salt stress induced the promotion of auxin content in seedlings. Overall, Bacillus subtilis HAa2 and transformed E. coli-SP-7-T, caused 33% and 30% increases in auxin content, respectively, were recorded under salt stress in comparison to control.
  • The effect of different growth regimes on the endophytic bacterial communities of the fern, Dicksonia sellowiana hook (Dicksoniaceae) Environmental Microbiology

    Barros, Irene de Araújo; Araújo, Welington Luiz; Azevedo, João Lúcio

    Abstract in English:

    Endophytic bacteria associated with the fern Dicksonia sellowiana were investigated. The bacterial communities from the surface-sterilized pinnae and rachis segments of the plants from the Brazilian Atlantic Rainforest that grew in native field conditions were compared with the bacterial communities from plants grown in greenhouses and plants that were initially grown in greenhouses and then transferred to the forest. From 540 pinnae and 540 rachis segments, 163 (30.2%) and 346 (64.2%) were colonized by bacteria, respectively. The main bacterial genera and species that were isolated included Bacillus spp. (B. cereus, B. megaterium, B. pumilus and B. subtilis), Paenibacillus sp., Amphibacillus sp., Gracilibacillus sp., Micrococcus sp. and Stenotrophomonas spp. (S. maltophilia and S. nitroreducens). B. pumilus was the most frequently isolated bacterial species. Amphibacillus and Gracilibacillus were reported as endophytes for the first time. Other commonly found bacterial genera were not observed in D. sellowiana, which may reflect preferences of specific bacterial communities inside this fern or detection limitations due to the isolation procedures. Plants that were grown in greenhouses and plants that were reintroduced into the forest displayed more bacterial genera and species diversity than native field plants, suggesting that reintroduction shifts the bacterial diversity. Endophytic bacteria that displayed antagonistic properties against different microorganisms were detected, but no obvious correlation was found between their frequencies with plant tissues or with plants from different growth regimes. This paper reports the first isolation of endophytic bacteria from a fern.
  • Antimicrobial resistance and PCR-ribotyping of Shigella responsible for foodborne outbreaks occurred in southern Brazil Food Microbiology

    Paula, Cheila Minéia Daniel de; Geimba, Mercedes Passos; Amaral, Patrícia Heidrich do; Tondo, Eduardo Cesar

    Abstract in English:

    Little information about Shigella responsible for foodborne shigellosis is available in Brazil. The present study aimed to investigate the antimicrobial resistance and PCR-ribotyping patterns of Shigella isolates responsible for foodborne outbreaks occurred in Rio Grande do Sul State (RS), Southern Brazil in the period between 2003 and 2007. Shigella strains (n=152) were isolated from foods and fecal samples of victims of shigellosis outbreaks investigated by the Surveillance Service. Identification of the strains at specie level indicated that 71.1% of them were S. flexneri, 21.5% S. sonnei, and 0.7% S. dysenteriae. Ten strains (6.7%) were identified only as Shigella spp. An increasing occurrence of S. sonnei was observed after 2004. Most of the strains were resistant to streptomycin (88.6%), followed by ampicillin (84.6%), and sulfamethoxazole/trimethoprim (80.5 %). Resistant strains belonged to 73 patterns, and pattern A (resistance to ampicillin, sulfamethoxazole/trimethoprim, tetracycline, streptomycin, chloramphenicol, and intermediate resistance to kanamycin) grouped the largest number of isolates (n=36). PCR-ribotyping identified three banding patterns (SH1, SH2, and SH3). SH1 grouped all S. flexneri and SH2 grouped all S. sonnei. The S. dysenteriae strain belonged to group SH3. According to the results, several Shigella isolates shared the same PCR-rybotyping banding pattern and the same resistance profile, suggesting that closely related strains were responsible for the outbreaks. However, other molecular typing methods need to be applied to confirm the clonal relationship of these isolates.
  • Virulence and cytotoxicity of seafood borne Aeromonas hydrophila Food Microbiology

    Illanchezian, Seethalakshmi; Jayaraman, SathishKumar; Manoharan, Muthu Saravanan; Valsalam, Saritha

    Abstract in English:

