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    ABSTRACT: The full-length cDNA of four Ofapn aminopeptidases were cloned and sequenced from susceptible and resistant Ostrinia furnacalis strains. Four sequences were identified as APN because they shared the common structural features with APN from Lepidoptera, including the signal peptide, GPI anchor signal, the zinc binding/gluzincin motif HEX2HX18E and the gluzincin aminopeptidase motif GAMEN. Compared with APN sequences from the susceptible strain, there were 9, 5, 10 and 12 amino acid variations in the deduced protein sequences from the resistant strain. There were also differences in mRNA expression of the four Ofapn genes between resistant and susceptible O. furnacalis strains.
    Journal of Invertebrate Pathology 11/2013; 115. DOI:10.1016/j.jip.2013.11.001
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    ABSTRACT: American Foulbrood (AFB) caused by Paenibacillus larvae is the most damaging bacterial brood disease of the honey bee (Apis mellifera), causing colony deaths on all continents where honey bees are managed. AFB has been a persistent problem in the UK for over 70 years, with a fluctuating number of cases discovered annually. Once diseased colonies are identified, they are destroyed to reduce pathogen spread. We investigated the pattern of AFB cases recorded over the period 1994 to 2012 using spatial statistical approaches with a view to identifying the nature of spread across England and Wales. Our results indicated that AFB exhibits significant spatial aggregation at distances from 10-30 km with aggregations lasting between 1 and 5 year periods. Kernel smoothing indicated areas of elevated relative risk in different years and these were further detailed by spatial scan statistics. We identified disease clusters and successfully estimated their size, location and duration. The majority of clusters did not persist in all years indicating that management measures may lead to localised extinction of the disease. Whilst less common, persistent clusters likely indicate potential endemic or exotic risk points. The application of robust epidemiological approaches to improve the control of AFB are discussed.
    Environmental Microbiology 10/2013; 16(12). DOI:10.1111/1462-2920.12292
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    ABSTRACT: Three acidophilic actinobacteria, isolates LSCA2, FGG8 and HSCA14T, recovered from spruce litter were examined using a polyphasic approach. Chemotaxonomic and morphological properties of the isolates were found to be consistent with their classification in the genus Streptacidiphilus. The isolates were shown to have identical 16S rRNA gene sequences and were most closely related to Streptacidiphilus neutrinimicus DSM 41755T (99.9 % similarity). However, DNA:DNA relatedness between isolate HSCA14T and the type strain of S. neutrinimicus was found to be low at 44.0 (±14.1) %. A combination of phenotypic features, including degradative and nutritional characteristics were shown to distinguish the isolates from their nearest phylogenetic neighbours. Data from this study show that the isolates form a novel species in the genus for which the name S. hamsterleyensis sp. nov. is proposed. The type strain is HSCA 14T (=DSM 45900T = KACC 17456T = NCIMB 14865T). Electronic supplementary material The online version of this article (doi:10.1007/s10482-013-0015-1) contains supplementary material, which is available to authorized users.
    Antonie van Leeuwenhoek 08/2013; 104(6). DOI:10.1007/s10482-013-0015-1
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    ABSTRACT: Low cost, simple approaches leading to enhanced numbers of viable, mature corals on reefs are prerequisite to active reef rehabilitation at even modest spatial scales. Mass culture of coral larvae to settlement, utilising improved knowledge of major coral spawning events, promises to be relatively straightforward, but very significant mortality in the early post-settlement period remains a major hurdle. This study was conducted to examine the effect of herbivore exclusion on the survival of 6 week old coral spat of Acropora tenuis (Dana, 1846) reared ex situ at a site in north-western Philippines. Coral spat were placed on the reef approximately 6 weeks after settlement in three treatments, caged, open-sided cage and no cage at two depths (4 m and 9 m). Mean survival of coral spat was significantly higher at the deep sites compared to the shallow sites. Among treatments, survival was significantly lower in the uncaged treatment (4.7% ± 2.6% and 10.5% ± 4.5%, mean ± SE in shallow and deep respectively) compared to the open-sided cage (18.6% ± 5.0% and 22.5% ± 7.1%) and fully caged treatment (17.0% ± 4.5% and 33.0% ± 6.0%) after 3 months. The results indicate that removal of coral spat by grazers may have reduced survival in the uncaged treatment, although the fact that survivorship was not significantly reduced in the open-sided cage treatments suggests that the presence of the cage also had some effect on survival. It is possible that the open-sided cage prevented access by larger fish that may have actively removed coral spat or that shading provided by the cage enhanced spat survival. Further research is needed to see if survivorship in cages decreases at a later stage due to overgrowth by other biota and whether survivorship is enhanced if spat are settled on more complex surfaces that provide refuge from grazers. This study demonstrates that using cages to exclude herbivores and corallivores and/or to provide shading may be beneficial to survival during the early stages when rearing corals in situ for reef rehabilitation.
