Antifungal rosane diterpenes and other constituents of Hugonia castaneifolia

The rosane diterpenoids hugorosenone [3beta-hydroxyrosa-1(10),15-dien-2-one], 18-hydroxyhugorosenone and 18-hydroxy-3-deoxyhugorosenone, and 12-hydroxy-13-methylpodocarpa-8,11,13-trien-3-one were isolated as antifungal constituents of H. castaneifolia Engl. root bark, together with the previously reported di-podocarpanoids hugonone A and hugonone B that were weakly active, and 1(10),15-rosadiene-2beta,3beta-diol (hugorosenol), 4alpha-methoxyhimachal-10-en-5beta-ol (hugonianene B) and 2-hydroxyhenpentacont-2-enal, and the known compounds tetracosyl-(E)-ferrulate and caryophyllene oxide, all of which were inactive. Hugorosenone also exhibited activity against Anopheles gambiae mosquito larvae. Structural determination was achieved based on spectroscopic data.     

Effect of lactoferrin on enteric pathogens

Much has been learned in recent years about the mechanisms by which breastfeeding improves child health and survival. However, there has been little progress in using these insights to improve pediatric care. Factors that are important for protecting the breast fed infant might be expected to decrease the adverse effects of weaning on diarrhea, growth, and development. Lactoferrin, an iron-binding protein with multiple physiological functions (anti-microbial, anti-inflammatory, and immunomodulatory), is one of the most important proteins present in mammalian milk. Protection against gastroenteritis is the most likely biologically relevant activity of lactoferrin. Multiple in vitro and animal studies have shown a protective effect of lactoferrin on infections with enteric microorganisms, including rotavirus, Giardia, Shigella, Salmonella and the diarrheagenic Escherichia coli. Lactoferrin has two major effects on enteric pathogens: it inhibits growth and it impairs function of surface expressed virulence factors thereby decreasing their ability to adhere or to invade mammalian cells. Thus, lactoferrin may protect infants from gastrointestinal infection by preventing the attachment by enteropathogens in the gut. Recently several clinical trials in children have started to address this issue. Whether lactoferrin can prevent a significant portion of diarrheal disease remains to be determined.     

Influence of root temperature on growth of Pinus sylvestris,Fagus sylvatica,Tilia cordata and Quercus robur

One-year-old tree seedlings were incubated in a greenhouse from April to July, under natural daylight conditions, with their root systems at constant temperatures of 5, 10, 15, 20, 25, 30 and 35 °C and with the above ground parts kept at a constant air temperature of 18–20 °C. The course of height growth, total mass increment, root, shoot and leaf weight as well as leaf areas were measured. The results indicate that clear differences exist in the optimal root zone temperatures for various growth parameters in different tree species. Pinus sylvestris had a maximal height increment at about 5–10 °C and maximal total mass increment at 15 °C root temperature. In contrast, the optimum for Quercus robur was at 25 °C. Tilia cordata and Fagus sylvatica had their optima for most growth parameters at 20 °C. The root temperature apparently indirectly influenced photosynthesis (dry weight accumulation) and respiration loss. From the observed symptoms and indications in the literature it seems probable that a change in hormone levels is involved as the main factor in the described effects. Variation of root temperature had only an insignificant effect on bud burst and the time at which the shoots sprouted. Apparently species of northern origin seem to have lower root temperature optima than those of more southern origin. This is to be verified by investigation of other tree species.     

Notes on Quercus ilex L. in Liguria (NW Italy)

The climatic characteristics of the Liguria region have been outlined by means of raw data and derived indices (water balance related to potential evapotraspiration, Rivas-Martinez's index of mediterraneity and thermicity, De Martonne's index of dryness, and Emberger's pluviothermic quotient and index of summer dryness). Their interpretation suggests that Liguria is a boundary region between two different climatic areas: the Mediterranean and that of Central Europe.The distribution of Quercus ilex communities in Liguria have suffered the heavy consequences of human activity on the coastal belt of the region. There is, however, sufficient evidence to show that their presence is closely linked to climatic conditions. Quercus ilex communities are absent from the western coastal belt where the annual water balance is below –50 mm and high mean temperatures occur. They are also absent from marly limestone dominated areas where annual water balance is below 300 mm. Quercus ilex woods show a preference for water balance values between 0 and 600, with the better developed forests being found in central and eastern coastal Liguria.The Ligurian Quercus ilex woods are ascribed to Quercetum ilicis Br.-Bl. 1915. Given that the presence within their floristic composition of a group of species of Querco-Fagetea (more species in eastern stands, few in western ones) is an almost constant characteristic, the subass. fraxino-ostryetosum Mariotti 1984 can be retained for many of them. Quercus ilex is not present in the initial succession stages of Mediterranean Liguria vegetation. It can be found in later stages when the vegetation is more fully developed towards a maquis type structure. In this case it tends to become dominant and the community develops (if no fire or other human interference occurs) into a Quercus ilex forest. Frequently, this succession takes place under an old tree layer cover (generally Pinus sp.), a remanant of man's past management of the vegetation in the area.     

