A comparison of mycolic acid analysis for nontuberculous mycobacteria identification by thin-layer chromatography and molecular methods

The development of fast, inexpensive, and reliable tests to identify nontuberculous mycobacteria (NTM) is needed. Studies have indicated that the conventional identification procedures, including biochemical assays, are imprecise. This study evaluated a proposed alternative identification method in which 83 NTM isolates, previously identified by conventional biochemical testing and in-house M. avium IS1245-PCR amplification, were submitted to the following tests: thin-layer chromatography (TLC) of mycolic acids and PCR-restriction enzyme analysis of hsp65 (PRA). High-performance liquid chromatography (HPLC) analysis of mycolic acids and Southern blot analysis for M. avium IS1245 were performed on the strains that evidenced discrepancies on either of the above tests. Sixty-eight out of 83 (82%) isolates were concordantly identified by the presence of IS1245 and PRA and by TLC mycolic acid analysis. Discrepant results were found between the phenotypic and molecular tests in 12/83 (14.4%) isolates. Most of these strains were isolated from non-sterile body sites and were most probably colonizing in the host tissue. While TLC patterns suggested the presence of polymycobacterial infection in 3/83 (3.6%) cultures, this was the case in only one HPLC-tested culture and in none of those tested by PRA. The results of this study indicated that, as a phenotypic identification procedure, TLC mycolic acid determination could be considered a relatively simple and cost-effective method for routine screening of NTM isolates in mycobacteriology laboratory practice with a potential for use in developing countries. Further positive evidence was that this method demonstrated general agreement on MAC and M. simiae identification, including in the mixed cultures that predominated in the isolates of the disseminated infections in the AIDS patients under study. In view of the fact that the same treatment regimen is recommended for infections caused by these two species, TLC mycolic acid analysis may be a useful identification tool wherever molecular methods are unaffordable.

The temporal resolution of epibiont assemblages: Are they ecological snapshots or overexposures?

The effects of time averaging on the fossil record of soft-substrate marine faunas have been investigated in great detail, but the temporal resolution of epibiont assemblages has been inferred only from limited-duration deployment experiments. Individually dated shells provide insight into the temporal resolution of epibiont assemblages and the taphonomic history of their hosts over decades to centuries. Epibiont abundance and richness were evaluated for 86 dated valves of the rhynchonelliform brachiopod Bouchardia rosea collected from the inner shelf. Maximum abundance occurred on shells less than 400 yr old, and maximum diversity was attained within a century. Taphonomic evidence does not support models of live-host colonization, net accumulation, or erasure of epibionts over time. Encrustation appears to have occurred during a brief interval between host death and burial, with no evidence of significant recolonization of exhumed shells. Epibiont assemblages of individually dated shells preserve ecological snapshots, despite host-shell time averaging, and may record long-term ecological changes or anthropogenic environmental changes. Unless the ages of individual shells are directly estimated, however, pooling shells of different ages artificially reduces the temporal resolution of their encrusting assemblages to that of their hosts, an artifact of analytical time averaging. © 2006 by The University of Chicago. All rights reserved.

Efeito da adição de probiótico e/ou prebiótico em dietas de leitões desmamados sobre o desempenho, incidência de diarréia e contagem de coliformes totais

In the Experiment 1, 50 pigs weaned at the 21 days were fed with either of five diets: T1 - basal diet; T2 - basal diet + antibiotic; T3 - basal diet + probiotic; T4 - basal diet + prebiotic; T5 - basal diet + simbiotic. The variables studied were body weight, feed intake and feed conversion in the Phase 1 (21 to 43 days), Phase 2 (44 to 57 days), Phase 3 (58 to 70 days) and Total Phase (21 to 70 days). During the performance experiment, a fecal survey score was conducted to verify diarrhea incidence. In the Experiment 2, 44 pigs weaned at 21 days were fed with the same diets of Experiment 1. Pigs were slaughtered at three differents ages (at weaning, at seven and 14 days after weaning). A segment of the small intestine was collected for analisys of total coliformes. The results showed that the best performance was obtained with the utilization of prebiotic and simbiotic. There were not differences in relation to diarrhea incidence among the treatments studied. The addition of probiotic and/or prebiotic in the diet prevented increase of colonization by pathogenic bacterias from seven to 14 days after weaning.