    The present study was conducted to determine the virulence and cytotoxicity of Aeromonas hydrophila strains isolated from seafood samples collected from 5 major fish markets in Chennai, Tamil Nadu, India. Among 73 A. hydrophila strains isolated from fish and shrimp samples, 86.3% exhibited haemolysis, 78.1% produced slime, 98.63% produced protease and also demonstrated cytotoxicity on Vero cells. Cell shrinkage, detachment and rounding of Vero cells were recorded as cytotoxic changes. Only one strain did not show haemolysis, slime production, proteolytic activity and cytotoxicity on treatment with Vero cells. Positive correlation was observed between proteolytic activity and cytotoxicity irrespective of haemolytic activity of the strains. These results demonstrated the presence of wide spread, pathogenically characterized, cytotoxic seafood borne A. hydrophila in Chennai.
  • Assessment of hydrophobicity and roughness of stainless steel adhered by an isolate of Bacillus cereus from a dairy plant Food Microbiology

    Bernardes, Patrícia Campos; Andrade, Nélio José de; Ferreira, Sukarno Olavo; Sá, João Paulo Natalino de; Araújo, Emiliane Andrade; Delatorre, Deyse Maria Zanom; Luiz, Lívia Maria Pinheiro

    Abstract in English:

    The interaction between the surface of stainless steel and Bacillus cereus was studied in terms of the characteristics of interfacial interaction determined from the measurement of the contact angle of the surface of B. cereus and stainless steel in the presence or absence of B. cereus adherence. The microtopographies and the roughness of the surface of stainless steel and stainless steel adhered by B. cereus were evaluated with the help of atomic force microscopy and perfilometry. The strain of B. cereus studied was considered hydrophilic, whereas the stainless steel was considered hydrophobic. The adhesion was not thermodynamically favorable (ΔGadhesion > 0) between the stainless steel and the strain of B. cereus studied. Thus, the interaction between them was not favored by the thermodynamic aspect of adhesion. There was no difference (p > 0.05) in the roughness of the surfaces of stainless steel adhered by B. cereus when analyzed by atomic force microscope and perfilometry.
  • Detection of shigella in lettuce by the use of a rapid molecular assay with increased sensitivity Food Microbiology

    Jiménez, Kenia Barrantes; McCoy, Clyde B.; Achí, Rosario

    Abstract in English:

    A Multiplex Polymerase Chain Reaction (PCR) assay to be used as an alternative to the conventional culture method in detecting Shigella and enteroinvasive Escherichia coli (EIEC) virulence genes ipaH and ial in lettuce was developed. Efficacy and rapidity of the molecular method were determined as compared to the conventional culture. Lettuce samples were inoculated with different Shigella flexneri concentrations (from 10 CFU/ml to 10(7) CFU/ml). DNA was extracted directly from lettuce after inoculation (direct-PCR) and after an enrichment step (enrichment PCR). Multiplex PCR detection limit was 10(4) CFU/ml, diagnostic sensitivity and specificity were 100% accurate. An internal amplification control (IAC) of 100 bp was used in order to avoid false negative results. This method produced results in 1 to 2 days while the conventional culture method required 5 to 6 days. Also, the culture method detection limit was 10(6) CFU/ml, diagnostic sensitivity was 53% and diagnostic specificity was 100%. In this study a Multiplex PCR method for detection of virulence genes in Shigella and EIEC was shown to be effective in terms of diagnostic sensitivity, detection limit and amount of time as compared to Shigella conventional culture.
  • Incorporation of nisin in natural casing for the control of spoilage microorganisms in vacuum packaged sausage Food Microbiology

    Barros, Joyce Regina de; Kunigk, Leo; Jurkiewicz, Cynthia Hyppolito

    Abstract in English:

    This study aimed to evaluate the effectiveness of natural casing treatment with nisin and phosphoric acid on control of spoilage microorganisms in vacuum packaged sausages. Ovine casings were dipped in the following baths: 1) 0.1% food grade phosphoric acid; 2) 5.0 mg/L nisin; 3) 0.1% phosphoric acid and 5.0 mg/L nisin; and 4) sterile water (control). The sausages were produced in a pilot plant, stuffed into the pretreated natural casings, vacuum packaged and stored at 4 and 10 °C for 56 days. The experiments were performed according to a full factorial design 2³, totalizing 8 treatments that were repeated in 3 blocks. Aerobic plate counts and lactic acid bacteria analysis were conducted at 1, 14, 28, 42 and 56 days of storage. Treatment of casings with phosphoric acid 0.1% alone did not inhibit the growth of lactic acid bacteria and reduced the aerobic plate count by 1 log. The activity of nisin against lactic acid bacteria was enhanced by the addition of phosphoric acid, demonstrating a synergistic effect. Furthermore nisin activity was more evident at lower storage temperature (4 ºC). Therefore treatment of the natural casings with nisin and phosphoric acid, combined with low storage temperature, are obstacles that present a potential for controlling the growth of lactic acid bacteria in vacuum packaged sausage.
  • Production of bacteriocin-like substances by lactic acid bacteria isolated from regional ovine cheese Food Microbiology

