Candida keroseneae sp. nov., a novel contaminant of aviation kerosene.

AIMS: To characterize and identify a novel contaminant of aviation fuel. METHODS AND RESULTS: Micro-organisms (yeasts and bacteria) were isolated from samples of aviation fuel. A yeast that proved to have been unrecorded previously was isolated from more than one fuel sample. This novel yeast proved to be a new species of Candida and is described here. Ribosomal RNA gene sequence analyses of internal transcribed spacer (ITS) regions (including 5·8S subunit) plus the 26S D1/D2 domains showed the strains to cluster within the Candida membranifaciens clade nearest to, but distinct from, Candida tumulicola. Phenotypic tests were identical for both isolates. Physiological and biochemical tests supported their position as a separate taxon. The yeast was assessed for its effect on the main constituent hydrocarbons of aviation fuel. CONCLUSIONS: Two strains (IMI 395605(T) and IMI 395606) belonging to the novel yeast species, Candida keroseneae, were isolated from samples of aircraft fuel (kerosene), characterized and described herein with reference to their potential as contaminants of aviation fuel. SIGNIFICANCE AND IMPACT OF THE STUDY: As a result of isolating a novel yeast from aviation fuel, the implications for microbial contamination of such fuel should be considered more widely than previously thought.

Measurement and repeatability of interrupter resistance in unsedated newborn infants.

Interrupter resistance (R(int)) is a useful measure of airway caliber in young children, but has not been well characterized in infants-in whom there are concerns about the accurate measurement of driving pressure. This study aimed to assess the feasibility and repeatability of measuring R(int) in unsedated newborn infants, and to explore alternative algorithms for calculating driving pressure. R(int) measurement was attempted in 28 healthy term newborn infants during natural sleep using the MicroRint device. Paired R(int) measurements were achieved in 24 infants, but after screening of waveforms only 15 infants had at least 5 technically acceptable waveforms on both measurements. R(int) values obtained were comparable with reported values for airflow resistance in newborns using other methods. However, the repeatability coefficient (CR) was much higher than reported values in preschool children using standard back-extrapolation algorithms, with CR 2.47 KPa L(-1) sec (unscreened) and 2.93 KPa L(-1) sec (screened). Other algorithms gave only marginally better repeatability, with all CR values over 50% of the mean R(int) value. Using current commercially available equipment, R(int) is too poorly repeatable to be a reliable measurement of airflow resistance in newborn infants. Lower deadspace equipment is needed, but anatomical and physiological factors in the infant are also important.

An association between the Antarctic mite Alaskozetes antarcticus and an entomophthoralean fungus of the genus Neozygites.

A fungal pathogen provisionally identified as Neozygites cf. acaridis has recently been isolated from the Antarctic oribatid mite Alaskozetes antarcticus. The identification of the fungus is discussed with reference to recent changes in the taxonomy of Neozygites. The potential role of the fungus in the Antarctic mite populations is considered in relation to the known mite life cycles, and the particular environmental conditions in the Antarctic.

Domain II hairpin structure in ITS1 sequences as an aid in differentiating recently evolved animal and plant pathogenic fungi.

The hypothesis that ITS structural features can be used to define fungal groups, where sequence analysis is unsatisfactory, was examined in plant and animal pathogenic fungi. Structural models of ITS1 regions were predicted for presumed closely related species in Colletotrichum and Trichophyton anamorphs of Arthroderma species. Structural alignment of models and comparison with ITS sequence analysis identified a variable region in a conserved hairpin formed from a common inverted repeat. Thirteen different hairpin structure models were obtained for Colletotrichum species and five different models were obtained for Trichophyton species. The different structure types could be matched to individual species and species complexes as defined by ITS sequence analysis.

Lung function measurement in prematurely born preschool children with and without chronic lung disease.

