Phytochemical,Antiplasmodial, Cytotoxic and Antimicrobial Evaluation of a Southeast Brazilian Brown Propolis Produced by Apis mellifera Bees

Seven phenolic compounds (ferulic acid, caffeic acid, 4-methoxycinnamic acid, 3,4-dimethoxycinnamic acid, 3-hydroxy-4-methoxybenzaldehyde, 3-methoxy-4-hydroxypropiophenone and 1-O,2-O-digalloyl-6-O-trans-p-coumaroyl-β-D-glucopyranoside), a flavanonol (7-O-methylaromadendrin), two lignans (pinoresinol and matairesinol) and six diterpenic acids/alcohol (19-acetoxy-13-hydroxyabda-8(17),14-diene, totarol, 7-oxodehydroabietic acid, dehydroabietic acid, communic acid and isopimaric acid) were isolated from the hydroalcoholic extract of a Brazilian Brown Propolis and characterized by NMR spectral data analysis. The volatile fraction of brown propolis was characterized by CG-MS, composed mainly of monoterpenes and sesquiterpenes, being the major α-pinene (18.4 %) and β-pinene (10.3 %). This propolis chemical profile indicates that Pinus spp., Eucalyptus spp. and Araucaria angustifolia might be its primary plants source. The brown propolis displayed significant activity against Plasmodium falciparum D6 and W2 strains with IC₅₀ of 5.3 and 9.7 μg/mL, respectively. The volatile fraction was also active with IC₅₀ of 22.5 and 41.8 μg/mL, respectively. Among the compounds, 1-O,2-O-digalloyl-6-O-trans-p-coumaroyl-β-D-glucopyranoside showed IC₅₀ of 3.1 and 1.0 μg/mL against D6 and W2 strains, respectively, while communic acid showed an IC₅₀ of 4.0 μg/mL against W2 strain. Cytotoxicity was determined on four tumor cell lines (SK-MEL, KB, BT-549, and SK-OV-3) and two normal renal cell lines (LLC-PK1 and VERO). Matairesinol, 7-O-methylaromadendrin, and isopimaric acid showed an IC₅₀ range of 1.8–0.78 μg/mL, 7.3–100 μg/mL, and 17–18 μg/mL, respectively, against the tumor cell lines but they were not cytotoxic against normal cell lines. The crude extract of brown propolis displayed antimicrobial activity against C. neoformans, methicillin-resistant Staphylococcus aureus, and P. aeruginosa at 29.9 μg/mL, 178.9 μg/mL, and 160.7 μg/mL, respectively. The volatile fraction inhibited the growth of C. neoformans at 53.0 μg/mL. The compounds 3-hydroxy-4-methoxybenzaldehyde, 3-methoxy-4-hydroxypropiophenone and 7-oxodehydroabietic acid were active against C. neoformans, and caffeic and communic acids were active against methicillin-resistant Staphylococcus aureus.     

Green and Red Brazilian Propolis: Antimicrobial Potential and Anti-Virulence against ATCC and Clinically Isolated Multidrug-Resistant Bacteria

Brazilian green and red propolis stand out as commercial products for different medical applications. In this article, we report the antimicrobial activities of the hydroalcoholic extracts of green (EGP) and red (ERP) propolis, as well as guttiferone E plus xanthochymol (8) and oblongifolin B (9) from red propolis, against multidrug-resistant bacteria (MDRB). We undertook the minimal inhibitory (MIC) and bactericidal (MBC) concentrations, inhibition of biofilm formation (MICB₅₀), catalase, coagulase, DNase, lipase, and hemolysin assays, along with molecular docking simulations. ERP was more effective by displaying MIC and MBC values <100 μg mL⁻¹. Compounds 8 and 9 displayed the lowest MIC values (0.98 to 31.25 μg mL⁻¹) against all tested Gram-positive MDRB. They also inhibited the biofilm formation of S. aureus (ATCC 43300 and clinical isolate) and S. epidermidis (ATCC 14990 and clinical isolate), with MICB₅₀ values between 1.56 and 6.25 μg mL⁻¹. The molecular docking results indicated that 8 and 9 might interact with the catalase's amino acids. Compounds 8 and 9 have great antimicrobial potential.     