Near on-line detection of enteric bacteria using lux recombinant bacteriophage

The potential for a revolution in microbial testing can be perceived with the near on-line detection of indicator microorganisms. By definition, these are microorganisms present in significant numbers within a food which, while not pathogenic, can be related through increasing count to the increased probability of pathogen contamination. We have used recombinant lux+ bacteriophage to detect enteric indicator bacteria without recovery or enrichment in 50 min, provided that they are present at levels greater than 10(4) g-1 or cm-2. After a 4-h enrichment, samples having enteric counts of 10 g-1 or cm-2 can be distinguished from background.     

The efficiency of nitrogen retranslocation from leaf biomass in Quercus ilex ecosystems

Nitrogen retranslocation from senescing leaves represents a crucial adaptation by tree species towards a more efficient use of this nutrient. As a result, this part of the nitrogen cycle has received increasing attention in recent years. However, there remain strong discrepancies with respect to the factors responsible for interspecific differences in the efficiency of this process.In the present work the seasonal pattern of leaf growth and the movement of nitrogen in leaves have been studied in a series of Quercus ilex plots with different levels of rainfall and soil quality in central-western Spain, as well as in 20 other woody species typical of this area. The percentage of nitrogen retranslocated was estimated from the difference between the maximum mass of nitrogen stored in the leaf biomass and the amount of this nutrient returned annually to the soil through leaf fall. Q. ilex appears as one of the least efficient species in the Mediterranean region in the recovery of nitrogen from senescing leaves (29.7% of the maximum pool). Furthermore, the older leaves of Q. ilex do not show the cycles of nitrogen withdrawal during new flushes of shoot growth, such as occurs in Pinus spp. This suggests that older leaves in Q. ilex do not play an important role as nitrogen storage organs.     

The inhibitory activity of Lactobacillus spp. isolated from breast milk on gastrointestinal pathogenic bacteria of nosocomial origin

Milk acts as a mean for transporting many essential substances from the mother to the child. In human beings, milk includes several predominant bacteria, such as staphylococci, streptococci, micrococci, lactobacilli, enterococci, lactococci and bifidobacteria. Besides, its intake favors the predominance of bifidobacteria and lactobacilli in the child’s intestinal microbiota. The present work explores the isolation and selection of lactobacilli strains with probiotic potential, focusing in their degree of hydrophobicity and antagonism against important gastrointestinal nosocomial pathogens. 98 lactobacilli were isolated from 48 breast milk samples, with most strains belonging to the obligately homofermentative group (36.7%). 63% of the isolated strains showed a high degree of hydrophobicity when tested on three solvents and were selected for detecting antimicrobial activity against gastrointestinal pathogens, including Escherichia coli, Shigella spp, Pseudomonas spp and Salmonella spp strains. When applying the agar diffusion test, many isolated strains presented inhibitory activity against pathogenic strains. We observed that: Salmonella enteriditis was the most inhibited pathogen, and the strains with the most inhibitory power were AR2 and O1 (both highly hydrophobic lactic acid bacteria), which showed an opposing effect against all nosocomial pathogens tested. Although more in vitro, in vivo or clinical data would be needed before any conclusion on the probiotic properties of the strains can be drawn, our results demonstrate that some of the tested strains may have good probiotic potential for their inclusion in products targeting infants.     

Redheadia quercus gen. et sp. nov., the teleomorph of Mycopappus quercus, the frosty mildew fungus in Quercus acutissima

The teleomorph of Mycopappus quercus causing frosty mildew in Quercus acutissima is described as a new genus and species, Redheadia quercus, in the Sclerotiniaceae. Apothecia sprout from sclerotia on the fallen infected leaves kept for 10 months at 5°C and subsequent incubation at 15°C under diffused room light. Typical zonate lesions and multicellular propagules of M. quercus are produced on Q. acutissima, by mycelial inoculation using an isolate from a single ascospore, confirming the teleomorph–anamorphic connection. No significant differences are observed between cultured colonies of isolates from the ascospore and those from the propagule. Sclerotia and microconidia of the fungus are produced on culture media.     