    Nespolo, Cássia Regina; Brandelli, Adriano

    Abstract in English:

    Lactic acid bacteria (LAB) were isolated from ovine milk and cheeses manufactured in the South Region of Brazil. Among 112 bacterial isolates investigated, 59 were chosen through a screening for LAB. Among these 59 strains of LAB, 21% showed antimicrobial, proteolytic and lipolytic activities. Based on this screening, Lactobacillus plantarum LCN 17 and Lactobacillus rhamnosus LCN 43 were selected and tested for the production of bacteriocin-like substances (BLS). The BLS produced by both isolates showed antimicrobial activity against Listeria monocytogenes, whereas that produced by L. plantarum LCN 17 presented higher stability to different temperature, pH and enzyme treatments. These strains present potential for production of BLS, and for use as starter cultures.
  • Inhibition of mycotoxin-producing Aspergillus nomius vsc 23 by lactic acid bacteria and Saccharomyces cerevisiae Food Microbiology

    Muñoz, R; Arena, M.E.; Silva, J; González, S.N.

    Abstract in English:

    The effect of different fermenting microorganisms on growth of a mycotoxin- producing Aspergillus nomius was assayed. Two lactic acid bacteria, Lactobacillus fermentum and Lactobacillus rhamnosus, and Saccharomyces cerevisiae, all of which are widely used in fermentation and preservation of food, were assayed on their fungus inhibitory properties. Assays were carried out by simultaneous inoculation of one of the possible inhibiting microorganisms and the fungus or subsequent inoculation of one of the microorganisms followed by the fungus. All three microorganisms assayed showed growth inhibition of the mycotoxin-producing Aspergillus strain. L. rhamnosus O236, isolated from sheep milk and selected for its technological properties, showed highest fungal inhibition of the microorganisms assayed. The use of antifungal LAB with excellent technological properties rather than chemical preservatives would enable the food industry to produce organic food without addition of chemical substances.
  • Adhesion and biocides inactivation of Salmonella on stainless steel and polyethylene Food Microbiology

    Tondo, Eduardo Cesar; Machado, Taís Raquel Marcon; Malheiros, Patrícia da Silva; Padrão, Débora Kruger; Carvalho, Ana Lyl de; Brandelli, Adriano

    Abstract in English:

    The adhesion of Salmonella (S.) strains to stainless steel and polyethylene and their inactivation by biocides used in food industry was investigated. Coupons of stainless steel and polyethylene were immersed in bacterial suspensions of S. Enteritidis, S. Typhimurium, and S. Bredeney during 15, 30, and 60 minutes, and submitted to different concentrations of peracetic acid (PAA), sodium hypochlorite (NaOCl), and quaternary ammonium (Quat) sanitizers. Hydrophobicity of the surfaces was evaluated by contact angle measurements using the sessile drop method and bacterial adhesion was accompanied through bacterial counts and scanning electron microscopy (SEM). Results indicated that the three serovars of Salmonella presented similar adhesion to both materials (5.0 to 6.5 log cfu cm-2). The time of exposure did not influence the counts of adhered cells on both surfaces, however SEM revealed larger clusters of S. Enteritidis on both materials, not found for the other serovars. S. Enteritidis presented lower sessile drop angle on polyethylene, indicating hydrophilic properties of this material. The biocides were not able to inactivate all the microorganisms adhered on both surfaces. At least 1 log cfu cm-2 of all serovars tested remained viable after the exposure to different biocide concentrations. In general, higher counts of survivors were observed on polyethylene disinfected with different concentrations of biocides. S. Bredeney e S. Typhimurium were more resistant than S. Enteritidis to PAA, whilst S. Enteritidis presented smaller reduction rates to NaOCl. This last biocide was able to reduce Salmonella counts in approximately 3.0 to 4.0 log cm-2. When adhered to polyethylene, the serovars S. Typhimurium and S. Enteritidis were more resistant to Quat than S. Bredeney in all concentrations tested, and the numbers of S. Enteritidis remained almost unaltered. On stainless steel disinfected by Quat, S. Bredeney presented higher numbers of survivors.
  • Evaluation of different cryoprotective agents in maintenance of viability of Helicobacter pylori in stock culture media Medical Microbiology

