High colonization by native arbuscular mycorrhizal fungi (AMF) of rubber trees in small-holder plantations on low fertility soils in North East Thailand

Rubber tree is a very important crop in Thailand, representing an essential source of income for farmers. In the past two decades, rubber tree plantations have been greatly expanding in unfavorable areas, where climate conditions are difficult and soil fertility is very poor. To optimize latex yields, mineral fertilizers have been widely used. A better understanding of the roles of the biological compartment in soil fertility is essential to determine alternative management practices to sustain soil fertility and optimize latex yields. Arbuscular mycorrhizal fungi (AMF) are widely recognized as beneficial for plants, mainly through their role in improving plant nutrient uptake. The objective of this study was to assess the AMF populations in rubber tree plantations and the impact of both soil characteristics and plantation age on these communities. Our results showed that all rubber trees were highly colonized, regardless of the soil structure and nutrient contents. AMF colonization was not affected by the age of the trees, suggesting that maintaining the symbiosis is likely to be beneficial at all stages. A better understanding and management of the microbial communities would contribute to maintaining or restoring soil fertility, leading to a better tree growth and optimized latex yield.

Diversity of root-associated arbuscular mycorrhizal fungal communities in a rubber tree plantation chronosequence in Northeast Thailand

Rubber tree (Hevea brasiliensis) is of major economic importance in Southeast Asia and for small land holders in Thailand in particular. Due to the high value of latex, plantations are expanding into unsuitable areas, such as the northeast province of Thailand where soil fertility is very low and therefore appropriate management practices are of primary importance. Arbuscular mycorrhizal fungi (AMF) contribute to plant growth through a range of mechanisms and could play a key role in a more sustainable management of the rubber plantations. We described the diversity of AMF associated with rubber tree roots in Northeast Thailand in relation to tree age and soil parameters along a chronosequence of rubber tree plantations. Cassava fields were included for comparison. Rubber tree and cassava roots harbored high diversity of AMF (111 Virtual Taxa, VT), including 20 novel VT. AMF VT richness per sample was consistently high (per site mean 16 to 21 VT per sample) along the chronosequence and was not related to soil properties. The composition of AMF communities differed between cassava and rubber tree plantations and was influenced by soil texture and nutrient content (sand, K, P, Ca). AMF community composition gradually shifted with the age of the trees. Our results suggest that the high diversity of AMF in this region is potentially significant for maintaining high functionality of AMF communities.

Rhizophagus irregularis as an elicitor of rosmarinic acid and antioxidant production by transformed roots of Ocimum basilicum in an in vitro co-culture system

Arbuscular mycorrhiza is a symbiotic association formed between plant roots and soil borne fungi that alter and at times improve the production of secondary metabolites. Detailed information is available on mycorrhizal development and its influence on plants grown under various edapho-climatic conditions, however, very little is known about their influence on transformed roots that are rich reserves of secondary metabolites. This raises the question of how mycorrhizal colonization progresses in transformed roots grown in vitro and whether the mycorrhizal fungus presence influences the production of secondary metabolites. To fully understand mycorrhizal ontogenesis and its effect on root morphology, root biomass, total phenolics, rosmarinic acid, caffeic acid and antioxidant production under in vitro conditions, a co-culture was developed between three Agrobacterium rhizogenes-derived, elite-transformed root lines of Ocimum basilicum and Rhizophagus irregularis. We found that mycorrhizal ontogenesis in transformed roots was similar to mycorrhizal roots obtained from an in planta system. Mycorrhizal establishment was also found to be transformed root line-specific. Colonization of transformed roots increased the concentration of rosmarinic acid, caffeic acid and antioxidant production while no effect was observed on root morphological traits and biomass. Enhancement of total phenolics and rosmarinic acid in the three mycorrhizal transformed root lines was found to be transformed root line-specific and age dependent. We reveal the potential of R. irregularis as a biotic elicitor in vitro and propose its incorporation into commercial in vitro secondary metabolite production via transformed roots.

Mycorrhizal elicitation of secondary metabolites in hairy roots of Ocimum basilicum

 Mycorrhiza, a symbiotic soil fungus was identified as a biotic elicitor of antioxidant compounds found in the plant roots. In vitro developed technique and bioresources carry potential towards formation of biological and biochemical factories for application in the agricultural and pharmaceutical industries.