Presence of Candida spp. in infants oral cavity and its association with early childhood caries

Candida spp., mainly C. albicans, colonizes oral cavity of infants. Transmission by mother to childbirth, pacifier use, feeding habits and caries are factors related to Candida oral colonization. Some researches related that early childhood caries favor the oral colonization of C. albicans. The present literature review described the presence of Candida spp. in oral cavity of infants and its association with early childhood caries (ECC). The literature was searched for original papers relating Candida, pacifier and baby bottle usage and ECC.The articles, were selected using Bireme and Medline databases. Manual tracing of references cited in key papers was also elicited It can be concluded that Candida spp. colonization in the infants'oral cavity especially C. albicans, can be related to the pacifier usage, feeding habits caries lesion. The early childhood caries favor the C. albicans colonization, although it's role in the carious process need further studies to be elucidate scientifically.

Comparative study of the clinical and anti-microbial efficacy of tongue cleaners

Candida species have frequently been isolated from the oral cavities of a variety of patients, such as elderly people, dentures users, immunocompromised and health patients. Yeasts may be associated with immune response and local factors such as poor oral hygiene. It was evaluated effectiveness of tongue cleaner showing which types would be preferred by patients, changes in tongue coating and in saliva yeasts counting. Thirty patients were selected and randomly distributed into three groups. This crossover blind study evaluated the effect of tongue cleaning using: a plastic and a steel tongue scraper and a nylon soft-bristle toothbrush. All patients were instructed to use the cleaners twice a day for one week (fifteen-day wash-out period). Saliva and tongue coating samples were collected from each patient from each test period, the yeasts were counted by colony forming units per mL (CFU/ mL) and the species were identified. The patients were questioned about cleaner preference. An increase in the percentage of patients with no tongue coating after scraping was observed. A reduction in the mean number of Candida species in tongue coating was observed only after nylon soft-bristle toothbrush cleaner. Candida albicans was the prevalent species. Volunteers preferred to the steel tongue scraper (60%). Tongue cleaners reduced the tongue coating and the mean number of saliva's yeasts. Degree of tongue coating favors the Candida species colonization.

Effect of an alcoholic diet on dental caries and on Streptococcus of the mutans group: Study in rats

The objective of this study was to evaluate the effects of an alcohol diet on Streptococcus of the mutans group and on dental caries in the oral cavity of rats. Forty animals were divided into 3 groups according to the following liquid diets: 20% ethanol solution (Alcohol Group, AG), 27% sucrose solution (Isocaloric Group, IG), and water (Control Group, CG). After 56 days, samples were collected and plated on Mitis Salivarius Bacitracin agar to assess the number of colony forming units (CFU/mL) of Streptococcus of the mutans group. The animals were sacrificed and the jaws were removed in order to assess the occurrence of dental caries on the smooth and occlusal surfaces using stereomicroscopy. The data were submitted to ANOVA and Tukey test. The average numbers of CFU/mL (10 3) were: 8.17 (AG), 9.78 (IG), and 5.63 (CG). There was no significant difference among the groups for the occurrence of occlusal caries. Regarding smooth surface caries, in the upper jaw, the caries number in the IG (1.58) was similar to that in the AG (2.06) and in the CG (1.14), and the number of caries in the AG was higher than in the CG; in the lower jaw there was significant difference among the 3 groups: AG (1.14), IG (2.00) and CG (0.43). The diets with the alcohol and sucrose solutions presented a tendency of increasing the colonization by Streptococcus of the mutans group and of increasing the occurrence of smooth surface dental caries in rat molars when compared to the control diet.

Morfogênese do testículo de embriões e fetos de vacas da raça nelore (Bos taurus indicus)