    Oskouei, Daryoush Davoudi; Bekmen, Neslihan; Ellidokuz, Hülya; Yilmaz, Özlem

    Abstract in English:

    Four different cryoprotective supplemented stock media were evaluated for maintaining better survival and recovery of H. pylori type strain NCTC 11637 at two different maintenance temperatures of -20°C and -80°C after one month preservation as frozen stocks. The spread plate colony count method was used to investigate the recovery rate of H. pylori from equally inoculated bacterial suspensions in differently prepared stock cultures. After the preservation of H. pylori for one month in different cryoprotectant-supplemented stock media, the recovery rates for -20°C obtained for stock cultures supplemented with dimethyl sulfoxide (DMSO), polyethylene glycol (PEG), glycerol and glycerol+sucrose, as well as controls with and without human serum alone were 7.13, 6.97, 7.93, 7.99, 6.95 and 0.0 log CFU/ml, respectively. Maintenance of bacteria at -80°C gave statistically higher recovery rates compared to preservation at -20°C with the values of 8.55, 8.24, 8.59, 8.66, 8.01 and 0.0 log CFU/ml for these above mentioned stock cultures. The stock cultures supplemented with glycerol+sucrose and glycerol showed the highest recovery rates, 7.99 and 7.93 for -20°C vs. 8.66 and 8.59 for -80°C respectively, which were statistically different from the others. Our study revealed that H. pylori type strain NCTC 11637 could be better preserved at -80°C than -20°C. The best stock media which supported viability or culturability of bacteria were brain heart infusion broth (BHI)+glycerol+human serum and BHI+glycerol+sucrose+human serum, where the latter yielded the higher recovery rate.
  • Detection of Group B Streptococcus in Brazilian pregnant women and antimicrobial susceptibility patterns Medical Microbiology

    Castellano-Filho, Didier Silveira; Silva, Vânia Lúcia da; Nascimento, Thiago César; Vieira, Marcel de Toledo; Diniz, Cláudio Galuppo

    Abstract in English:

    Group B Streptococcus (GBS) is still not routinely screened during pregnancy in Brazil, being prophylaxis and empirical treatment based on identification of risk groups. This study aimed to investigate GBS prevalence in Brazilian pregnant women by culture or polymerase chain reaction (PCR) associated to the enrichment culture, and to determine the antimicrobial susceptibility patterns of isolated bacteria, so as to support public health policies and empirical prophylaxis. After an epidemiological survey, vaginal and anorectal specimens were collected from 221 consenting laboring women. Each sample was submitted to enrichment culture and sheep blood agar was used to isolate suggestive GBS. Alternatively, specific PCR was performed from enrichment cultures. Antimicrobial susceptibility patterns were determined for isolated bacteria by agar diffusion method. No risk groups were identified. Considering the culture-based methodology, GBS was detected in 9.5% of the donors. Twenty five bacterial strains were isolated and identified. Through the culture-PCR methodology, GBS was detected in 32.6% specimens. Bacterial resistance was not detected against ampicillin, cephazolin, vancomycin and ciprofloxacin, whereas 22.7% were resistant to erythromycin and 50% were resistant to clindamycin. GBS detection may be improved by the association of PCR and enrichment culture. Considering that colony selection in agar plates may be laboring and technician-dependent, it may not reflect the real prevalence of streptococci. As in Brazil prevention strategies to reduce the GBS associated diseases have not been adopted, prospective studies are needed to anchor public health policies especially considering the regional GBS antimicrobial susceptibility patterns.
  • Microbiota associated with chronic osteomyelitis of the jaws Medical Microbiology