Spatial model for predicting the presence of cinnamon fungus (Phytophthora cinnamomi) in sclerophyll vegetation communities in south-eastern Australia

The pathogen Phytophthora cinnamomi causes extensive 'dieback' of Australian native vegetation. This study investigated the distribution of infection in an area of significant sclerophyll vegetation in Australia. It aimed to determine the relationship of infection to site variables and to develop a predictive model of infection. Site variables recorded at 50 study sites included aspect, slope, altitude, proximity to road and road characteristics, soil profile characteristics and vegetation attributes. Soil and plant tissues were assayed for the presence of the pathogen. A geographical information systyem (GIS) was employed to provide accurate estimations of spatial variables and develop a predictive model for the distribution of P. cinnamomi. The pathogen was isolated from 76% of the study sites. Of the 17 site variables initially investigated during the study a logistic regression model identified only two, elevation and sun-index, as significant in determining the probability of infection. The presence of P. cinnamomi infection was negatively associated with elevation and positively associated with sun-index. The model predicted that up to 74% of the study area (11 875 ha) had a high probability of being affected by P. cinnamomi. However, the present areas of infection were small, providing an opportunity for management to minimize spread into highly susceptible uninvaded areas.

Southern Boobook Ninox novaeseelandiae feeding on the Fungus Pislithus albus in North Queensland

This note describes an observation of a Southern Boobook Ninox ovaeseelandiae pecking at the fruiting body of a terrestrial fungus in northern Queensland.

Identification and possible disease mechanisms of an under-recognized fungus, Aureobasidium pullulans

Background: Investigations into the occurrence and health effects of yeast-like fungi in the outdoor air in the US have been limited. We sought to identify a respirable-sized fungus common in the Pasadena air, locate a major source for the emissions and investigate its relevance to allergic disease. Methods: Yeast-like fungi sampled from the environment were isolated, microscopically examined and sequenced. Pasadena allergy patients were skin tested with commercially available fungal extracts. Patient serum was immunoanalyzed for specific IgE reactivity. Nearby vegetation was analyzed in a controlled emission chamber to find a major source for the aerosols. Results: Hyaline unicellular conidia comprised up to 90% (41,250 m^-3 of air) of total fungal counts and generally peaked at night and during periods of rainfall and ensuing winds throughout the fall and winter. Flowers were determined to be a major source of the emissions. The cellular and colonial morphology of isolates were consistent with Aureobasidium species. The sequence of the D1/D2 region of the 26S ribosomal subunit of isolates from flowers showed identity to two strains of Aureohasidium pullulans (black yeast). Seventeen percent (16/94) of atopic individuals had positive skin testing with A. pullulans extract. Patient sera gE identified several high molecular weight allergens in Aureobasidium extracts. Conclusions: Respirable-sized conidia of A. pullulans are emitted from flowers and form high concentrations in the air. They are associated with immediate reactivity on skin tests, bind to patient sera IgE, and might be relevant in allergic upper and lower airway diseases.

Landscape context influences chytrid fungus distribution in an endangered European amphibian

Wildlife disease is an emerging threat to biodiversity. The amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd), which causes the disease chytridiomycosis, has been documented in over 500 amphibian species globally. Understanding conditions under which amphibians are vulnerable to Bd is important for evaluating species risk and developing surveillance strategies. Here, we investigate the spatial distribution of Bd infection in the ephemeral pond-breeding yellow-bellied toad Bombina variegata, a species of high conservation concern in the European Union. We sampled 550 toads from 60 ponds in a traditional agricultural landscape in Southern Transylvania, Romania. Overall, Bd prevalence was low in B.variegata, but infected toads were widely dispersed through the landscape and were found in a quarter of all sampled ephemeral ponds. At the pond level, increased Bd occurrence was associated with short distances to perennial water sources and high forest cover. These findings suggest that perennial water sources may act as source habitat for Bd, with amphibian movements resulting in Bd spillover into ephemeral ponds. Increased Bd occurrence in ponds surrounded by high levels of forest cover is likely related to cooler and wetter conditions that are more favourable for Bd. Throughout the study landscape, patchy environmental suitability for Bd appears to restrict the pathogen to a subset of B.variegata habitat. Ephemeral ponds in open landscapes, without nearby perennial habitat, likely provide an environmental refuge from Bd, where the risk of infection is low. From a conservation perspective, these findings highlight the importance of maintaining ephemeral ponds in open landscapes, but these are currently threatened by land-use change.