The aim of this study was to accompany the process of testicular development from the non-differentiable phase to its complete formation. Embryos and fetuses of Nelore breed cows (Bos Taurus indicus) were obtained in slaughterhouses near the Uberlandia city, Minas Gerais. The gonads and the embryos were fixed in Bouin's fixative and afterwards processed for conventional optical microscopy. The gonadal was observed firstly in a 1.0 cm long embryo. In 2.5 cm long embryos the presence of the albuginea allows the sex identification. The mean thickness of the albuginea ranged from 29.08 to 558.45 mm. Gradually increase of vascularization of the albuginea and parenchyma is observed. The mediastinum is located centrally. There was a decrease in the space occupied by the testicular cords, from 63.71 to 41.99% of the total testes volume. Its diameter ranged from 31.68 to 48.80 mm. The diameter of germinal cells (and their nuclei) was from 12.27 (6.65) to 16.95 914.21) mm. The quantity of germinal cells by cross section of cord decreased from a maximum of 2.80 to 0.76. The total number of germinal cells was from 16 at the beginning of colonization of the gonad to 18.32 x 106 at the end of the study. The number of Sertoli's cells by cross section of cord ranged from 10.00 to 16.25. The results obtained show that the origin and formation of testes in embryos and fetuses from Nelore breed cows (Bos taurus indicus) does occur in a very similar way to what is described for Bos taurus taurus.

Biotic interaction between spionid polychaetes and bouchardiid brachiopods: Paleoecological, taphonomic and evolutionary implications

Shells of Bouchardia rosea (Brachiopoda, Rhynchonelliformea) are abundant in Late Holocene death assemblages of the Ubatuba Bight, Brazil, SW Atlantic. This genus is also known from multiple localities in the Cenozoic fossil record of South America. A total of 1211 valves of B. rosea, 2086 shells of sympatric bivalve mollusks (14 nearshore localities ranging in depth from 0 to 30 m), 80 shells of Bouchardia zitteli, San Julián Formation, Paleogene, Argentina, and 135 shells of Bouchardia transplatina, Camacho Formation, Neogene, Uruguay were examined for bioerosion traces. All examined bouchardiid shells represent shallow-water, subtropical marine settings. Out of 1211 brachiopod shells of B. rosea, 1201 represent dead individuals. A total of 149 dead specimens displayed polychaete traces (Caulostrepsis). Live polychaetes were found inside Caulostrepsis borings in 10 life-collected brachiopods, indicating a syn-vivo interaction (Caulostrepsis traces in dead shells of B. rosea were always empty). The long and coiled peristomial palps, large chaetae on both sides of the 5th segment, and flanged pygidium found in the polychaetes are characteristic of the polychaete genus Polydora (Spionidae). The fact that 100% of the Caulostrepsis found in living brachiopods were still inhabited by the trace-making spionids, whereas none was found in dead hosts, implies active biotic interaction between the two living organisms rather than colonization of dead brachiopod shells. The absence of blisters, the lack of valve/site stereotypy, and the fact that tubes open only externally are all suggestive of a commensal relationship. These data document a new host group (bouchardiid rhynchonelliform brachiopods) with which spionids can interact (interestingly, spionid-infested sympatric bivalves have not been found in the study area despite extensive sampling). The syn-vivo interaction indicates that substantial bioerosion may occur when the host is alive. Thus, the presence of such bioerosion traces on fossil shells need not imply a prolonged post-mortem exposure of shells on the sea floor. Also, none of the Paleogene and Neogene Bouchardia species included any ichnological evidence for spionid infestation. This indicates that the Spionidae/ Bouchardia association may be geologically young, although the lack of older records may also reflect limited sampling and/or taphonomic biases.

Modelos dinâmicos de assembléia

The structure of an ecological community is shaped by several temporally varying mechanisms. Such mechanisms depend in a large extent on species interactions, which are themselves manifestations of the community's own structure. Dynamics and structure are then mutually determined. The assembly models are mathematical or computational models which simulate the dynamics of ecological communities resulting from a historical balance among colonizations and local extinctions, by means of sequential species introductions and their interactions with resident species. They allow analyzing that double relationship between structure and dynamics, recognizing its temporal dependence. It is assumed two spatiotemporal scales: (i) a local scale, where species co-occur and have their dynamics explicitly simulated and (ii) a regional scale without dynamics, representing the external environment which the potential colonizers come from. The mathematical and computational models used to simulate the local dynamics are quite variable, being distinguished according to the complexity mode of population representation, including or not intra or interspecific differences. They determine the community state, in terms of abundances, interactions, and extinctions between two successive colonization attempts. The schedules of species introductions also follow diverse (although arbitrary) rules, which vary qualitatively with respect to species appearance mode, whether by speciation or by immigration, and quantitatively with respect to their rates of introduction into the community. Combining these criteria arises a great range of approaches for assembly models, each with its own limitations and questions, but contributing in a complementary way to elucidate the mechanisms structuring natural communities. To present such approaches, still incipient as research fields in Brazil, to describe some methods of analysis and to discuss the implications of their assumptions for the understanding of ecological patterns are the objectives of the present review.