    Journal of Experimental Marine Biology and Ecology 08/2013; DOI:10.1016/j.jembe.2010.08.003
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    ABSTRACT: Benthic algae are associated with coral death in the form of stress and disease. It's been proposed that they release exudates, which facilitate invasion of potentially pathogenic microbes at the coral-algal interface, resulting in coral disease. However, the original source of these pathogens remains unknown. This study examined the ability of benthic algae to act as reservoirs of coral pathogens by characterizing surface associated microbes associated with major Caribbean and Indo-Pacific algal species/types and by comparing them to potential pathogens of two dominant coral diseases: White Syndrome (WS) in the Indo-Pacific and Yellow Band Disease (YBD) in the Caribbean. Coral and algal sampling was conducted simultaneously at the same sites to avoid spatial effects. Potential pathogens were defined as those absent or rare in healthy corals, increasing in abundance in healthy tissues adjacent to a disease lesion, and dominant in disease lesions. Potentially pathogenic bacteria were detected in both WS and YBD and were also present within the majority of algal species/types (54 and 100% for WS and YBD respectively). Pathogenic ciliates were associated only with WS and not YBD lesions and these were also present in 36% of the Indo-Pacific algal species. Although potential pathogens were associated with many algal species, their presence was inconsistent among replicate algal samples and detection rates were relatively low, suggestive of low density and occurrence. At the community level, coral-associated microbes irrespective of the health of their host differed from algal-associated microbes, supporting that algae and corals have distinctive microbial communities associated with their tissue. We conclude that benthic algae are common reservoirs for a variety of different potential coral pathogens. However, algal-associated microbes alone are unlikely to cause coral death. Initial damage or stress to the coral via other competitive mechanisms is most likely a prerequisite to potential transmission of these pathogens.
    PLoS ONE 07/2013; 8(7):e69717. DOI:10.1371/journal.pone.0069717
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    ABSTRACT: We demonstrate the first application of synthetic RNA gene silencers in Streptomyces coelicolor A3(2). Peptide nucleic acid and expressed antisense RNA silencers successfully inhibited actinorhodin production. Synthetic RNA silencing was target-specific and is a new tool for gene regulation and metabolic engineering studies in Streptomyces.
    PLoS ONE 06/2013; 8(6):e67509. DOI:10.1371/journal.pone.0067509
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    ABSTRACT: The taxonomic position of three acidophilic actinobacteria, strains FGG38, FGG39 and FSCA67(T), isolated from the fermentation litter layer of a spruce forest soil was established using a polyphasic approach. The strains were shown to have chemotaxonomic and morphological properties consistent with their classification in the genus Streptacidiphilus and formed a distinct phyletic line in the Streptacidiphilus 16S rRNA gene tree being most closely related to Streptacidiphilus albus DSM 41753(T) (99.4 % similarity). DNA:DNA relatedness data showed that isolate FSCA67(T) and the type strain of S. albus belonged to markedly distinct genomic species. The isolates had many phenotypic properties in common and were distinguished readily from their closest phylogenetic neighbours in the Streptacidiphilus gene tree using a broad range of these features. Based on the combined genotypic and phenotypic data the three isolates are considered to represent a new Streptacidiphilus species. The name Streptacidiphilus durhamensis sp. nov. is proposed for this taxon with isolate FSCA67(T) (=DSM 45796(T) = KACC 17154(T) = NCIMB 14829(T)) as the type strain.
