Antibacterial activities of naturally occurring compounds against antibiotic-resistant Bacillus cereus vegetative cells and spores, Escherichia coli, and Staphylococcus aureus

After demonstrating the lack of effectiveness of standard antibiotics against the acquired antibiotic resistance of Bacillus cereus (NCTC 10989), Escherichia coli (NCTC 1186), and Staphylococcus aureus (ATCC 12715), we showed that the following natural substances were antibacterial against these resistant pathogens: cinnamon oil, oregano oil, thyme oil, carvacrol, (S)-perillaldehyde, 3,4-dihydroxybenzoic acid (beta-resorcylic acid), and 3,4-dihydroxyphenethylamine (dopamine). Exposure of the three pathogens to a dilution series of the test compounds showed that oregano oil was the most active substance. The oils and pure compounds exhibited exceptional activity against B. cereus vegetative cells, with oregano oil being active at nanogram, per milliliter levels. In contrast, activities against B. cereus spores were very low. Activities of the test compounds were in the following approximate order: oregano oil > thyme oil approximate to carvacrol > cinnamon oil > perillaldehyde > dopamine > beta-resorcylic acid. The order of susceptibilities of the pathogens to inactivation was as follows: B. cereus (vegetative) much greater than S. aureus approximate to E. coli much greater than B. cereus (spores). Some of the test substances may be effective against antibiotic-resistant bacteria in foods and feeds.

Factors influencing the activity of taurolidine against spores of Bacillus subtilis (NCTC 10073)

The cidal activities of aqueous taurolidine (2.0% w/v containing 5.0% wlv polyvinylpyrrolidone as a solubilising agent) and alcoholic taurolidine (2.0% w/v dissolved in Isopropyl alcohol 50% v/v) against spores of Bacillus subtilis NCTC 10073 were evaluated at 20 degrees C, 37 degrees C, 45 degrees C and 55 degrees C. Increased temperature increased both the rate and extent of sporicidal activity of both solutions. Total spore kill was not observed in either solution type over the range of temperatures and contact times examined. There were no observed differences between the sporicidal activities of aqueous and alcoholic taurolidine solutions at all temperatures examined. Ultrasonic energy (50 Hz operating frequency in a 150 W ultrasonic bath in conjunction with increasing temperature allowed to rise naturally from ambient temperature to 41 degrees C over 4 h) enhanced the sporicidal activities of both solution types. However, the difference in activity between the two solution types was not significant. Compared to normal spores, alteration of spore coat layers (hydrogen-form spores) did not alter spore susceptibility to aqueous taurolidine at elevated temperatures of 37 degrees C and 55 degrees C.

Manipulation of intracellular glycerol and erythritol enhances germination of conidia at low water availability

The insect pathogen Beauveria bassiana, Metarhizium anisopliae and Paecilomyces farinosos can be effective biocontrol agents when relative humidity (RH) is close to 100%. At reduced water availability, germination of propagules, and therefore host infection, cannot occur. Cultures of B. bassiana, M. anisopliae and P. farinosus were grown under different conditions to obtain conidia with a modified polyol and trehalose content. Conidia with higher intracellular concentrations of glycerol and erythritol germinated both more quickly and at lower water activity (a(w)) than those from other treatments. In contrast, conidia containing up to 235.7 mg trehalose g-1 germinated significantly (P < 0 05) more slowly than those with an equivalent polyol content but less trehalose, regardless of water availability. Conidia from control treatments did not germinate below 0.951 - 0.935 a(w) (≡ 95.1 - 93.5% RH). In contrast, conidia containing up to 164.6 mg glycerol plus erythritol g-1 germinated down to 0.887 a(w) (≡ 88.7% RH). These conidia germinated below the water availability at which mycelial growth ceases (0.930 - 0.920 a(w)). Germ tube extension rates reflected the percentage germination of conidia, so the most rapid germ tube growth occurred after treatments which produced conidia containing the most glycerol and erythritol. This study shows for the first time that manipulating polyol content can extend the range of water availability over which fungal propagules can germinate. Physiological manipulation of conidia may improve biological control of insect pests in the field.

Arbuscular mycorrhizal fungi confer enhanced arsenate resistance on Holcus lanatus

The role of arbuscular mycorrhizal fungi (AMF) in arsenate resistance in arbuscular mycorrhizal associations is investigated here for two Glomus spp. isolated from the arsenate-resistant grass Holcus lanatus. Glomus mosseae and Glomus caledonium were isolated from H. lanatus growing on an arsenic-contaminated mine-spoil soil. The arsenate resistance of spores was compared with nonmine isolates using a germination assay. Short-term arsenate influx into roots and long-term plant accumulation of arsenic by plants were also investigated in uninfected arsenate resistant and nonresistant plants and in plants infected with mine and nonmine AMF. Mine AMF isolates were arsenate resistant compared with nonmine isolates. Resistant and nonresistant G. mosseae both suppressed high-affinity arsenate/phosphate transport into the roots of both resistant and nonresistant H. lanatus. Resistant AMF colonization of resistant H. lanatus growing in contaminated mine spoil reduced arsenate uptake by the host. We conclude that AMF have evolved arsenate resistance, and conferred enhanced resistance on H. lanatus.