Pasteuriza̧ão de turfa brasileira para o cultivo de Agaricus brasiliensis

Agaricus brasiliensis is a Brazilian basidiomycete which has been cultivated and consumed around the world as a therapeutic food. Casing layer is one of the most important steps on A. brasiliensis cultivation and European peat is the most used casing layer on Agaricus bisporus cultivation. Besides the importance of it on mushroom cultivation the peat import could be a risk of exotic organism introduction. Alternative as Brazilian peat is barely used for mushroom growers in Brazil. Thus, the objective of this work was to evaluate Brazilian peat with and without pasteurization as casing layer on A. brasiliensis cultivation. The fungus was previously grown on wheat grains and transferred to a substratum prepared by composted traditional method. After mycelium colonization of the substratum a pasteurized or non pasteurized Brazilian peat (casing layer) was added. It was concluded that pasteurization of the casing layer increases in 30% yield after 65 days of cultivation. There is no difference of yield for pasteurized and non pasteurized casing layer until 30 days of cultivation. An increase of flies is observed in non pasteurized casing layer. The production flush is easily perceived with pasteurized casing layer but not with non pasteurized casing layer.

Salmonella enterica serovar typhimurium colonizing the lumen of the chicken intestine grows slowly and upregulates a unique set of virulence and metabolism genes

The pattern of global gene expression in Salmonella enterica serovar Typhimurium bacteria harvested from the chicken intestinal lumen (cecum) was compared with that of a late-log-phase LB broth culture using a whole-genome microarray. Levels of transcription, translation, and cell division in vivo were lower than those in vitro. S. Typhimurium appeared to be using carbon sources, such as propionate, 1,2-propanediol, and ethanolamine, in addition to melibiose and ascorbate, the latter possibly transformed to D-xylulose. Amino acid starvation appeared to be a factor during colonization. Bacteria in the lumen were non- or weakly motile and nonchemotactic but showed upregulation of a number of fimbrial and Salmonella pathogenicity island 3 (SPI-3) and 5 genes, suggesting a close physical association with the host during colonization. S. Typhimurium bacteria harvested from the cecal mucosa showed an expression profile similar to that of bacteria from the intestinal lumen, except that levels of transcription, translation, and cell division were higher and glucose may also have been used as a carbon source. © 2011, American Society for Microbiology.

A moderna agricultura no final do século XIX em São Paulo: algumas propostas

Esta pesquisa se insere dentro da história da agricultura no Brasil, especialmente no Estado de São Paulo, a partir da segunda metade do século XIX, início do século XX. As propostas de modernização agrícola giravam em torno de diferentes assuntos, mas sempre interligados, como: diversificação da agricultura, adubação química e natural, cultura intensiva do solo, mão de obra, povoamento e colonização, instalação de núcleos coloniais dentro dos moldes da moderna agricultura então desejada, instrução agrícola por meio de campos de experiências e demonstração, ensino agrícola para diferentes graus, a mecanização da lavoura, o que era chamado, na época, de Moderna Agricultura, em substituição ao que era considerado como agricultura rotineira ou atrasada, praticada desde o início da agricultura no Brasil.

Effect of brushing with conventional versus whitening dentifrices on surface roughness and biofilm formation of dental ceramics