Uncovering Biological Application of Brazilian Green Propolis: A Phenotypic Screening against Schistosoma mansoni

The chemotherapy of schistosomiasis remains centered in the use of praziquantel, however, there has been growing resistant parasites to this drug. Thus, the aim of this work was to evaluate in vitro schistosomicidal activity of the hexanes/dichloromethane 1 : 1 extract of Brazilian green propolis (Pex), as well as its major isolated compounds artepillin C, caffeic acid, coumaric acid and drupanin against Schistosoma mansoni. The Pex was active by displaying an IC₅₀ value of 36.60 (26.26–51.13) μg mL^?1 at 72 h against adult worms of S. mansoni. The major isolated compounds were inactive with IC₅₀ values >100 μM, however, the combination of the isolated compounds (CM) in the same range found in the extract was active with an IC₅₀ value of 41.17 (39.89–42.46) μg mL^?1 at 72 h. Pex and CM induced alteration in the tegument of S. mansoni, and caffeic acid caused alteration in egg's maturation. Pex displayed in vitro activity against adult worms’ and eggs’ viability of S. mansoni, which opens new perspectives to better understand the synergistic and/or additive effects promoted by both Pex extract and CM against schistosomiasis features.     

2008~2012年蜂胶化学成分研究进展

近年来,蜂胶的化学成分成为了国内外蜂胶研究的热点。本文对2008~2012年国内外从蜂胶中分离鉴定出的新化学成分进行了分类汇总,给出了每种化学成分的名称以及蜂胶的地理来源,以期为蜂胶化学成分的研究、胶源植物的确定以及质量控制提供理论依据。本文共列出144种蜂胶中的新成分,其中,类黄酮化合物68种、萜类化合物38种、酚酸类化合物38种。     

Bromeliaceae diversity and conservation in Minas Gerais state,Brazil

Field work and data from herbaria collections (2686 records) representing 283 taxa (265 species and 18 infraspecific taxa) of Bromeliaceae occurring at Minas Gerais state, southeastern Brazil, were analyzed in order to obtain distribution and diversity information, and to determine IUCN (The World Conservation Union) conservation status for each taxon. A map containing 1° × 1° grid cells was used to identify priority areas for new research collections, areas of high species diversity, and Bromeliaceae conservation status. A clear decrease in Bromeliaceae diversity is observed between the eastern and the western portions of Minas Gerais, and low floristic similarities were found between neighboring grid cells. The rocky mountains of Cadeia do Espinhaço are considered the most important area for Bromeliaceae endemics. From the 283 taxa of Bromeliaceae that occur at Minas Gerais, 118 (42%) are considered threatened, and 124 taxa (44% of the total) do not occur inside any protected area. The region of the Quadrilátero Ferrífero in the southern portion of the Cadeia do Espinhaço is the most threatened, and urgent strategies for conservation of this rich Bromeliaceae flora are needed. Northeastern Minas Gerais, particularly the rocky outcrops or inselbergs located in the Jequitinhonha and Mucuri rivers drainage basins need additional collection efforts and conservation actions focused on these saxicolous taxa.     

不同来源中国蜂胶水提物的化学组成及抗氧化性

本文研究了季节和地理来源对中国蜂胶水提物的化学成分和抗氧化性的影响。分别采集了来自山东、湖北和海南三个省份不同季节的蜂胶,采用改进的提取方法,得到蜂胶水提物(Water extract of propolis,WEP)。分光光度法测定了WEP的主要化学组成,普鲁士兰法和清除DPPH自由基活性法评价了WEP的抗氧化性。研究结果表明,中国WEP含有总酚和总黄酮等化学成分,具有较好的抗氧化性。季节对中国WEP化学成分和抗氧化性有显著影响。山东和湖北WEP化学成分比较接近,但同海南WEP有显著差异,抗氧化活性也显著强于海南WEP。不同产地和不同采收季节的中国WEP存在化学组成及生物活性多样性,在生产实践中需注意这一特性。     

Congenital neosporosis in goats from the State of Minas Gerais, Brazil

Congenital Neospora caninum infection was diagnosed in two Saanen goat kids from two distinct herds with a history of abortion and weak newborn goat kids in the Southern region of the State of Minas Gerais, Brazil. The first kid was weak at birth, had difficulty to rise and was unable to nurse. Gross lesions of porencephaly and hydrocephalus ex vacuo were seen. Multifocal necrosis, gliosis and non-supurative encephalitis were observed in the brain. Several parasitic cysts with a thick wall that reacted strongly only with polyclonal antiserum to Neospora caninum were seen in the cerebral cortex, brain stem and cerebellum. The second kid was born from a Neospora caninum seropositive mother that aborted in the last pregnancy. It was born without clinical signs. The diagnosis of neosporosis was based on antibody titer of 1:800 to N. caninum by indirect fluorescence antibody test obtained from blood collected before the goat kid ingested the colostrum and Neospora caninum DNA was detected by polymerase chain reaction and sequenced from placenta. This is the first report of neosporosis in goats in the southeast region of Brazil.     