Occurrence of airborne enteric bacteria in Mexico city

Summary An investigation of microbial air quality in the area of the National Autonomous University of Mexico, located in the southern part of Mexico City, was conducted for one year. Ambient outdoor concentrations and size distribution of airborne bacteria were measured, 130 samples were taken at noon, using an Andersen 6 stage sampler, located 2 m above ground level. Concentration ranges and colony-forming units per cubic meter of air (CFU m^–3) found, were as follows:14 to 12999 for total bacteria, No growth (NG) to 55 for coliform bacteria, NG to 11 for fecal coliform and NG to 10 for fecal Streptococci.Bacteria associated with the potentially respirable fraction (0.65 to 4.7 µm) averaged 37% and 9% for total bacteria and coliform bacteria respectively. In 23% of the samples, coliform bacteria were recovered, with higher incidences during dry season. The most common of these were:Escherichia coli (15%), followed bySerratia (13%) andEnterobacter (10%),The total bacteria correlated significatively (p<0.05) with the following parameters: particulate matter smaller than 10 µm (PM₁₀) (r=0.40), total suspended particulates (TSP) (r=0.26), daily variation of temperature (r=0.18), and vapor pressure (r=–0.16). These relationships indicate that fecal soil pollution could affect air quality with potential health risks.     

Effect of cleaning and thinning on height growth and girth increment in holm oak coppices (Quercus ilex L.)

This article studies the influence of cleaning or thinning intensity and of coppice age at the time of silvicultural treatment, on the growth of shoots remaining on the stool, in Quercus ilex coppices.Two experimental designs were used to answer these questions. The first studied the effect of two cleaning intensities on 4, 8, 15, 20, and 25 year old coppices. The second studied the effect of 4 thinning intensities (26%, 42%, 58%, or 78% removal of the initial basal area) on 43 and 57 year old coppices. In both cases, control plots undergoing no silvicultural treatment were maintained for each age class.The results, 4 and 5 years after silvicultural treatment, concerned girth increment and height growth of coppice shoots.Cleaning or thinning consistently had a positive influence on girth increment. In young coppices, less than 25 years old, mean annual girth increment was approximately 6 mm in the controls and double that under heavy cleaning. In older coppices, girth increment was lower in controls (3 mm per year) but the effect of thinning was very positive and, under very heavy thinning, growth was more than 11 mm per year.Cleaning and thinning often modify height growth. This effect was found to be either positive or negative and no general trend could be defined which was valid for all the age classes.Silvicultural treatments also had a positive effect on the appearance and growth of epicormic shoots on the holm oak trunks.The results were also analysed at the stool level. The number or percentage of shoots removed had a positive effect on the individual growth of selected shoots.     

Development of a prototype wound dressing technology which can detect and report colonization by pathogenic bacteria

A new methodology for detecting the microbiological state of a wound dressing in terms of its colonization with pathogenic bacteria such as Staphylococcus aureus or Pseudomonas aeruginosa has been developed. Here we report how stabilized lipid vesicles containing self-quenched carboxyfluorescein dye are sensitive to lysis only by toxins/virulence factors from P. aeruginosa and S. aureus but not by a non-toxic Escherichia coli species. The development of the stabilized vesicles is discussed and their response to detergent (triton), bacterial toxin (α-hemolysin) and lipases (phospholipase A₂). Finally, fabrics with stabilized vesicles attached via plasma deposited maleic anhydride coupling are shown visibly responding to S. aureus (MSSA 476) and P. aeruginosa (PAO1) but not E. coli DH5α in a prototype dressing.     

Lactic acid bacteria in fermented foods in Thailand

Traditional fermented foods (fish, meat and vegetable products), produced by many different processes, are eaten in many parts of Thailand. Lactic acid bacteria are responsible for the souring and ripening of these foods. Homofermentative strains of Lactobacillus pentosus, L. plantarum and Pediococcus pentosaceus are dominant in foods with low salt concentrations whereas P. halophilus strains are present in foods containing high salt. Strains of Lactobacillus sake, other Lactobacillus spp., P. acidilactici and P. urinaeequi are frequently found. Heterofermentative strains of L. brevis, L. confusus, L. fermentum, L. vaccinostercus, other Lactobacillus spp., and of Leuconostoc spp. are distributed as minor bacteria and strains of Staphylococcus, Enterococcus and Halobacterium are occasionally isolated.S. Tanasupawat is with the Department of Microbiology, Faculty of Pharmaceutical Sciences. Chulalongkorn University, Bangkok 10330, Thailand; K. Komagata is with the Department of Agricultural Chemistry, Tokyo University of Agriculture, Sakuragaoka 1-1-1, Setagaya-ku, Tokyo 156, Japan.     