    Gaetti-Jardim Júnior, Elerson; Fardin, Angélica Cristiane; Gaetti-Jardim, Ellen Cristina; Castro, Alvimar Lima de; Schweitzer, Christiane Marie; Avila-Campos, Mario Julio

    Abstract in English:

    Chronic osteomyelitis of maxilla and mandible is rare in industrialized countries and its occurrence in developing countries is associated with trauma and surgery, and its microbial etiology has not been studied thoroughly. The aim of this investigation was to evaluate the microbiota associated with osteomyelitis of mandible or maxilla from some Brazilian patients. After clinical and radiographic evaluation, samples of bone sequestra, purulent secretion, and biopsies of granulomatous tissues from twenty-two patients with chronic osteomyelitis of mandible and maxilla were cultivated and submitted for pathogen detection by using a PCR method. Each patient harbored a single lesion. Bacterial isolation was performed on fastidious anaerobe agar supplemented with hemin, menadione and horse blood for anaerobes; and on tryptic soy agar supplemented with yeast extract and horse blood for facultative bacteria and aerobes. Plates were incubated in anaerobiosis and aerobiosis, at 37ºC for 14 and 3 days, respectively. Bacteria were cultivated from twelve patient samples; and genera Actinomyces, Fusobacterium, Parvimonas, and Staphylococcus were the most frequent. By PCR, bacterial DNA was detected from sixteen patient samples. The results suggest that cases of chronic osteomyelitis of the jaws are usually mixed anaerobic infections, reinforcing the concept that osteomyelitis of the jaws are mainly related to microorganisms from the oral environment, and periapical and periodontal infections may act as predisposing factors.
  • Ethambutol resistance of indigenous Mycobacterium tuberculosis isolated from human patients Medical Microbiology

    Nazir, Taha; Rasool, Muhammad Hidayat; Hameed, Abdul; Ahmad, Bashir; Qureshi, Javed Anver

    Abstract in English:

    The present study was conducted to find out the ethambutol resistance pattern of indigenous isolates of Mycobacterium tuberculosis from Tuberculosis diagnosed human patients. A total of 172 specimens were collected from six different sources and comprised of 84.9% sputum, 10.5% pus and 4.7% bronchial washings. There were 70.9% males and 29.1% females with 84.30% pulmonary and 15.69% extra-pulmonary tuberculosis. The Mycobacterium tuberculosis isolates collected from primary culture were further studied to determine their pattern and level of resistance. The inoculums were prepared using 0.5 Mac Farland turbidity standards. Five different concentration of ethambutol were used in Lowenstein Jensen (LJ) medium i.e. 2μg/ml, 4μg/ml, 6μg/ml, 8μg/ml and 10μg/ml for sensitivity testing. Data showed 10 (5.8%) resistant and 162 (94.2%) sensitive Mycobacterium tuberculosis out of total 172 clinical isolates. The growth was not inhibited at 1st (2μg/ml) and 2nd (4μg/ml) drug levels, while growth of 50% isolates inhibited at 3rd level (6μg/ml), 30% inhibited at 4th level (8μg/ml) and 20% at 5th level (10μg/ml). The last three levels are above the therapeutic index and not recommended in actual clinical practice. It is thus conceivable to explore some other more effective chemotherapeutic agents, modify combinations or find more effective procedures to stop morbidity and mortality due to ethambutol resistant Mycobacterium tuberculosis.
  • Rosmarinus officinalis essential oil: antiproliferative, antioxidant and antibacterial activities Medical Microbiology

    Hussain, Abdullah Ijaz; Anwar, Farooq; Chatha, Shahzad Ali Shahid; Jabbar, Abdul; Mahboob, Shahid; Nigam, Poonam Singh

    Abstract in English:

    The aim of this work was to investigate and compare the antiproliferative, antioxidant and antibacterial activities of Rosmarinus officinalis essential oil, native to Pakistan. The essential oil content from the leaves of R. officinalis was 0.93 g 100g-1. The GC and GC-MS analysis revealed that the major components determined in R. officinalis essential oil were 1,8-cineol (38.5%), camphor (17.1%), α-pinene (12.3%), limonene (6.23%), camphene (6.00%) and linalool (5.70%). The antiproliferative activity was tested against two cancer (MCF-7 and LNCaP) and one fibroblast cell line (NIH-3T3) using the MTT assay, while, the antioxidant activity was evaluated by the reduction of 2, 2-diphenyl-1-picryl hydrazyl (DPPH) and measuring percent inhibition of peroxidation in linoleic acid system. The disc diffusion and modified resazurin microtitre-plate assays were used to evaluate the inhibition zones (IZ) and minimum inhibitory concentration (MIC) of R. officinalis essential oil, respectively. It is concluded from the results that Rosmarinus officinalis essential oil exhibited antiproliferative, antioxidant and antibacterial activities.
  • Comparison of the worldwide transmissible Pseudomonas aeruginosa with isolates from brazilian cystic fibrosis patients Medical Microbiology