Chemical constituents of Brazilian Propolis from the state of Bahia and their growth inhibitory activities against cancer cells

A novel 2-phenoxychromone (1) and five known flavones (2-6) were isolated from northeastern Brazilian propolis in the state of Bahia. The chemical structures of these six compounds were determined by spectroscopic investigations and single-crystal X-ray analysis. The isolated compounds showed growth-inhibitory activities, in varying degrees, against human tumor cell lines. This is the first report on the discovery of a novel 2-phenoxychromone from propolis.     

Entomopathogenic nematode distribution and edaphoclimatic conditions in the Cerrado of Minas Gerais,Brazil

Based on the high diversity of Brazilian fauna and flora, edaphoclimatic conditions in the Cerrado of Minas Gerais, and the situation of utilization of EPNs in Brazil, a survey was conducted in order to relate the presence of these organisms with the physical and chemical attributes of the soil, combined with precipitation. To this end, soil samples were collected in areas with diversified vegetation types in Monte Carmelo, MG, at Juliana Farm. The samples were obtained every 15 days for 6 months. From each spot, soil samples (about 500 g) were collected for soil moisture characterization, nematode isolation and determination of pH, organic matter, potassium (K), calcium (Ca), magnesium (Mg), phosphorus (P), potential acidity (H + Al) and precipitation data (mm). Three populations of entomopathogenic nematodes of the Heterorhabditis amazonensis species were isolated in Cerrado stricto sensu and Gallery forest areas. The occurrence H. amazonensis could not be considered restricted to specific soil condition, as organic matter, humidity, pH, Ca, K, Mg and H + Al, especially considering the organic matter and K values, which had variable levels between the places of collection. The p values of the positive soil samples were at a lower level than the mean of the Gallery forest and Cerrado, and at the same level as maize and pasture area. The soil moisture in the Cerrado area increased with the higher values of precipitation; however, in the Gallery forest area this association was not observed. Also, the nematodes were isolated when the temperature began to decrease.     

Areas of endemism in the Espinhaço Range in Minas Gerais, Brazil

The Espinhaço Range, a mountain chain located in the Brazilian states of Minas Gerais and Bahia, contains one of the richest floras in Brazil, with a high frequency of endemic species. Since 2005 it is designated as UNESCO biosphere reserve and is situated at the joint border of two global hotspots for biodiversity conservation. Endemic species with congruent occurrence patterns were identified in order to establish areas of higher endemism within the Espinhaço Range. Taxonomic reviews were analyzed in order to identify endemic taxa and a dataset was elaborated containing 1765 records from 178 endemic species of vascular plants, representing 17 families and including the geographic coordinates for each record. Two maps were produced showing species richness and collection effort in 15′ quadrats. The congruent occurrences were identified and a third map was provided, delimiting 10 candidate areas of endemism for a “parsimony analysis of endemicity” (PAE). One most parsimonious cladogram is then retrieved, evidencing three major clades corresponding to the northern, central and southern portions of the Espinhaço, in addition to four subclades included into the central clade. We finally identified six major areas of endemism. Furthermore, there is a strong correlation between species richness and collector effort, revealing which areas are in need of further field inventories.     

Clinical, epidemiological and outcome features of patients with cryptococcosis in Uberaba, Minas Gerais, Brazil

Nearly one million of cryptococcosis cases occur yearly around the world, involving mainly HIV-infected patients who are not receiving antiretroviral therapy (ART) or present poor adherence. This study aims to evaluate epidemiological, clinical and outcome aspects of patients with cryptococcosis from 1998-2010. Patients were prospectively recruited, and their medical and laboratory records were reviewed. A total of 131 cases were included, and of these, 119 (90.83%) had AIDS, 4 received a renal transplant, 2 presented systemic lupus erythematosus and 6 (4.6%) were apparently immunocompetent. Ninety-one (69.46%) were men, and the median age was 38.7?years. Cryptococcal meningitis (CM) was diagnosed in 103 (78.62%), whereas 28 (21.38%) had cryptococcal infection in other sites. Of patients with CM, 94 (91.26%) had AIDS being cryptococcosis the first defining illness in 61 (64.9%), while 37 (60.65%) of them presented simultaneously both diagnosis. Headache, altered mental status, papilledema and seizures at admission were significatively associated with a poor outcome. Of 163 different isolates, 155 (95.09%) were Cryptococcus neoformans and eight (4.88%) Cryptococcus gattii. Antifungal therapy was warranted in 8 (87.4%) patients with CM, but 46 (51.1%) died during the first days or weeks. Of 28 patients without CM, 21?(75%) received treatment, but 6 (28.6%) died. The poor outcome among this case series was similar to that reported from other developing countries, but it is paradoxal in Brazil where the ART is at free disposal in the public health services. Despite, at least 60-70% of patients present advanced immunosuppression when they receive the AIDS diagnosis.     