Decline and re-expansion of an amphibian with high prevalence of chytrid fungus

The disease chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), is a key driver of global amphibian declines. While chytridiomycosis can cause extinction, many susceptible species persist after an initial period of decline, albeit with reduced abundance and distribution. Emerging evidence indicates that amphibian abundance can recover within remnant populations, but to date, the capacity of amphibian populations to re-expand into historically occupied habitat has received limited research attention. We surveyed 145 sites in 2011 and 2012 to determine if populations of the whistling tree frog (Litoria verreauxii verreauxii) have re-expanded compared with historical data from 1975-1976, 1990 and 1996. L. v. verreauxii underwent a major range contraction likely caused by chytridiomycosis between the first two time periods. Populations have recently re-expanded, with 39 new sites colonised despite high prevalence of Bd. We suspect that changes in disease dynamics have resulted in the increased coexistence of L. v. verreauxii and Bd. Habitat attributes at sites that retained frogs for the duration of the study indicate that high quality habitat may contribute to buffering against population level effects of Bd. Colonised sites had more coarse woody debris, suggesting a possible habitat management strategy to encourage range expansion for this species. Given sufficient time and adequate source populations in high quality habitat, it is possible that other amphibian species may re-expand from chytridiomycosis-induced declines. This provides an impetus for the protection of historical, but currently unoccupied amphibian habitats and highlights the importance of maintaining high quality habitat to help species survive novel shocks such as pandemic diseases.

Degradation mechanisms of benzo[a]pyrene and its accumulated metabolites by biodegradation combined with chemical oxidation

A high degradation extent of benzo[a]pyrene (BaP) should not be considered as the sole desirable criterion for the bioremediation of BaP-contaminated soils because some of its accumulated metabolites still have severe health risks to human. Two main metabolites of BaP, benzo[a]pyrene-1,6-quinone (BP1,6-quinone) and 3-hydroxybenzo[a]pyrene (3-OHBP) were identified by high performance liquid chromatography (HPLC) with standards. This study was the first time that degradation of both BaP and the two metabolites was carried out by chemical oxidation and biodegradation. Three main phases during the whole degradation process were proposed.

Hydrogen peroxide–zinc (H₂O₂–Zn), the fungus – Aspergillus niger and the bacteria – Zoogloea sp. played an important role in the different phases. The degradation parameters of the system were also optimized, and the results showed that the effect of degradation was the best when fungus–bacteria combined with H₂O₂–Zn, the concentration range of BaP in the cultures was 30–120 mg/l, the initial pH of the cultures was 6.0. However, as co-metabolites, phenanthrene significant inhibited the degradation of BaP. This combined degradation system compared with the conventional method of degradation by domestic fungus only, enhanced the degradation extent of BaP by more than 20% on the 12 d. The highest accumulation of BP1,6-quinone and 3-OHBP were reduced by nearly 10% in the degradation experiments, which further proved that the combined degradation system was more effective as far as joint toxicity of BaP and its metabolites are concerned.

Field-scale bioremediation of soil contaminated with crude oil

Field-scale remediation of oil-contaminated soils from the Liaohe Oil Fields in China was examined using composting biopiles in windrow technology. Micronutrient-enriched chicken excrement and rice husk were applied as nutrition and a bulking agent. The lipase activities of indigenous micro-organisms were analyzed, and three indigenous fungi with high lipase activities was identified. An inoculum consisting of the three indigenous fungi and one introduced (exotic) fungus was applied to four different types of oil-contaminated soils. The results showed that the inoculum of indigenous fungi increased both the total colony-forming units (TCFU) and increased the rate of degradation of total petroleum hydrocarbons (TPH) in all contaminated soils but at different rates. In sharp contrast to other studies, the introduction of exotic micro-organisms did not improve the remediation, and suggests that inoculation of oil-contaminated sites with nonindigenous species is likely to fail. On the other hand, indigenous genera of microbes were found to be very effective in increasing the rate of degradation of TPH. The degradation of TPH was mainly controlled by the compositions of aromatic hydrocarbons and asphaltene and resin. Between 38 to 57% degradation of crude oils (with densities ranging from 25,800 to 77,200 mg/kg dry weight) in contaminated soils was achieved after 53 days of operation. The degradation patterns followed typical first-order reactions. We demonstrate that the construction and operation of field-scale composting biopiles in windrows with passive aeration is a cost-effective bioremediation technology.