OBJECTIVE: Prematurely born infants often have recurrent wheeze and long-term respiratory morbidity at follow-up. Assessment of airways obstruction in preschool children is feasible using the interrupter resistance (Rint) but has rarely been examined in preterm children with and without chronic lung disease (CLD). The objective of this study was to determine lung function measured by the interrupter technique, its feasibility in the ambulatory setting and respiratory health in prematurely born preschool children with and without CLD. STUDY DESIGN: Preterm children of 2 to 4 years with severe CLD (>30% oxygen at 36 weeks and discharged home receiving supplemental oxygen) (n=43, median gestational age 27 weeks and median birth weight 995 g) and without CLD (n=33, median gestational age 29 weeks and median birth weight 1366 g) attempting lung function test for the first time were enrolled. Respiratory symptoms score was calculated using a questionnaire. A single set of 10 consecutive Rint measurements was obtained using a portable device (MicroRint). Median of at least five occlusions with consistent shape of mouth pressure-time curves was taken to be a Rint measurement. To assess feasibility the children were categorized as 'satisfactory', 'failure' and 'rejected' depending on the outcome of the test. Outcome variables were respiratory symptoms score and Rint. RESULT: Satisfactory Rint measurement was obtained in 46 (61%) children, 9 (36%) 2-year olds, 17 (65%) 3-year olds and 20 (80%) 4-year olds. As compared with the preterm control children (n=18), CLD children (n=28) had significantly higher respiratory symptoms score (18.5 vs 6, P<0.01) and Rint expressed as absolute values (kPa l(-1)) and z-scores (1.33 vs 1.16 and 1.42 vs 1.0, P<0.01), respectively. CONCLUSION: Rint measurement is feasible in prematurely born children of preschool age in the ambulatory setup. Preschool children with severe CLD may be identified from preterm children without CLD by increased Rint that may be used as a screening tool and as an outcome measure for interventions.

The secretin G-protein-coupled receptor family: teleost receptors.

Twenty-one members of the secretin family (family 2) of G-protein-coupled receptors (GPCRs) were identified via directed cloning and data-mining of the Fugu Genome Consortium database, representing the most comprehensive description of secretin GPCRs in a teleost fish to date. Duplicated genes were identified for many of the family members, namely the receptors for pituitary adenylate cyclase-activating polypeptide (PACAP)/vasoactive intestinal peptide (VIP), calcitonin, calcitonin gene-related peptide (CGRP), growth hormone releasing hormone (GHRH), glucagon receptor/glucagon-like peptide (GLP) and parathyroid hormone-related peptide (PTHrP)/PTH. Mining of other teleost genomes (zebrafish and Tetraodon) revealed that the duplicated genes identified in the Takifugu genome were also present in these fish. Additional database searching of the Escherichia coli, yeast, Drosophila, Caenorhabditis elegans and Ciona genomes revealed that the family 2 of GPCRs were only present in the multicellular organisms. Orthologues of all the human secretin receptors were identified with the exception of secretin itself. Additional database searches in the Fugu Genome Consortium database also failed to reveal a secretin ligand and so it is hypothesised that both the receptor and the ligand evolved after the divergence of teleost/tetrapod lineages. Phylogenetic analysis at both the protein and the DNA level provided strong support for each of the individual receptor family groupings, but weak support between groups, making evolutionary inferences difficult. A more critical analysis of the PACAP/VIP receptor family confirmed previous hypotheses that the vasoactive intestinal peptide receptor (VPAC(1)R) gene is the ancestral form of the receptor.

Skin-prick testing as a diagnostic aid for childhood asthma.