Quantification of factors affecting the probability of development of pathogenic bacteria,in particularClostridium botulinum,in foods

Summary The success of a preservation method with respect toClostridium botulinum can be measured by its effect on the probability that a single spore would result in growth and formation of toxin. In canned, low-acid foods, the minimum heat-process is designed to reduce the probability of survival of a single, heat-resistant spore ofC. botulinum by a factor of 10¹². In some foods, safety depends on the combination of inactivation and inhibition ofC. botulinum. The degree of protection (Pr) can be expressed asPr=Ds+In, whereDs is the decimal destruction of spores ofC. botulinum andIn is the decimal inhibition. A similar approach can be used in the case of other bacteria.Pr=log 1/P, whereP=the probability that an individual spore or bacterium will survive and result in growth.P can be estimated as the number of spores or bacteria that survive and initiate growth in a culture medium or food divided by the number of viable spores or bacteria inoculated into the medium or food. The effects of combinations of preservative factors can be measured by their effect onP at a stated temperature for a stated time. In experiments to determine the effects of preservative factors on an anaerobic bacterium such asC. botulinum it is essential that oxygen should be eliminated, unless it is controlled as an inhibitory factor. Thus experiments in culture media should be done under strictly anaerobic conditions at a known, low redox potential. The results of experiments to determine the effects of preservative factors onP after a series of incubation times can be modeled by methods similar to those used to model the effect of preservative factors on rate of growth and on lag period. Experiments to determine the effect of preservative factors on the probability of growth from a single spore or bacterium ofC. botulinum are discussed. A few reports of similar experiments with other bacteria have been published and are described. This approach has the advantage that it takes account of the inoculum level of bacteria.Mention of brand of firm names does not constitute an endorsement by the US Department of Agriculture over others of a similar nature not mentioned.     

Detection and features of wetwood in Quercus mongolica var. grosseserrata

To elucidate the location of water in wetwood, variations in moisture content within a tree of Quercus mongolica var. grosseserrata were observed macroscopically by determination of moisture content and by soft x-ray photography. Then the typical wetwood, which had been precisely located by such macroscopic analysis, was examined further by cryo-scanning electron microscopy. The moisture content varied considerably within the tree trunk, and typical wetwood was detected in limited regions of the heartwood. The distribution of the wetwood did not always correspond to the discoloration of the wood. In the wetwood, almost all the lumina of vessels and fibers were filled with water, while the lumina of ray and axial parenchyma cells were often cavitated. Intercellular spaces produced by deformation of cell walls and radial shakes were filled with water. In addition, tyloses were collapsed and torn in the wetwood, and they did not block the vessel lumina. These results suggest the possibility that vessels and fibers might serve as effective pathways for the accumulation of water in the wetwood.     

CvL, a lectin from the marine sponge Cliona varians: Isolation, characterization and its effects on pathogenic bacteria and Leishmania promastigotes

CvL, a lectin from the marine sponge Cliona varians was purified by acetone fractionation followed by Sepharose CL 4B affinity chromatography. CvL agglutinated papainized treated human erythrocytes with preference for type A erythrocytes. The lectin was strongly inhibited by monosaccharide d-galactose and disaccharide sucrose. CvL is a tetrameric glycoprotein of 28 kDa subunits linked by disulphide bridges with a molecular mass of 106 kDa by SDS-PAGE and 114 kDa by Sephacryl S300 gel filtration. The lectin was Ca2+ dependent, stable up to 60 degrees C for 60 min, with optimum pH of 7.5. CvL displays a cytotoxic effect on gram positive bacteria, such as Bacillus subtilis and Staphylococcus aureus. However, CvL did not affect gram negative bacteria, such as Escherichia coli and Pseudomonas aeruginosa. Leishmania chagasi promastigotes were agglutinated by CvL up to 2(8) titer. These findings are indicative of the physiological defense roles of CvL and its possible use in the antibiosis of bacteria and protozoa pathogenic.     

