ISApl1, a novel insertion element of Actinobacillus pleuropneumoniae, prevents ApxIV-based serological detection of serotype 7 strain AP76

Actinobacillus pleuropneumoniae, a gram-negative rod of the Pasteurellaceae family, causes pleuropneumonia in pigs. Establishing A. pleuropneumoniae free herds is difficult due to the occurrence of persistently infected animals. The ApxIV toxin is expressed by A. pleuropneumoniae in vivo and an ELISA based on the toxin is used to detect infection and to differentiate between infected and vaccinated animals. In this study, we have identified a 1070bp insertion element of the IS30 family, designated ISApl1, in the A. pleuropneumoniae serotype 7 strain AP76. ISApl1 contains a 924bp ORF encoding a transposase, which is flanked by 27bp inverted repeats showing six mismatches. We investigated the occurrence of ISApl1 in other A. pleuropneumoniae strains, and its possible interference with virulence associated factors. Four insertion sites were identified in AP76: within the apxIVA toxin ORF, within a putative autotransporter adhesin ORF, upstream of a capsular polysaccharide biosynthesis gene cluster, and downstream of a beta-lactamase gene. ISApl1 is also present in some serotype 7 field isolates, but not in reference or field strains of other serotypes. In A. pleuropneumoniae AP76, the transposase gene is transcribed in vitro. The insertion in the apxIVA toxin gene remains stable after animal passage. Since this insertion should disrupt toxin expression, we tested 7 pigs infected with AP76 at day 21 post-infection. All were negative in the ApxIV ELISA but four out of seven were positive in an ApxII toxin ELISA. These results show that insertion elements can affect the detection of A. pleuropneumoniae infected animals.     

Understanding and valuing conflicts in marine protected areas: The best way to develop innovations?

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Simplified performance assessment methodology for addressing soil quality of nature-based solutions

Journal of Soils and Sediments - In urban areas, soil functions are deeply impacted by all human activities, e.g., water infiltration, carbon storage, and chemical substances degradation potential....     

The impact of body site, topical melatonin and brushing on hair regrowth after clipping normal Siberian Husky dogs

The aims of this study were to determine the impact of body site, vigorous brushing and topical melatonin treatment on hair regrowth after clipping normal dogs. Siberian Husky dogs were randomly assigned to three groups of eight dogs each. All dogs had the lumbosacral region and both lateral thighs clipped. The left thigh and lumbosacral area received no treatment and were compared in all 24 dogs. Eight dogs had the right thigh treated with 0.1% melatonin twice daily for 2 months, and hair regrowth was compared with the left thigh. Eight dogs had the right thigh brushed twice daily for 2 months, and hair regrowth was compared with the left thigh. Eight dogs had neither thigh treated. Hairs were plucked before and 2 months postclipping, and the proportion of hair growth from the original length was calculated and compared as described above. Biopsy samples were collected before and after treatment to determine if brushing induced dermal inflammation and melatonin increased the proportion of anagen follicles. Proportionally, left thigh hairs were significantly longer compared to lumbosacral hairs 2 months postclipping. No significant differences in hair regrowth were noted between the nontreated thigh and the thigh treated with melatonin or brushed. No significant difference in dermal inflammation was noted before and after brushing. No significant differences were observed in the proportion of anagen follicles before and after topical melatonin treatment. Our results showed that the hairs in the lumbosacral region were proportionally shorter than lateral thigh hairs 2 months postclipping. Moreover, topical melatonin and brushing had no impact on hair regrowth after clipping normal dogs.     

Acibenzolar-S-methyl Induces the Accumulation of Defense-related Enzymes in Apple and Protects from Fire Blight

Acibenzolar-S-methyl (Novartis) is a chemical inducer of systemic acquired resistance in several annual plants. The ability of this novel chemical to induce resistance was studied in a perennial plant (apple) affected by fire blight caused by Erwinia amylovora. Acibenzolar-S-methyl (100 and 200mg/l active ingredient) protected Golden Delicious seedlings, scions and trees from artificial infection when applied before inoculation. The protection of apple seedlings was similar to the protection obtained with the standard for fire blight control, plantomycin (100mg/l streptomycin sulfate), applied immediately before inoculation. The mean levels of control in scions in the greenhouse and in trees in orchards were approximately 69% and 50%, respectively. The protection of apple seedlings was constantly associated with the activation of two families of defense-related enzymes, peroxidases and -1,3-glucanases. Accumulation of both enzymes was induced locally in treated leaves and systemically, especially for -1,3-glucanases, in upper untreated leaves, and was sustained for at least 17 days. These results suggest that acibenzolar-S-methyl promotes induced systemic resistance in apple by increasing defense-related compounds. This chemical could provide a new approach of control of fire blight but its practical use needs further investigation.     