    Leão, Robson Souza; Carvalho-Assef, Ana Paula D; Ferreira, Alex Guerra; Folescu, Tânia Wrobel; Barth, Afonso Luís; Pitt, Tyrone Leslie; Marques, Elizabeth Andrade

    Abstract in English:

    Cross-infection with Pseudomonas aeruginosa among cystic fibrosis (CF) patients is a rare occurrence. However, the emergence of transmissible strains has been reported between unrelated individuals. We analyzed the genetic relationship among P. aeruginosa isolates from Brazilian CF patients and transmissible clones which are worldwide spread. The data does not indicate the presence of closely related variant clones.
  • Quantification of biofilm production on polystyrene by Listeria, Escherichia coli and Staphylococcus aureus isolated from a poultry slaughterhouse Veterinary Microbiology

    Rodrigues, Laura Beatriz; Santos, Luciana Ruschel dos; Tagliari, Vinícius Zancanaro; Rizzo, Natalie Nadin; Trenhago, Graciela; Oliveira, Amauri Picollo de; Goetz, Franciane; Nascimento, Vladimir Pinheiro do

    Abstract in English:

    This study assessed biofilm formation on polystyrene by Staphylococcus aureus, Listeria monocytogenes, L. welshimeri and Escherichia coli, isolated from a slaughtering plant, grown on tryptic soy broth (TSB) using different glucose concentrations. The tested bacteria produced biofilm in at least one of the concentrations used, and some of them were strong biofilm producers.
  • In vitro antibacterial activities of ethanol extract of iranian propolis (EEIP) against fish pathogenic bacteria (Aeromonas hydrophila, Yersinia ruckeri & Streptococcus iniae) Veterinary Microbiology

    Tukmechi, Amir; Ownagh, Abdolghaffar; Mohebbat, Ali

    Abstract in English:

    The ''in vitro'' antibacterial activity of ethanol extract of propolis (EEIP) from Urmia, Iran was investigated against three prevalent species of fish bacterial pathogens including: Aeromonas hydrophila LMG 3770, Yersinia ruckeri LMG 3279 and Streptococcus iniae LMG 14520. In this study two standard susceptibility testing techniques (Micro-broth dilution method and Agar-well diffusion method) were used to evaluation of the antibacterial activity of EEIP against the mentioned micro-organisms. Also the chemical composition of propolis was determined by the method of Gas chromatography-mass spectrometry (GC-MS). Twenty-six compounds were identified by gas chromatography-mass spectrometry analysis. Results showed Chemical composition of EEIP contained significant amounts of flavonoids, Sesquiterpenes - mainly Eudesmol and Caryophyllene oxide - aromatic acid, and low amounts of aldehydes and triterpens. Furthermore the ethanol extract of propolis inhibited the growth of all examined micro-organisms with the highest antimicrobial activity against Gram-positive bacteria Streptococcus iniae. Ethanol did not influence the antimicrobial effect of EEIP. These antibacterial properties would warrant further studies on the clinical applications of propolis in aquaculture field.
  • Partial VP2 sequencing of canine parvovirus (CPV) strains circulating in the state of Rio de Janeiro, Brazil: detection of the new variant CPV-2c Veterinary Microbiology

    Castro, T.X.; Costa, E.M; Leite, J.P.G.; Labarthe, N.V.; Cubel Garcia, R.C.N.