The association of algicidal bacteria and raphidophyte blooms in South Carolina brackish detention ponds

Over the past 5 years, raphidophyte blooms have been frequently observed along the South Carolina coastal zone. During the 2002, 2003, and 2004 sampling seasons, we investigated temporal fluctuations of algicidal bacteria abundance against raphidophycean flagellates (Heterosigma akashiwo, Chattonella subsalsa, and Fibrocapsa japonica) using the microplate most probable number (MPN) method in three Kiawah Island brackish stormwater detention ponds (K1, K2, and K75). Local axenic isolates of H. akashiwo, C. subsalsa, and F. japonica were obtained and their susceptibility to algicidal bacteria tested. A total of 195 algicidal bacterial strains were isolated from raphidophyte blooms in the study ponds, and 6 of them were identified at the genus level, and the taxonomic specificity of their algicidal activity was tested against local (pond) and nonlocal isolates of raphidophytes (3 species, 10 total strains). In the ponds, a consistent association was found between raphidophyte bloom development and an increase in bacteria algicidal to the bloom species. In 12 of 15 cases, bloom decline followed the increase in algicidal bacteria to maximum abundances. Although variability was found in the taxonomic specificity of the algicidal bacteria effect (i.e. the number of raphidophyte species affected by a particular bacteria strain) and raphidophyte susceptibility (i.e. the number bacteria strains affecting a particular raphidophyte species), a toxic effect was always found when strains of a raphidophyte species were exposed to algicidal bacteria isolated from a bloom caused by that same species. The results suggest that algicidal bacteria may be an important limiting factor in raphidophyte bloom sustenance and can promote bloom decline in brackish lagoonal eutrophic estuaries.     

西藏拉萨地区常见致病菌及其耐药性分析

目的研究2009年至2011年西藏拉萨地区主要医院的常见致病菌及其耐药性情况。方法采集拉萨市临床医院细菌感染性疾病临床标本1 200份进行致病菌的分离。采用法国梅里埃-ATB菌种鉴定仪对分离的菌株鉴定到种,参照2010年CLSI推荐方法进行耐药性分析。结果对拉萨地区主要临床医院的感染者标本分离鉴定出332株临床致病菌,其中细菌304株(91.57%),真菌28株(8.43%)。病原细菌分布主要为革兰阳性球菌97株,占29.22%;革兰阴性杆菌200株,占60.24%;其他菌7株,占2.11%。凝固酶阴性葡萄球菌对苯唑西林﹑头孢曲松和环丙沙星的耐药率分别为72%﹑40%和44%。大肠埃希菌对氨苄西林﹑哌拉西林和头孢唑林耐药率分别为83%﹑53%和43%。克雷伯菌属对氨苄西林﹑头孢唑林耐药率分别为86%和58%。铜绿假单胞菌和不动杆菌对亚胺培南的耐药率分别高达20%和19%。结论拉萨地区的细菌感染及其耐药菌株分布较为广泛,该地区应加强常规临床致病菌的耐药性监测以指导临床医师合理使用抗菌药物。     

The antimicrobial activity of Aspergillus fumigatus is enhanced by a pool of bacteria

In a screening program for new antibiotic producers, a strain of Aspergillus fumigatus was isolated from Brazilian soil samples. A pool of autoclaved bacteria was added to part of the fungus culture on the second day of fermentation to increase antibiotic production. The chloroform extract from the culture broth to which the pool of autoclaved bacteria was added showed an increase of 55%, 63% and more than 100% in activity against Staphylococcus aureus, Candida albicans and Micrococcus luteus, respectively. Also, the HPLC chromatographic profiles of the chloroform extracts from both culture conditions were different. Two active compounds were isolated from the broth of the culture grown in the presence of pooled bacteria and were identified as 3,4-dimethoxyphenol and 1,3,5-trimethoxybenzene.     

High efficiency transduction of single strand plasmid DNA into enteric bacteria

Summary This report demonstrates high efficiency transduction of enteric bacteria using single strand plasmids packaged in M13 phage capsids. Transformation by plasmid DNA is usually a very inefficient process in many enteric bacteria other than Escherichia coli K12. Plasmids carrying an M13 origin of replication can be replicated and packaged when cells carrying such plasmids are infected with M13 or a derivative helper phage. By introducing an F plasmid into E. coli, Serratia marcescens, Citrobacter freundii, and Enterobacter aerogenes, these species can now be infected at high efficiency with M13 phage and with packaged single strand plasmids, yielding an efficient method to introduce cloned DNA fragments into these bacteria. The titer of colony forming units in a lysate was essentially equivalent in all the bacteria, demonstrating an equal efficiency of transduction of these other enteric bacteria compared to E. coli.