Diagnosing asthma is problematic when based solely on reported symptoms. The purpose of this study was to evaluate skin-prick testing as a diagnostic aid for asthma in children. Skin-prick testing (SPT) was undertaken in children aged 2-10 years with either no history of wheeze (n = 149) or recent doctor-observed wheeze which responded to treatment with a bronchodilator, the "gold standard" (n = 164). Children with moderate or severe asthma were excluded. SPT positivity increased sharply at age 5 years in wheezers. Data were therefore divided into two age groups: 2- < 5 years (57 controls, 97 wheezers) and 5-10 years (92 controls, 67 wheezers). The sensitivity, specificity, and likelihood ratios of SPT positivity for wheeze were 32%, 89%, and 2.9, respectively, in the younger children, and 82%, 85%, and 5.5, respectively, in the older children. For a prevalence of 30% for asthma, the positive predictive values of a positive SPT were 55% and 70% for the younger and older age groups, respectively. The test characteristics of SPT for helping diagnose asthma in schoolchildren are good. The prevalence of wheeze in preschool children is high, and so SPT should be helpful even in this group. We suggest that clinicians consider skin-prick testing as a diagnostic aid for asthma.

Following basal stem rot in young oil palm plantings.

The PCR primer GanET has previously been shown to be suitable for the specific amplification of DNA from Ganoderma boninense. A DNA extraction and PCR method has been developed that allows for the amplification of the G. boninense DNA from environmental samples of oil palm tissue. The GanET primer reaction was used in conjunction with a palm-sampling programme to investigate the possible infection of young palms through cut frond base surfaces. Ganoderma DNA was detected in frond base material at a greater frequency than would be expected by comparison with current infection levels. Comparisons are made between the height of the frond base infected, the number of frond bases infected, and subsequent development of basal stem rot. The preliminary results suggest that the development of basal stem rot may be more likely to occur when young lower frond bases are infected.

Ganoderma diseases of perennial crops in India--an overview.

The species of Ganoderma recorded from India as causing diseases of perennial crops are listed, and their host range and taxonomy discussed. Four new hosts of G. lucidum are also reported. A decline in productivity and the death of trees are the main economic impacts due to Ganoderma diseases, and the fungus is identified as a serious pathogen of cash crops, forest plantations and trees in natural forests in the country. Ganoderma diseases have been recorded on 144 hosts in India, the major pathogens being G. lucidum and G. applanatum. G. lucidum has been recorded on 91 hosts, and appears to cause the most widespread diseases. Identification has largely been made from morphological and cultural characters, and the names currently in use should therefore be treated with caution. Cultural methods of disease control are largely inefficient in minimising inoculum pressure and in reducing the disease incidence. Chemical methods in combination with soil amendments form short-term solutions for managing the disease and improving productivity. The immediate priorities for developing an efficient management system for Ganoderma diseases in India are: (1) a thorough understanding of the etiology and epidemiology of the diseases on different hosts, (2) clarifying current ambiguity in species names, (3) assessing the inter-relationships between populations of Ganoderma on different hosts and (4) developing tools for early detection of diseases in important crops.

Diagnostic accuracy of bronchodilator responsiveness in wheezy children.

BACKGROUND: The aim of this study was to evaluate the diagnostic accuracy of bronchodilator responsiveness for asthma in 5-10 year old children. METHODS: Spirometric measurements were made in 142 children (58 wheezers) before and after 400 mug inhaled salbutamol. RESULTS: On a receiver operating characteristic curve, a 9% increase in predicted forced expiratory volume in 1 second was the cut off point that provided an acceptable balance of sensitivity and specificity for previous wheeze. This figure was 50% (95% CI 38 to 62) sensitive and 86% (95% CI 78 to 92) specific for detecting previous wheeze and multiplied the initial odds in favour of wheeze by a factor of 3.6 (95% CI 2.0 to 6.3). DISCUSSION: With an estimated pretest probability of wheeze of 10% in the community and 50% in a specialist clinic, the positive predictive values are 29% and 78%, respectively, for a 9% change. The value of bronchodilator responsiveness testing depends on the prevalence of wheeze in the population in which it is to be used.

Molecular relatedness within the "Ascochyta pinodes-complex".