Relative toxicity of arsenite and arsenate on germination and early seedling growth of rice (Oryza sativa L.)

Elevated soil arsenic levels resulting from long-term use of arsenic contaminated ground for irrigation in Bangladesh may inhibit seed germination and seedling establishment of rice, the country's main food crop. A germination study on rice seeds and a short-term toxicity experiment with different concentrations of arsenite and arsenate on rice seedlings were conducted. Percent germination over control decreased significantly with increasing concentrations of arsenite and arsenate. Arsenite was found to be more toxic than arsenate for rice seed germination. There were varietal differences among the test varieties in response to arsenite and arsenate exposure. The performance of the dry season variety Purbachi was the best among the varieties. Germination of Purbachi was not inhibited at all up to 4 mg l^-1 arsenite and 8 mg l^-1 arsenate treatment. Root tolerance index (RTI) and relative shoot height (RSH) for rice seedlings decreased with increasing concentrations of arsenite and arsenate. Reduction of RTI caused by arsenate was higher than that of arsenite. In general, dry season varieties have more tolerance to arsenite or arsenate than the wet season varieties.

Hygrothermal performance of wood-hemp insulation in timber frame wall panels with and without a vapour barrier

An in situ experiment on a full-scale timber frame test building was carried out to study the hygrothermal performance of wood-hemp composite insulation in timber frame wall panels with and without a vapour barrier. The heat transfer properties and the likelihood of mould growth and condensation in the panels were compared. Step changes in the internal relative humidity were performed to explore the effects of high, normal and low internal moisture loads on the wall panels. No significant difference in the average equivalent thermal transmittance (U-values) between the panels with and without a vapour barrier was observed. The average equivalent U-values of the panels were close to the U-values calculated from the manufacturers’ declared thermal conductivity values of the insulation. The likelihood of condensation was higher at the interface of the wood-hemp insulation and the oriented strand board (OSB) in the panel without a vapour barrier. In terms of the parametric assessment of the mould germination potential, the relative humidity, the temperature and the exposure conditions in the insulation-OSB interfaces of the panel without a vapour barrier were found to be more favourable to the germination of mould spores. Nonetheless, when the insulations were dismantled, no mould was visually detected.

Germination requirements of Plantago algarbiensis seeds, an endangered species endemic to the Algarve region (South of Portugal)

The ecdysteroid, 20-hydroxyecdysone or beta-ecdysone, is a steroid hormone which plays a crucial role in molting, metamor- phosis and reproduction of arthropods. This ecdysteroid and its analogues have high potential to be used as insecticides. Previous studies in our laboratory have demonstrated that Vitex glabrata R.Br. (commonly known as Khai-Nao), an indigenous herbaceous plant of Thailand, synthesized and accumulated high quantity of 20-hydroxyecdysone. Therefore, the aim of this study was to investigate the effect of precursor and elicitors feeding on cell growth and 20-hydroxyecdysone production of V. glabrata suspension cultures. Plant cells were cultured in half strength MS medium containing 30 g/l glucose and supplemented with 2.0 mg/l 6- benzylaminopurine (BAP) and 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Cells were incubated on a rotary shaker at 120 rpm under continuous light of 2000 lux at 25 °C. Sterilized cholesterol (5 and 10 mg/l) as precursor was added to the cell cultures on the day of inoculation, while chitosan (50, 100 and 200 mg/l) and methyl jusmonate (100 and 200 mM) as elicitors were added to the cell cultures on day 6 after cultivation.

Back-trajectories Show Export of Airborne Fungal Spores (Ganoderma sp.) From Forests to Agricultural and Urban Areas in England

We propose here the hypothesis that all of United Kingdom (UK) is likely to be affected by Ganoderma sp. spores, an important plant pathogen. We suggest that the main sources of this pathogen, which acts as a bioaerosol, are the widely scattered woodlands in the country, although remote sources must not be neglected. The hypothesis is based on related studies on bioaerosols and supported by new observations from a non-forest site and model calculations to support our hypothesis. Hourly concentrations of Ganoderma sp. spores were measured from 2006 to 2010 using a 7-day volumetric spore trap at the city of Worcester. The concentrations peak during the night and early in the morning. This suggests that the main spore sources are located a few hours away with respect to air masses transport and reach urban areas thanks to air masses transport. The back-trajectory analysis was applied to determine the location of Ganoderma sp. spore sources. The analysis of back-trajectories demonstrated that 78% of the air masses reached Worcester from a 180° arc direction from the East to West. Three episodes were selected for detailed investigation and they revealed that during the episodes air masses always passed main UK woodlands before the arrival in Worcester, independently of their origin, but the long distance transport under certain conditions might be possible. Our studies suggest that the sources of UK Ganoderma sp. spores are mainly to be found in UK. Hence our studies suggest that research and mitigation strategies in UK should give their main attention to national sources, without neglecting the contribution from long distance transport.

