Genotyping of the MTL loci and susceptibility to two antifungal agents of Candida glabrata clinical isolates

The opportunistic fungal pathogen Candida glabrata is the second most common isolate from bloodstream infections worldwide and is naturally less susceptible to the antifungal drug fluconazole than other Candida species. C. glabrata is a haploid yeast that contains three mating-type like loci (MTL), although no sexual cycle has been described. Strains containing both types of mating information at the MTL1 locus are found in clinical isolates, but it is thought that strains containing type a information are more common. Here we investigated if a particular combination of mating type information at each MTLlocus is more prevalent in clinical isolates from hospitalized patients in Mexico and if there is a correlation between mating information and resistance to fluconazole and 5-fluorocytosine. We found that while both types of information at MTL1 are equally represented in a collection of 64 clinical isolates, the vast majority of isolates contain a-type information at MTL2 and α-type at MTL3. We also found no correlation of the particular combination of mating type information at the three MTL loci and resistance to fluconazole.

Susceptibility of Mycobacterium tuberculosis to first-line antimycobacterial agents in a Brazilian hospital: assessing the utility of the tetrazolium (MTT) microplate assay

We conducted a cross-sectional, hospital-based study between January 2006-March 2008 to estimate the resistance of Mycobacterium tuberculosis to first-line drugs in patients with tuberculosis at a Brazilian hospital. We evaluated the performance of the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] (MTT) microplate assay compared with the Bactec-MGIT 960™ system for mycobacteria testing. The prevalence of resistance in M. tuberculosis was 6.7%. Multidrug-resistance [resistance to rifampicin (RMP) and isoniazid (INH)], INH-resistance and streptomycin (SM)-resistance accounted for 1%, 3.8% and 3.8% of all resistance, respectively, and all isolates were susceptible to ethambutol (EM). The resistance was primary in four cases and acquired in three cases and previous treatment was associated with resistance (p = 0.0129). Among the 119 M. tuberculosis isolates, complete concordance of the results for INH and EM was observed between the MTT microplate and Bactec-MGIT 960TM methods. The observed agreement for RMP was 99% (sensitivity: 90%) and 95.8% for SM (sensitivity 90.9%), lower than those for other drugs. The MTT colourimetric method is an accurate, simple and low-cost alternative in settings with limited resources.

Evaluation of the synergistic potential of vancomycin combined with other antimicrobial agents against methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus spp strains

Methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative Staphylococcus spp (CNS) are the most common pathogens that cause serious long term infections in patients. Despite the existence of new antimicrobial agents, such as linezolid, vancomycin (VAN) remains the standard therapy for the treatment of infections caused by these multidrug-resistant strains. However, the use of VAN has been associated with a high frequency of therapeutic failures in some clinical scenarios, mainly with decreasing concentration of VAN. This work aims to evaluate the synergic potential of VAN plus sulfamethoxazole/trimethoprim (SXT), VAN plus rifampin (RIF) and VAN plus imipenem (IPM) in sub-minimum inhibitory concentrations against 22 clinical strains of MRSA and CNS. The checkerboard method showed synergism of VAN/RIF and VAN/SXT against two and three of the 22 strains, respectively. The combination of VAN with IPM showed synergistic effects against 21 out of 22 strains by the E-test method. Four strains were analyzed by the time-kill curve method and synergistic activity was observed with VAN/SXT, VAN/RIF and especially VAN/IPM in sub-inhibitory concentrations. It would be interesting to determine if synergy occurs in vivo. Evidence of in vivo synergy could lead to a reduction of the standard VAN dosage or treatment time.

Genotypes of Cryptococcus neoformans and Cryptococcus gattii as agents of endemic cryptococcosis in Teresina, Piauí (northeastern Brazil)

Throughout Brazil, Cryptococcus neoformans is the cause of cryptococcosis, whereas Cryptococcus gattii is endemic to the northern and northeastern states. In this study, the molecular types of 63 cryptococcal isolates recovered from the cerebrospinal fluid of meningitis patients diagnosed between 2008-2010 in Teresina, Piauí, Brazil, were analysed. Out of the 63 patients, 37 (58.7%) were human immunodeficiency virus (HIV)-positive and 26 (41.3%) were HIV-negative. URA5-restriction fragment length polymorphism analysis identified 37/63 (58.7%) isolates as the C. neoformans VNI genotype, predominantly in HIV-positive patients (32/37, 86.5%), and 24/63 (38.1%) as the C. gattii VGII genotype, mostly in HIV-negative patients (21/26, 80.8%). The occurrence of C. gattii VGII in six apparently healthy children and in seven adolescents/young adults in this region reaffirms the endemic occurrence of C. gattii VGII-induced primary cryptococcosis and early cryptococcal infection. Lethality occurred in 18/37 (48.6%) of the HIV-positive subjects and in 13/26 (50%) of the HIV-negative patients. Our results provide new information on the molecular epidemiology of C. neoformans and C. gattii in Brazilian endemic areas.