The aim of this study was to evaluate the effect of conventional and whitening dentifrices on the weight loss, surface roughness, and early in situ biofilm formation on the surface of dental ceramics. Standardized feldspar ceramic specimens (Vita VM7 and Vita VM13) were submitted to the following experimental conditions: no brushing; brushing without a dentifrice; brushing with a conventional dentifrice; and brushing with a whitening dentifrice. A brushing machine was used to simulate brushing. The mass and surface roughness of all specimens from the test groups were evaluated prior to and after brushing. Ten participants used an oral device for eight hours to evaluate the biofilm formed in situ on the specimens. Scanning electron microscopy was used for qualitative and quantitative analysis of the biofilm. ANOVA and Tukey tests were used to analyze the results of weight loss, surface roughness, and presence of bacteria. A one-way Kruskal-Wallis test was used for bacterial colonization results. For both ceramics, brushing with a whitening dentifrice resulted in weight loss that was significantly greater when compared to brushing without a dentifrice or with a conventional dentifrice. Increased surface roughness was noticed on VM13 ceramic samples with both dentifrices, whereas only conventional dentifrice had a significant effect on the surface roughness of VM7 samples. For both VM7 and VM13, no difference was found between the experimental conditions with regard to the presence or number of bacteria. Cocci and short rods were the predominant microbial morphotypes. Granular or fibrillar acellular material partially covered the specimens. Brushing with a whitening dentifrice resulted in significant weight loss of ceramic restorations, while brushing with both conventional and whitening dentifrices can roughen ceramic surfaces. The increase in roughness was not clinically significant to contribute to increased biofilm formation.

Modernization and new services of the Brazilian active control network

The Brazilian Network for Continuous Monitoring of GNSS - RBMC is a national network of continuously operating reference GNSS stations. Since its establishment in December of 1996, it has been playing an essential role for the maintenance and user access of the fundamental geodetic frame in the country. In order to provide better services for RBMC, the Brazilian Institute of Geography and Statistics - IBGE and the National Institute of Colonization and Land Reform - INCRA are both partners involved in the National Geospatial Framework Project - PIGN. This paper provides an overview of the recent modernization phases the RBMC network has undergone highlighting its future steps. These steps involve the installation of new equipment, provide real time data from a group of core stations and compute real-time DGPS corrections, based on CDGPS (The real-time Canada-Wide DGPS Service) (The Real-Time Canada-Wide DGPS Service. http://www.cdgps.com/ 2009a). In addition to this, a post-mission Precise Point Positioning (PPP) service has been established based on the current Geodetic Survey Division of NRCan (CSRS-PPP) service. This service is operational since April 2009 and is in large use in the country. All activities mentioned before are based on a cooperation signed at the end of 2004 with the University of New Brunswick, supported by the Canadian International Development Agency and the Brazilian Cooperation Agency. The Geodetic Survey Division of NRCan is also participating in this modernization effort under the same project. This infrastructure of 66 GNSS stations, the real time, post processing services and the potentiality of providing Wide Area DGPS corrections in the future show that the RBMC system is comparable to those available in USA and Europe. © Springer-Verlag Berlin Heidelberg 2012.

Avaliação microbiológica da superfície dos implantes ortopédicos metálicos usados para fixar fraturas ósseas

In the majority of cases of bone fracture requiring surgery, orthopedic implants (screw-plate and screw) are used for osteosynthesis and the infections associated with such implants are due to the growth of microorganisms in biofilms. The objective of this study was to identify microorganisms recovered from osteosynthesis implants used to fix bone fractures, to assess the viability of the cells and the ability of staphylococci to adhere to a substrate and to determine their sensitivity/resistance to antimicrobials. After surgical removal, the metal parts of austenitic stainless steel (ASTM F138/F139 or ISO NBR 5832-1/9) were transported to the Laboratory of Clinical Microbiology, washed in buffer and subjected to ultrasonic bath at 40±2 kHz for 5 minutes. The sonicated fluid was used to seed solid culture media and cell viability was assessed under the microscope by with the aid of a fluorescent marker. The production of extracellular polysaccharide by Staphylococcus spp. was investigated by means of adhesion to a polystyrene plate. The profile of susceptibility to antimicrobials was determined by the disk diffusion assay. The most frequently isolated bacteria included coagulase-negative Staphylococcus resistant to erythromycin, clindamycin and oxacillin. Less frequent were Pseudomonas aeruginosa resistant to trimethoprim/sulfamethoxazole and ampicillin, Acinetobacter baumannii resistant to ceftazidime, Enterobacter cloacae resistant to cephalothin, cefoxitin, cefazolin, levofloxacin and ciprofloxacin, Bacillus spp. and Candida tropicalis. The observation of slides by fluorescence microscope showed clusters of living cells embedded in a transparent matrix. The test for adherence of coagulase-negative Staphylococcus to a polystyrene plate showed that these microorganisms produce extracellular polysaccharide. In conclusion, the metal parts were colonized by bacteria related to orthopedic implant infection, which were resistant to multiple antibiotics.