Atmospheric Concentrations of Alternaria, Cladosporium, Ganoderma and Didymella Spores Monitored in Cork (Ireland) and Worcester (England) During the Summer of 2010

This study represents the first international intercomparison of fungal spore observations since 1990, focusing on atmospheric concentrations of Alternaria, Cladosporium, Ganoderma and Didymella spores. The campaigns were performed at sites located in Cork (Ireland) and Worcester (England) during summer 2010. Observations were made using Hirst-type volumetric spore traps and corresponding optical identification at the genus level by microscope. The measurements at both sites (including meteorological parameters) were compared and contrasted. The relationships between the fungal spore concentrations with selected meteorological parameters were investigated using statistical methods and multivariate regression trees (MRT). The results showed high correlations between the two sites with respect to daily variations. Statistically significant higher spore concentrations for Alternaria, Cladosporium and Ganoderma were monitored at the Worcester site. This result was most likely due to the differences in precipitation and local fungal spore sources at the two sites. Alternaria and Cladosporium reached their maxima a month earlier in Cork than in Worcester, and Didymella with Ganoderma peaked simultaneously with similar diurnal trends found for all the investigated spore types. MRT analysis helped to determine threshold values of the meteorological parameters that exerted most influence on the presence of spores: they were found to vary at the two sites. Our results suggest that the aeromycological profile is quite uniform over the British Isles, but a description of bioaerosols with respect to overall load and daily concentration can be quite diverse although the geographical difference between sites is relatively small. These variations in the concentrations therefore need to be explored at the national level

Potential Sources of Airborne Alternaria spp. spores in South-west Spain

Fungi belonging to the genus of Alternaria are recognised as being significant plant pathogens, and Alternaria allergens are one of themost important causes of respiratory allergic diseases in Europe. This study aims to provide a detailed and original analysis of Alternaria transport dynamics in Badajoz, SW Spain. This was achieved by examining daily mean and hourly observations of airborne Alternaria spores recorded during days with high airborne concentrations of Alternaria spores (N100 s m−3) from 2009 to 2011, as well as four inventory maps of major Alternaria habitats, the overall synoptic weather situation and analysis of air mass transport using Hybrid Single Particle Lagrangian Integrated Trajectory model and geographic information systems. Land use calculated within a radius of 100 km from Badajoz shows that crops and grasslands are potentially the most important local sources of airborne Alternaria spores recorded at the site. The results of back trajectory analysis showthat, during the examined four episodes, the two main directions where Alternaria source areas were located were: (1) SW–W; and (2) NW–NE. Regional scale and long distance transport could therefore supplement the airborne catch recorded at Badajoz with Alternaria conidia originating from sources such as crops and orchards situated in other parts of the Iberian Peninsula.

Investigating and Predicting Atmospheric Concentrations of Allergenic Fungal Spores (Alternaria, Cladosporium, Didymella and Ganoderma)

Air quality is an increasing concern of the European Union, local authorities, scientists and most of all inhabitants that become more aware of the quality of the surrounding environment. Bioaerosols may be consisted of various elements, and the most important are pollen grains, fungal spores, bacteria, viruses. More than 100 genera of fungal spores have been identified as potential allergens that cause immunological response in susceptible individuals. Alternaria and Cladosporium have been recognised as the most important fungal species responsible for respiratory tract diseases, such as asthma, eczema, rhinitis and chronic sinusitis. While a lot of attention has been given to these fungal species, a limited number of studies can be found on Didymella and Ganoderma, although their allergenic properties were proved clinically. Monitoring of allergenic fungal spore concentration in the air is therefore very important, and in particular at densely populated areas like Worcester, UK. In this thesis a five year spore data set was presented, which was collected using a 7-day volumetric spore trap, analysed with the aid of light microscopy, statistical tests and geographic information system techniques. Although Kruskal-Wallis test detected statistically significant differences between annual concentrations of all examined fungal spore types, specific patterns in their distribution were also found. Alternaria spores were present in the air between mid-May/mid-June until September-October with peak occurring in August. Cladosporium sporulated between mid-May and October, with maximum concentration recorded in July. Didymella spores were seen from June/July up to September, while peaks were found in August. Ganoderma produced spores for 6 months (May-October), and maximum concentration could be found in September. With respect to diurnal fluctuations, Alternaria peaked between 22:00h and 23:00h, Cladosporium 13:00-15:00h, Didymella 04:00-05:00h and 22:00h-23:00h and Ganoderma from 03:00h to 06:00h. Spatial analysis showed that sources of all fungal species were located in England, and there was no evidence for a long distance transport from the continent. The maximum concentration of spores was found several hours delayed in comparison to the approximate time of the spore release from the crops. This was in agreement with diurnal profiles of the spore concentration recorded in Worcester, UK. Spores of Alternaria, Didymella and Ganoderma revealed a regional origin, in contrast to Cladosporium, which sources were situated locally. Hence, the weather conditions registered locally did not exhibit strong statistically significant correlations with fungal spore concentrations. This has had also an impact on the performance of the forecasting models. The best model was obtained for Cladosporium with 66% of the accuracy.