LC-ESI-MS/MS Chemical Characterization,Antioxidant and Antidiabetic Properties of Propolis Extracted with Organic Solvents from Eastern Anatolia Region

Geographic conditions (altitude, climate, and local flora) lead to significant differences in the chemical composition of propolis. Therefore, more research is needed for propolis in different geographical regions. So, the aim of this study was to evaluate the phenolic profile, total phenolic content, antioxidant, and antidiabetic properties of Pülümür propolis from Turkey. Methanol (MeOH), chloroform (CHCl₃), and hexane extracts of propolis were analyzed. LC-ESI-MS/MS analysis of the extracts showed that the most abundant phenolic compound is caffeic acid in the MeOH extract (2943.12±11.12 μg phenolics/g extract), while on the other hand, CHCl₃ extract had the highest total phenolic content (125.75±1.02 mg GAE/g extract). Antioxidant activity was measured using ABTS and DPPH assays, whereas CHCl₃ extract (IC₅₀=6.35±0.11 and 28.84±0.10 μg/mL, respectively) and MeOH extracts (IC₅₀=5.04±0.07 and 28.80±0.09 μg/mL, respectively) showed relatively high antioxidant activity. The MeOH extract showed better antidiabetic activity than the standard compound, acarbose (IC₅₀=0.544 and 0.805 mg/mL, respectively).     

天然棕色棉纤维色素光谱学特性及其化学结构初步推断

在纯化棕色棉纤维色素的基础上,测定了天然棕色棉纤维色素的紫外-可见光谱（UV光谱）、红外光谱（IR光谱）以及分析了pH值、浓度、诊断试剂对天然棕色棉纤维色素UV光谱的影响,发现天然棕色棉纤维色素UV光谱随着pH值和浓度不同而变化,诊断试剂对纤维色素UV光谱也有影响,但并不符合黄酮类化合物对诊断试剂的反应规律,红外光谱的主要吸收波数是3554、3477、3414、3289、1638、1618、1386、1073、617、478cm^-1。结合化学性质鉴定,以及和已知的白色棉种皮色素比较。初步推断棕色棉色素的化学结构,认为棕色棉纤维中的色素是由单宁物质氧化形成的醌类化合物。最后解释了棕色棉纤维具有颜色是因为棉纤维发育后期缩合单宁接触空气后氧化的结果。     

Caseous lymphadenitis in sheep flocks of the state of Minas Gerais,Brazil: Prevalence and management surveys

Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, which is a serious, economically important problem for sheep production. We examined the seroprevalence of infection by C. pseudotuberculosis and possible risk factors associated with caseous lymphadenitis in sheep herds of the state of Minas Gerais, Brazil. Samples were collected from 642 sheep from 97 farms. Sera of all of the sheep were tested with ELISA for antibodies against C. pseudotuberculosis. A questionnaire was applied to gather data on the farm, the sheep herd, the farmer, and individual animal data (breed, sex and age). This is the first sero-epidemiological survey for caseous lymphadenitis in sheep herds in Minas Gerais. We found a high real prevalence, much higher than that suggested from information obtained with the questionnaire, which points to the scarcity of vaccination against caseous lymphadenitis in the sample evaluated. Only a small proportion of the farmers declared that cases of this disease were present in their flocks. The frequency of seropositive sheep varied significantly with breed (χ² test, P = 0.021). Age group also significantly affected the percentage of seropositivity (χ² test, P = 0.049), the highest frequency being found in adult animals (more than 12 months old), when compared to the 5–12 months old group (χ² test, P = 0.021). The prevalence of infection with C. pseudotuberculosis in sheep in the state of Minas Gerais was estimated to be 70.9% (95% confidence interval (CI): 64.7–77.0%) and the prevalence of infected flocks being 95.9% (95% CI: 89.8–98.9%). We concluded that C. pseudotuberculosis infection is widely disseminated in sheep flocks in Minas Gerais and that caseous lymphadenitis control and eradication programs are necessary in this state.     