    Antonie van Leeuwenhoek 05/2013; 104(2). DOI:10.1007/s10482-013-9938-9
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    ABSTRACT: SUMMARY The aim of this study was to investigate spatial variation in risk of hospitalization in childhood pneumonia and empyema in the North of England and associated risk factors. Data on childhood (0-14 years) hospital admissions with a diagnosis pneumonia or empyema were linked to postcode districts. Bayesian conditional autoregressive models were used to evaluate spatial variation and the relevance of specific spatial covariates in an area-based study using postcode as the areal unit. There was a sixfold variation in the risk of hospitalization due to pneumonia across the study region. Variation in risk was associated with material deprivation, Child Well-being Index (CWI) health domain score, number of children requiring local authority support, and distance to hospital. No significant spatial variation in risk for empyema was found.
    Epidemiology and Infection 05/2013; 142(02):1-11. DOI:10.1017/S0950268813001015
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    ABSTRACT: Background and AimsDespite the selective pressure slugs may exert on seedling recruitment there is a lack of information in this context within grassland restoration studies. Selective grazing is influenced by interspecific differences in acceptability. As part of a larger study of how slug-seedling interactions may influence upland hay meadow restoration, an assessment of relative acceptability is made for seedlings of meadow plants to the slug, Deroceras reticulatum.Methods Slug feeding damage to seedling monocultures of 23 meadow species and Brassica napus was assessed in microcosms over 14 d. The severity and rate of damage incurred by each plant species was analysed with a generalized additive mixed model. Plant species were then ranked for their relative acceptability.Key ResultsInterspecific variation in relative acceptability suggested seedlings of meadow species form a hierarchy of acceptability to D. reticulatum. The four most acceptable species were Achillea millefolium and the grasses Holcus lanatus, Poa trivialis and Festuca rubra. Trifolium pratense was acceptable to D. reticulatum and was the second highest ranking forb species. The most unacceptable species were mainly forbs associated with the target grassland, and included Geranium sylvaticum, Rumex acetosa, Leontodon hispidus and the grass Anthoxanthum odoratum. A strong positive correlation was found for mean cumulative feeding damage and cumulative seedling mortality at day 14.Conclusions Highly unacceptable species to D. reticulatum are unlikely to be selectively grazed by slugs during the seedling recruitment phase, and were predominantly target restoration species. Seedlings of highly acceptable species may be less likely to survive slug herbivory and contribute to seedling recruitment at restoration sites. Selective slug herbivory, influenced by acceptability, may influence community-level processes if seedling recruitment and establishment of key functional species, such as T. pratense is reduced.
    Annals of Botany 04/2013; DOI:10.1093/aob/mct086
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    ABSTRACT: The number and prevalence of coral diseases/syndromes are increasing worldwide. Dark Spot Syndrome (DSS) afflicts numerous coral species and is widespread throughout the Caribbean, yet there are no known causal agents. In this study we aimed to characterise the microbial communities (bacteria and fungi) associated with DSS lesions affecting the coral Stephanocoenia intersepta using nonculture molecular techniques. Bacterial diversity of healthy tissues (H), those in advance of the lesion interface (apparently healthy AH), and three sizes of disease lesions (small, medium, and large) varied significantly (ANOSIM R = 0.052 p<0.001), apart from the medium and large lesions, which were similar in their community profile. Four bacteria fitted into the pattern expected from potential pathogens; namely absent from H, increasing in abundance within AH, and dominant in the lesions themselves. These included ribotypes related to Corynebacterium (KC190237), Acinetobacter (KC190251), Parvularculaceae (KC19027), and Oscillatoria (KC190271). Furthermore, two Vibrio species, a genus including many proposed coral pathogens, dominated the disease lesion and were absent from H and AH tissues, making them candidates as potential pathogens for DSS. In contrast, other members of bacteria from the same genus, such as V. harveyii were present throughout all sample types, supporting previous studies where potential coral pathogens exist in healthy tissues. Fungal diversity varied significantly as well, however the main difference between diseased and healthy tissues was the dominance of one ribotype, closely related to the plant pathogen, Rhytisma acerinum, a known causal agent of tar spot on tree leaves. As the corals' symbiotic algae have been shown to turn to a darker pigmented state in DSS (giving rise to the syndromes name), the two most likely pathogens are R. acerinum and the bacterium Oscillatoria, which has been identified as the causal agent of the colouration in Black Band Disease, another widespread coral disease.
    PLoS ONE 04/2013; 8(4):e62580. DOI:10.1371/journal.pone.0062580
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