The morphological, molecular and pathological characters of 34 isolates identified as Ascochyta pinodes, A. phaseolorum, and Phoma medicaginis var. pinodella were determined. Initial molecular findings, based on PCR fragment size and RFLPs of sections of the mitochondrial genome, the rRNA gene cluster and the beta-tubulin gene complex, were further investigated by sequencing of the total rRNA ITS regions. Isolates were homogenous for all characteristics determined, with the exception of conidial size and septation, and RFLPs of AT rich (presumptive mitochondrial) DNA. The results suggest that the isolates represent host preferential forms of a single taxon, and this finding is discussed in relation to the known teleomorphs and reference material of the species studied.

Repeatability of airway resistance measurements made using the interrupter technique.

BACKGROUND: To be able to interpret any measurement, its repeatability should be known. This study reports the repeatability of airway resistance measurements using the interrupter technique (Rint) in children with and without respiratory symptoms. METHODS: Children aged 2-10 years who were healthy, had persistent isolated cough, or who had previous wheeze were studied. On the same occasion, three Rint measurements were made 15 minutes apart, before and after placebo and salbutamol given in random order. Results from those given placebo first were analysed for within-occasion repeatability. Between-occasion repeatability measurements were made 2-20 weeks apart (median 3 weeks). RESULTS: For 85 pairs of measurements before and after placebo the limits of agreement were 20% expected resistance and were unaffected by age or health status. The change in resistance following bronchodilator in one of 18 healthy children, 12 of 28 with cough, and 22 of 39 with wheeze exceeded this threshold. For between-occasion measurements the limits of agreement were 32% in 72 healthy subjects, 49% in 57 with cough, and 53% in 95 with previous wheeze. CONCLUSION: The measurement of airways resistance by the interrupter technique is clinically meaningful when change following an intervention such as the administration of bronchodilator is greater than its within-occasion repeatability. Between-occasion repeatability is too poor to judge change confidently.

Airway resistance measured by the interrupter technique: normative data for 2-10 year olds of three ethnicities.

BACKGROUND AND AIMS: The measurement of airway resistance using the interrupter technique (R(int)) is feasible in preschool children and other subjects unable to undertake spirometry. This makes it potentially useful for the measurement of lung function in these groups. Commercial devices use different algorithms to measure pressure and flow from which R(int) is derived. This study provides normative values for British children using devices from a single manufacturer. METHODS: R(int) was measured in 236 healthy children of three ethnic groups (Afro-Caribbean and black African, Bangladeshi, and white British) aged 2-10 years using Micro Medical devices. Software in the devices calculated R(int) from pressure measured by the two point, back extrapolation method from the pressure transient during valve closure, with flow measured just before valve closure. RESULTS: R(int) is related to both age and height, but when age is allowed for there is not a significant relation with height. Neither gender nor any of the ethnicities studied was significantly related to R(int). DISCUSSION: These measurements in healthy children using this technique may be used as reference data for similar populations.

Phenotypic degeneration occurs during sector formation in Metarhizium anisopliae.

AIMS: The formation of sectors was observed during subculturing of an isolate of the entomopathogenic fungus Metarhizium anisopliae, a fungus used for biological control of insect pests. The aim of the investigation was to establish whether sector formation was accompanied by changes in physiological characters. METHODS AND RESULTS: Four degenerative morphological states, with reduced sporulation capacity, were characterized. Subcultures were taken from each sector and four new culture lines established. The new lines were further subcultured every 21 d. A physiological assessment of each line was undertaken after 42 d using TLC of secondary metabolites and fluorogenic enzyme tests. Full sporulation capacity was not regained on subculture, although some cultures recovered partially. Changes in secondary metabolite profiles and the loss in detection of activity of specific enzymes were observed. CONCLUSIONS: Sector formation was frequently accompanied by changes in the ability to produce secondary metabolites and enzymes. SIGNIFICANCE AND IMPACT OF THE STUDY: The results illustrate the importance of maintaining the stability of important cultures during routine subculture. The consequences could have significant implications if degenerate cultures are used as inocula for liquid fermentation cultures or industrial scale production.

Airway resistance measured by the interrupter technique: expiration or inspiration, mean or median?