Effects of the encapsulation of usnic acid into liposomes and interactions with antituberculous agents against multidrug-resistant tuberculosis clinical isolates

Mycobacterium tuberculosis (Mtb) has acquired resistance and consequently the antibiotic therapeutic options available against this microorganism are limited. In this scenario, the use of usnic acid (UA), a natural compound, encapsulated into liposomes is proposed as a new approach in multidrug-resistant tuberculosis (MDR-TB) therapy. Thus the aim of this study was to evaluate the effect of the encapsulation of UA into liposomes, as well as its combination with antituberculous agents such as rifampicin (RIF) and isoniazid (INH) against MDR-TB clinical isolates. The in vitro antimycobacterial activity of UA-loaded liposomes (UA-Lipo) against MDR-TB was assessed by the microdilution method. The in vitro interaction of UA with antituberculous agents was carried out using checkerboard method. Minimal inhibitory concentration values were 31.25 and 0.98 µg/mL for UA and UA-Lipo, respectively. The results exhibited a synergistic interaction between RIF and UA [fractional inhibitory concentration index (FICI) = 0.31] or UA-Lipo (FICI = 0.28). Regarding INH, the combination of UA or UA-Lipo revealed no marked effect (FICI = 1.30-2.50). The UA-Lipo may be used as a dosage form to improve the antimycobacterial activity of RIF, a first-line drug for the treatment of infections caused by Mtb.

Functional complementation of Leishmania (Leishmania) amazonensis AP endonuclease gene (lamap) in Escherichia coli mutant strains challenged with DNA damage agents

During its life cycle Leishmania spp. face several stress conditions that can cause DNA damages. Base Excision Repair plays an important role in DNA maintenance and it is one of the most conserved mechanisms in all living organisms. DNA repair in trypanosomatids has been reported only for Old World Leishmania species. Here the AP endonuclease from Leishmania (L.) amazonensis was cloned, expressed in Escherichia coli mutants defective on the DNA repair machinery, that were submitted to different stress conditions, showing ability to survive in comparison to the triple null mutant parental strain BW535. Phylogenetic and multiple sequence analyses also confirmed that LAMAP belongs to the AP endonuclease class of proteins.

Supervision of Community Health Agents in the Family Health Strategy: the perspective of nurses

Objective Analyzing the conceptions of supervision of nurses in the Family Health Strategy (ESF - Estratégia Saúde da Família) in relation to community health workers (ACS – Agente Comunitário de Saúde), taking for reference the work process and the power relations. Method A qualitative study, in which 18 interviews were carried out with nurses from the ESF in the countryside of the state of Goiás. Following transcription, the data were subjected to thematic content analysis and the following categories were identified: Supervision with that sees it all and Supervision is participating and working together. Results Among the findings, it is clear the character of control and monitoring that is carried out primarily through printed instruments and home visits, which constitute vertical and impositive relations, and suffer influence of the local health management. Another finding is the supervision as acting together and with the educational perspective. Conclusion The supervision carried out does not provide the changes intended by the ESF.

Social representation of domestic violence against women among Nursing Technicians and Community Agents

OBJECTIVE To analyze the social representations of the Nursing Technicians and Community Health Agents about domestic violence against women. METHOD A qualitative study carried out in the city of Rio Grande, RS, in which evocations and interviews were collected between July and November 2013. For the treatment of data were used the EVOC 2005 software and the context analysis. RESULT It is a structured representation, in which the central nucleus contains conceptual, imaging and attitudinal elements, namely: abuse, aggression, physical aggression, cowardice and lack of respect. Such terms were present in the context of the interviews. The professionals acknowledged that violence is not limited to physical aspects and were judgemental about the acts of the aggressor. CONCLUSION This knowledge may enable the problematization of the studied phenomenon with the team, and facilitate the search for prevention and intervention strategies for victims, offenders and managers of health services.

Workloads and strain process in Community Health Agents

ABSTRACT OBJECTIVE To identify the workloads present in the work activities of community health agents (CHAs) and the resulting strain processes. METHOD A descriptive, exploratory, cross-sectional and quantitative study conducted with 137 CHAs. Data were collected through a questionnaire and interview guided by the health surveillance software called SIMOSTE (Health Monitoring System of Nursing Workers), following the ethical codes of the current law. RESULTS In total, were identified 140 workloads involved in 122 strain processes, represented by the occurrence of health problems of the CHAs. The mechanical (55.00%) and biological (16.43%) loads stood out. The most common strain processes were the external causes of morbidity and mortality (62.31%) and diseases of the musculoskeletal system and connective tissue (10.66%). CONCLUSION From the identified overloads, it became evident that all workloads are present in the work process of CHAs, highlighting the mechanical load, represented mainly by external causes of morbidity and mortality that are related to occupational accidents.