    Abstract in English:

    Canine parvovirus (CPV) is the most important enteric virus for dogs and it seems to be undergoing continuous evolution, generating new genetic and antigenic variants throughout the world. The aim of this study was to analyze the distribution of CPV variants from 1995 to 2009 and to investigate the circulation of the new variant CPV-2c in Rio de Janeiro, Brazil. In addition, the clinical features of CPV infection were also reported. After CPV laboratorial confirmation by HA/HI and PCR, thirty-two fecal samples were analyzed by sequencing a 583-bp fragment of the VP2 gene. One sample, collected in 2008 was typed as the new type CPV-2c. All samples from 1995 to 2003 were identified as "new CPV-2a". From 2004 to 2006, both "new CPV-2a" and CPV-2b were observed. From 2006 to 2009, most of the samples were characterized as CPV-2b. The classical signs of CPV enteritis were observed in 16/18 CPV-2a and 5/13 CPV-2b infected puppies. These results show that continuous epidemiological surveillance of CPV strain distribution is essential for studying the patterns of CPV-2a and 2b spread and for determining whether the new variant CPV-2c has become permanently established in Brazilian canine population.
  • Isolation and growth characterization of chlorate and/or bromate resistant mutants generated by spontaneous and induced foreword mutations at several gene loci in Aspergillus niger Microbial Physiology

    Kanan, Ghassan J. M.; Al-Najjar, Heyam E.

    Abstract in English:

    We aimed her mainly to evaluate the contribution of newly employed bromate selection system, in obtaining new Aspergillus niger nitrate/nitrite assimilation defective mutants, through Ultraviolet treatment (UV), 1, 2, 7, 8-Diepoxyoctane (DEO), phenols mixture (Phx)) and spontaneous treatments. The newly employed bromate selection system was able to specify only two putative novel mutant types designated brn (bromate resistant but chlorate sensitive (RS) strain, which may specify nitrite specific transporter) and cbrn mutants (bromate resistant and chlorate resistant strain, which may specify nitrate/nitrite bispecific system). The most relevant and innovative findings of this research work involve the isolation of the RR (cbrn) mutants (a new type of nitrate assimilation defective mutants), that could be useful for studying the bispecific nitrate /nitrite transporter system. The majority of obtained bromate resistant mutants (93.3% of the total mutants obtained by all treatments) were of the brn type, whereas the remaining percentage (6.76%) was given to cbrn strains. The highest percentages of brn mutant strains (48% and 58.6% of the total RS strains) were obtained with UA after spontaneous and Phx treatment, whereas Trp has generated 29% and 42% of RS strains after UV and DEO treatments, respectively. The obtained ratios of cbrn mutants were higher (i.e. in the range of 8.4%-11.64% of the total bromate mutants) with chemical treatments, especially when U.A or Pro was serving as sole N-sources at 25ºC rather than 37ºC. A 69% mutants' yield of Aspergillus niger mutant strains representing nine gene loci (niaD, cnx-6 loci, nrt and nirA) were selected on the bases of chlorate (600 mM) toxicity. All chlorate resistant mutants were completely sensitive to bromate (250 mM). The niaD mutants showed the highest percentage (73.97%) of chlorate resistant mutants obtained with all tested treatments. The UV treatment has generated the highest ratio (86.9%) of niaD mutants, whereas, the least (61%) was obtained with Phx treatment. The highest percentage of cnx mutants (32%) was obtained with Phx treatment. The DEO treatment as compared to other tested treatments was the best to use for obtaining the highest ratios of either nrt (13.8%) mutants or nirA (1.9%) mutants.
  • Some optimal culture conditions for production of cyclosporin a by Fusarium roseum Microbial Physiology

    Ismaiel, Ahmed A.; El-Sayed, El-Sayed A.; Mahmoud, Asmaa A.

    Abstract in English:

    A suitable chemically defined culture medium was selected and some optimal conditions for the production of the highly immunosuppressive compound, cyclosporin A (Cyc A) are reported. Medium of the following composition was favorable for the production of Cyc A by Fusarium roseum: glucose, 30; NaNO3, 2; KH2PO4, 1; MgSO4.7H2O, 0.5 and KCL, 0.5 (g/l). Maximum productivity of Cyc A was achieved at pH 6.0 when 50 ml of the fermentation medium/250 ml flask, inoculated with five fungal agar discs (6 mm, diameter) of 7-days old F. roseum culture after incubation at 30 ºC at 120 rpm for 7 days.
  • Production of extracellular lipases by Rhizopus oligosporus in a stirred fermentor Industrial Microbiology

    Iftikhar, Tehreema; Niaz, Mubashir; Zia, Muhammad Anjum; Haq, Ikram ul

    Abstract in English:

    The present investigation deals with the kinetics of submerged extracellular lipases fermentation by both wild and mutant strains of Rhizopus oligosporus var. microsporus in a laboratory scale stirred fermentor. Other parameters studied were inoculum size, pH, agitation and rate of aeration. It was found that the growth and lipases production was increased gradually and reached its maximum 9.07± 0.42ª U mL-1 (W) and 42.49 ± 3.91ª U mL-1 (M) after 30h of fermentation for both wild and mutant strain. There is overall increase of 109% (W) and 124% (M) in the production of extracellular lipases as compared to shake flask. Another significant finding of the present study is that the fermentation period is reduced to 30 h in case of wild and 23 h in case of mutant from 48 h in shake flask studies. The specific productivity of mutant strain (qp = 377.3 U/g cells/h) was several folds higher than wild strain. The specific production rate and growth coefficient revealed the hyperproducibility of extracellular lipases using mutant IIB-63NTG-7.
  • Improvement of penicillin G acylase expression in Escherichia coli through UV induced mutations Industrial Microbiology

    Arshad, Rubina; Farooq, Shafqat; Ali, Syed Shahid

    Abstract in English:

    We used ultraviolet (UV) radiation to induce mutation in three locally isolated strains of Escherichia coli. Different dilutions of bacterial cultures were exposed to UV lamp of 254 nm wavelength for different time intervals at varied distances ranging from 5 to 210 sec and 5 to 100 cm. Viable colonies were screened for mutants with an increased production of penicillin G acylase (PGA) and a reduced production of β-lactamase, which are the desired properties of PGA producing industrial strains. A survival curve was made to get optimum exposure time and distance. The survival percentage for each exposure period was calculated and 1-5% survival was found useful for obtaining mutants with desired change. Screening for PGA and β-lactamase constitutive and/or deficient mutants was made by Serratia marcescens overlay test. A total of 100 survivors were selected of which 49% expressed PGA activity higher than the parent strain. Frequency of β-lactamase constitutive and deficient mutants was 48 and 52%, respectively. The best hyper-producing mutant (BDCS-N-M74), with almost negligible expression of β-lactamase, exhibited three-fold (22.5 mg 6-APA h-1 mg-1 wet cells) increase in PGA activity compared with that in the parent strain (6.7 mg 6-APA h-1 mg-1 wet cells). The results indicated the successful induction of UV mediated mutation in E. coli for PGA hyper-producing mutants lacking β-lactamase activity.
  • Application of representational difference analysis to identify genomic differences between Bradyrhizobium elkanii and B. Japonicum species Genetics And Molecular Microbiology

    Soares, René Arderius; Passaglia, Luciane Maria Pereira

    Abstract in English:

    Bradyrhizobium elkanii is successfully used in the formulation of commercial inoculants and, together with B. japonicum, it fully supplies the plant nitrogen demands. Despite the similarity between B. japonicum and B. elkanii species, several works demonstrated genetic and physiological differences between them. In this work Representational Difference Analysis (RDA) was used for genomic comparison between B. elkanii SEMIA 587, a crop inoculant strain, and B. japonicum USDA 110, a reference strain. Two hundred sequences were obtained. From these, 46 sequences belonged exclusively to the genome of B. elkanii strain, and 154 showed similarity to sequences from B. japonicum genome. From the 46 sequences with no similarity to sequences from B. japonicum, 39 showed no similarity to sequences in public databases and seven showed similarity to sequences of genes coding for known proteins. These seven sequences were divided in three groups: similar to sequences from other Bradyrhizobium strains, similar to sequences from other nitrogen-fixing bacteria, and similar to sequences from non nitrogen-fixing bacteria. These new sequences could be used as DNA markers in order to investigate the rates of genetic material gain and loss in natural Bradyrhizobium strains.
Sociedade Brasileira de Microbiologia USP - ICB III - Dep. de Microbiologia, Sociedade Brasileira de Microbiologia, Av. Prof. Lineu Prestes, 2415, Cidade Universitária, 05508-900 São Paulo, SP - Brasil, Ramal USP 7979, Tel. / Fax: (55 11) 3813-9647 ou 3037-7095 - São Paulo - SP - Brazil
E-mail: bjm@sbmicrobiologia.org.br