Effect of Phytophthora cinnamomi on the habitat utilization of Antechinus stuartii in a Victorian forest

Phytophthora cinnamomi (Cinnamon fungus) is a pathogenic soil fungus which infects plant communities along the south-eastern coast of Australia, and the south-western corner of Western Australia. The symptoms of this disease include chlorosis, death of branches (ie. ‘dieback’), retarded growth and the eventual death of infected plants. This leads to devastating effects upon plant communities by altering both the structural and floristic characteristics of these communities. Small mammal species are dependent on specific features of their habitat such as vegetation structure and floristics. This thesis investigated alterations to the habitat of the insectivorous marsupial mouse, Antechinus stuartii, due to the presence of P. cinnamomi. The study was undertaken in an area of an open forest in the Brisbane Ranges, Victoria. Significant changes were found in both the floristic composition and structure of the vegetation at study sites infected with P, cinnamomi, compared to uninfected sites. The habitat utilization by A. stuartii of uninfected and infected vegetation was investigated using live trapping and radio-telemetric techniques. Capture rates were higher at sites uninfected by P. cinnamomi, and both male and females selected areas free from infection. Home range areas of males were significantly larger than those of females as assessed by telemetry. Both sexes spent a high proportion of time in areas dominated by Xanthorrhoea australis (Austral grass tree). There were significant relationships between the abundance of A. stuartii and the denseness of vegetation above 1 metre in height, and in particular, the proportion of cover afforded by X. australis. There were no significant differences in the cover of Eucalyptus spp. between uninfected and infected sites, but there were significantly more nest hollows in infected areas. The abundance of invertebrates was examined using pitfall traps. There were no significant differences in the abundance of the larger invertebrate taxa at infected and uninfected sites, but higher abundances of some micro-invertebrate groups in infected areas were recorded. The most likely factors considered to be influential in the habitat selection of A. stuartii were vegetation structure, and the presence of X. australis. To assess whether these factors were important the leaves of X. australis were removed with a brushcutter, to mimic the early effects of infection with P. cinnamomi. Animals did not respond to the alteration of vegetation structure in the short term (3-4 days). Longer-term experiments are required to assess the habitat utilization of A. stuartii at different periods following habitat manipulation. The implications of the presence of P. cinnamomi on the conservation of fauna are discussed. The destructive nature of the pathogen, and the slow rate of recovery from the disease, means that P. cinnamomi can be considered a threatening process to plant communities and the fauna that reside within that habitat. Future management of this disease within natural areas must therefore be cognisant of the potential of P. cinnamomi to significantly affect faunal as well as vegetative communities.

Self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin EAS

The hydrophobin EAS from the fungus Neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. Self-assembly of EAS into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. We have used site-directed mutagenesis and peptide experiments to identify the region of EAS that drives intermolecular association and formation of the cross-β rodlet structure. Transplanting this region into a nonamyloidogenic hydrophobin enables it to form rodlets. We have also determined the structure and dynamics of an EAS variant with reduced rodlet-forming ability. Taken together, these data allow us to pinpoint the conformational changes that take place when hydrophobins self-assemble at an interface and to propose a model for the amphipathic EAS rodlet structure.

Fungal Mastoiditis in immunocompromised children

The immunocompromised host is subject to a variety of opportunistic infections. Mycotic infections, including invasive fungal sinusitis, are a dreaded complication in immune deficient children. Fungal mastoiditis has rarely been described in this population. Our experience with 2 cases of fungal mastoiditis in immunocompromised children is reviewed. Case histories describing aggressive medical management with and without surgical intervention and a review of the literature are presented.Fungal mastoiditis is a rare entity described in isolated case reports in the adult literature. It is seen almost entirely in immunocompromised patients, particularly those lacking cell-mediated immunity. The first case of Aspergillus mastoiditis was described in 1985.1 Reports of fungal mastoiditis have been primarily of patients with leukemia, and, more recently, of patients with acquired immunodeficiency syndrome.2,3 Using a computerized search of the MEDLINE database, we identified 1 report (in a non–English language journal) of fungal mastoiditis in a pediatric patient.4 We report 2 cases of fungal mastoiditis in immunosuppressed children.