The measurement of airway resistance by the interrupter technique (Rint) needs standardization. Should measurements be made be during the expiratory or inspiratory phase of tidal breathing? In reported studies, the measurement of Rint has been calculated as the median or mean of a small number of values, is there an important difference? Subjects were 2.5-5.0 yrs (median 4.0 yrs) who had previous respiratory symptoms. The Rint in expiration (RintE) and inspiration (RintI) pre and postsalbutamol, the coefficient of variation (CV) of values contributing to measurements, and bronchodilator responsiveness(BDR) in both phases were compared. Measurements using median and mean were compared. RintE was higher than RintI by 4% (p < 0.01). The CV of values making up RintE and RintI, and BDR measured in expiration and inspiration were similar. The median difference between means and medians of values making up measurements was 0.6% (range -6-11%). RintE has been shown to be consistently greater then RintI but the difference in this study is small. It is suggested that one or the other is chosen as the standard. In the present data the mean of a set of values contributing to a measurement was not significantly different from the median. However, the use of the median has been recommended since it is less affected by possible outlying values such as might be included by fully automated equipment.

Lung function tests for pre-school children.

There are a few devices for measuring lung function in pre-school children. Neither the interrupter technique nor the forced oscillation techniques have been standardised. We highlight some of the issues around the measurement of airway resistance using the interrupter technique and emphasise that, as with other lung function measurements, operators should have a proper understanding of the methods before they can be applied. Both methods for measuring airway resistance have potential for clinical and research application.

Bronchodilator responsiveness and atopy in 5-10-yr-old coughers.

Cough-variant asthma is considered by some to be an asthma phenotype. Bronchodilator responsiveness (BDR) is an undisputed feature of asthma. Of school-aged wheezers, 90% are atopic. Are school-aged coughers who demonstrate BDR also atopic? If so, then it would be reasonable to reserve the diagnosis cough-variant asthma for this particular group. Airway resistance was measured by the interrupter technique (Rint) before and after salbutamol in controls (n=73), coughers (n=63) and previous wheezers (n=63) aged 5-10 yrs. Immunoglobulin (Ig)-E was measured in coughers and wheezers. BDR was expressed as the ratio baseline:post-salbutamol Rint. Groups were of similar age (mean 6.7, range 5-9.9 yrs). Geometric mean baseline Rint was similar in controls and coughers (0.66 and 0.68 kPa x L(-1) x s), but the baseline Rint for wheezers (0.73 kPa x L(-1) s) was greater than that for controls (p=0.05) but not significantly different from coughers (p=0.17). Geometric mean BDR in coughers was 1.22, controls 1.13 and wheezers 1.30 (p=0.01 for coughers and controls; p=0.08 for coughers and wheezers; p<0.001 for controls and wheezers). IgE was lower in coughers than wheezers (geometric means 36 and 364 International Units (IU) x L(-1), p<0.001) and was unrelated to BDR in both groups. In summary, atopy, and not bronchodilator responsiveness, distinguishes groups of coughers from groups of wheezers. A diagnosis of cough-variant asthma cannot be reserved for even those school-aged coughers, who demonstrate bronchodilator responsiveness.

Developing cryopreservation protocols to secure fungal gene function.

Protocols that involve a freezing process are frequently used for the preservation of fungi, but there have been few studies to assess the physiological and genetic stability of isolates after preservation. In this study, the effects of cryopreservation and lyophilisation on the viability, physiology and genetic stability of isolates of Metarhizium anisopliae, Fusarium oxysporum and Serpula lacrymans were examined. It was found that preservation regime influenced the characters of some of the test isolates. Secondary metabolite profiles and extracellular enzyme production can be affected by preservation. Polymorphism's were detected after PCR fingerprinting in replicates of 2 isolates of Metarhizium that had been preserved by cryopreservation and lyophilisation. The results indicated a need to improve existing preservation protocols. Research is being undertaken to develop novel cryopreservation regimes for recalcitrant micro-organisms. Biochemical and molecular techniques will be used to assess the effectiveness of preservation.