The unique and endangered Campo Rupestre vegetation and protected areas in the Iron Quadrangle,Minas Gerais,Brazil

The Iron Quadrangle (IQ) in Brazil is one of the world's largest reserves of iron ore, and besides its economic importance, the region also plays a key role in nature conservation with the endemic Campo Rupestre vegetation. Despite a large number of protected areas (PAs) offering refuge for the remaining Campo Rupestre vegetation in the IQ, studies on the current situation of these natural remnants, and the real functionality of these PAs are scarce. This study aimed to map the remaining natural phytophysiognomies of the IQ, evaluating the proportion of Campo Rupestre on ferruginous and non-ferruginous substrates in PAs. We applied the object-oriented classification using Sentinel 2A and 2B satellite images, vegetation indexes, iron oxide index and terrain attributes, with field-collected validation points. We identified thirteen types of land use and cover, of which five represented natural phytophysiognomies. The most common phytophysiognomies in the IQ are Seasonal Forests and Cerrado (all types included), accounting for more than 60% of the total area. Campo Rupestre on ferruginous and non-ferruginous substrates accounts for 5.4% of the total mapped area only. On the other hand, about 42.5% of the Campo Rupestre (ferruginous or not) are protected and correspond to 17.7% of the PAs. We show that an appropriate mapping of the phytophysiognomies in the IQ region is key for improving landscape ecology knowledge and future conservation planning, especially of the endangered Campo Rupestre vegetation.     

Diel migration and vertical distribution of Cladocera in Lake D. Helvecio (Minas Gerais,Brazil)

Six genera of Clad ocera (Diaphanosoma, Daphnia, Ceriodaphnia, Moina, Bosmina, Bosminopsis), each of them usually with only one species were found in Lake D. Helvecio, a natural valley lake located in the eastern part of Brazil. Diurnal migratory movements of the organisms observed in this lake showed a different pattern in different species. Closely related species, which explore the same food source, live in different layers, thus avoiding interspecific competition. The migratory behaviour of the species was studied mainly in relation to temperature and oxygen distribution in the lake. Thus, analyses were made in the summer (January, 1978) when a strong stratification occurs with the establishment of a thermocline and an oxycline. Comparisons were made also with the data obtained in winter (July, 1978), when a complete mixing of water occurs.     

Variability of Meloidogyne exigua on Coffee in the Zona da Mata of Minas Gerais State,Brazil

Minas Gerais is the major coffee-producing state of Brazil, with 28% of its production coming from the region of Zona da Mata. Four major species of root-knot nematode attacking coffee (Meloidogyne incognita, M. paranaensis, M. coffeicola, and M. exigua) have been reported from Brazil. To determine the variability in Meloidogyne spp. occurring in that region, 57 populations from 20 localities were evaluated for morphological, enzymatic, and physiological characteristics. According to the perineal pattern, all the populations were identified as M. exigua; however populations from the municipality of São João do Manhuaçu exhibited patterns very similar to M. arenaria. The identity of all the populations was confirmed by the phenotypes of esterase, malate dehydrogenase, superoxide dismutase, and glutamate-oxaloacetate transaminase. Thirteen populations (22.8%) showed the typical one-band (E1) esterase phenotype, whereas the others (77.2%) had a novel two-band phenotype (E2). No intraspecies variability was found in any population. All populations were able to reproduce on tomato, pepper, beans, cacao, and soybean. Reproduction was greater on tomato and pepper than on coffee seedlings, the susceptible standard.     

Checklist and one new species of Bromeliaceae from Pico do Itambé, Minas Gerais,Brazil

A checklist of the 11 genera and 16 species of Bromeliaceae from the Parque Estadual do Pico do Itambé in Santo Antônio do Itambé and Serra Azul de Minas, Minas Gerais, south‐eastern Brazil, is presented. Bromelioideae is the most diverse subfamily and is concentrated in forested areas. Tillandsioideae and Pitcairnioideae are most abundant in elevated areas of campo rupestre vegetation. Vriesea medusa Versieux, a new night‐blooming species belonging to section Xiphion (E.Morren) E.Morren ex Mez, was found in the area and is described and illustrated. It appears to be an endemic species distinguished from Vriesea nanuzae Leme by a combination of the usually larger dimensions in the rosette, the curved inflorescence which has lateral peduncles bearing up to five sterile bracts, the larger number of flowers, floral bracts and sepals that become dark purplish‐brown towards the apex and the coloration of the leaves. Dyckia glandulosa L.B.Sm. & Reitz and Vriesea densiflora Mez, previously poorly known, were rediscovered and are depicted in their habitat for the first time. Vriesea procera var. tenuis L.B.Sm. is a new record for the state. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 709–715.