A 15-year post evaluation of the fire effects on ant community in an area of Amazonian forest

Fire represents an important disturbance to ant communities in areas of fire regime. Otherwise, little is known about the effects of fire on ant communities in areas of non-fire regimes, such as in the Amazonian region. We evaluated the long-term effect of fire on ant species richness in a rain forest (Bacaba Plateau) burned 15-years ago and compare our data with the data of primary unburned forest. A total of 85 ant species distributed in 21 genera and 14 tribes were collected; among them, 72 and 44 species were found on the litter and vegetation, respectively. The fire damaged forest studied supports an intermediate richness of ants when compared to a primary unburned rain forest in the same region. A comparative analysis of ant species richness showed that the Bacaba Plateau presented a different ant fauna when compared with the primary unburned forests, suggesting that fire can alter ant species composition. Although, our results cannot be conclusive on the effects of fire on ant community, they represent a pioneer data on human induced fire in tropical rain forests.

Fusarium oxysporum strains as biocontrol agents against Fusarium wilt: effects on soil microbial biomass and activity

Before planning the large-scale use of nonpathogenic strains of Fusarium oxysporum as biocontrol agents of Fusarium wilt, their behaviour and potential impact on soil ecosystems should be carefully studied as part of risk assessment. The aim of this work was to evaluate the effects of antagonistic F. oxysporum strains, genetically manipulated (T26/6) or not (233/1), on soil microbial biomass and activity. The effects were evaluated, in North-western Italy, in two soils from different sites at Albenga, one natural and the other previously solarized, and in a third soil obtained from a 10-year-old poplar stand (Popolus sp.), near Carignano. There were no detectable effects on ATP, fluorescein diacetate hydrolysis, and biomass P that could be attributed to the introduction of the antagonists. A transient increase of carbon dioxide evolution and biomass C was observed in response to the added inoculum. Although the results showed only some transient alterations, further studies are required to evaluate effects on specific microorganism populations.

Effects of fire on soil nitrogen dynamics and microbial biomass in savannas of Central Brazil

The objective of this work was to study the effects of fire on net N mineralization and soil microbial biomass in burned and unburned cerrado stricto sensu sites. The study was carried out from April 1998 to April 2000. The pH values were significantly higher in the burned site while soil moisture content was significantly higher in the unburned site (P<0.05). The soil C/N ratio was 22/1 and the available NO3-N ranged between 1.5 and 2.8 mg kg-¹ dry weight. However, the NH4-N concentration ranged between 3 and 34 mg kg-1 dry weight in the burned site and between 3 and 22 mg kg-1 dry weight in the unburned site. The NH4-N increased after fire, but no significant changes were observed for NO3-N (P<0.05). The NO3-N accumulation occurred in short periods during the rainy season. The rates of net N mineralization increased during the rainy season while reductions in soil microbial biomass were observed at both sites. This suggested that the peak in microbial activities occurred with the first rain events, with an initial net immobilization followed by net mineralization. Both sites presented the same pattern for mineralization/immobilization, however, the amount of inorganic-N cycled annually in unburned site was 14.7 kg ha-1 per year while the burned site presented only 3.8 kg ha-¹ of inorganic-N, one year after the burning.

Microbial communities in Cerrado soils under native vegetation subjected to prescribed fire and under pasture

The objective of this work was to evaluate the effects of fire regimes and vegetation cover on the structure and dynamics of soil microbial communities, through phospholipid fatty acid (PLFA) analysis. Comparisons were made between native areas with different woody covers ("cerrado stricto sensu" and "campo sujo"), under different fire regimes, and a 20-year-old active palisadegrass pasture in the Central Plateau of Brazil. Microbial biomass was higher in the native plots than in the pasture, and the highest monthly values were observed during the rainy season in the native plots. No significant differences were observed between fire regimes or between communities from the two native vegetation types. However, the principal component (PC) analysis separated the microbial communities by vegetation cover (native x pasture) and season (wet x dry), accounting for 45.8% (PC1 and PC3) and 25.6% (PC2 and PC3), respectively, of the total PLFA variability. Changes in land cover and seasonal rainfall in Cerrado ecosystems have significant effects on the total density of soil microorganisms and on the abundance of microbial groups, especially Gram-negative and Gram-positive bacteria.

Soil organic matter in fire-affected pastures and in an Araucaria forest in South-Brazilian Leptosols

The objective of this work was to evaluate the distribution pattern and composition of soil organic matter (SOM) and its physical pools of Leptosols periodically affected by fire over the last 100 years in South Brazil. Soil samples at 0-5, 5-10, and 10-15 cm depths were collected from the following environments: native pasture without burning in the last year and grazed with 0.5 livestock per hectare per year (1NB); native pasture without burning in the last 23 years and grazed with 2.0 livestock per hectare per year (23NB); and an Araucaria forest (AF). Physical fractionation was performed with the 0-5 and 5-10 cm soil layers. Soil C and N stocks were determined in the three depths and in the physical pools, and organic matter was characterized by infrared spectroscopy and thermogravimetry. The largest C stocks in all depths and physical pools were found under the AF. The 23NB environment showed the lowest soil C and N stocks at the 5-15 cm depth, which was related to the end of burning and to the higher grazing intensity. The SOM of the occluded light fraction showed a greater chemical recalcitrance in 1NB than in 23NB. Annual pasture burning does not affect soil C stocks up to 15 cm of depth.