Metabolism of [14C]-2,4-dichlorophenol in edible plants

Several 2,4-dichlorophenoxyacetic acid (2,4-D)-sensitive plants have been modified by genetic engineering with tfdA gene to acquire 2,4-D tolerance. The expression product of this gene degrades 2,4-D to 2,4-dichlorophenol (DCP), which is less phytotoxic but could cause a problem of food safety. After a comparison of 2,4-D and DCP metabolism in transgenic 2,4-D-tolerant and wild cotton (Gossypium hirsutum L.), a direct study of DCP metabolism in edible plants was performed. After petiolar uptake of a [U-phenyl-(14)C]-DCP solution followed by a 48 h water chase, aqueous extracts were analysed by high-performance liquid chromatography. Metabolites were thereafter isolated and their structural identities were determined by enzymatic and chemical hydrolyses and mass spectrometry analyses. The metabolic fate of DCP was equivalent to 2,4-D metabolism in transgenic 2,4-D-tolerant cotton. In addition, DCP metabolism was similar in transgenic and wild cotton. The major terminal metabolites were DCP-saccharide conjugates in all species, essentially DCP-(6-O-malonyl)-glucoside or its precursor DCP-glucose. The significance of this metabolic pathway with regard to food safety is discussed.     

Population structure and virulence content of avian pathogenic Escherichia coli isolated from outbreaks in Sri Lanka

Avian pathogenic Escherichia coli (APEC) causes economically significant infections in poultry. The genetic diversity of APEC and phylogenetic relationships within and between APEC and other pathogenic E. coli are not yet well understood. We used multilocus sequence typing (MLST), PCR-based phylogrouping and virulence genotyping to analyse 75 avian E. coli strains, including 55 isolated from outbreaks of colisepticaemia and 20 from healthy chickens. Isolates were collected from 42 commercial layer and broiler chicken farms in Sri Lanka. MLST identified 61 sequence types (ST) with 44 being novel. The most frequent ST, ST48, was represented by only six isolates followed by ST117 with four isolates. Phylogenetic clusters based on MLST sequences were mostly comparable to phylogrouping by PCR and MLST further differentiated phylogroups B1 and D into two subgroups. Genotyping of 16 APEC associated virulence genes found that 27 of the clinical isolates and one isolate from a healthy chicken belonged to highly virulent genotype according to previously established classification schemes. We found that a combination of four genes, ompT, hlyF, iroN and papC, gave a comparable prediction to that of using five and nine genes by other studies. Four STs (ST10, ST48, ST117 and ST2016) contained APEC isolates from this study and human UPEC isolates reported by others, suggesting that these STs are potentially zoonotic. Our results enhanced the understanding of APEC population structure and virulence association.     

C-cadherin ectodomain structure and implications for cell adhesion mechanisms

Cadherins are transmembrane proteins that mediate adhesion between cells in the solid tissues of animals. Here we present the 3.1 angstrom resolution crystal structure of the whole, functional extracellular domain from C-cadherin, a representative "classical" cadherin. The structure suggests a molecular mechanism for adhesion between cells by classical cadherins, and it provides a new framework for understanding both cis (same cell) and trans (juxtaposed cell) cadherin interactions. The trans adhesive interface is a twofold symmetric interaction defined by a conserved tryptophan side chain at the membrane-distal end of a cadherin molecule from one cell, which inserts into a hydrophobic pocket at the membrane-distal end of a cadherin molecule from the opposing cell.     

Identifying kinetic barriers to mechanical unfolding of the T. thermophila ribozyme

Mechanical unfolding trajectories for single molecules of the Tetrahymena thermophila ribozyme display eight intermediates corresponding to discrete kinetic barriers that oppose mechanical unfolding with lifetimes of seconds and rupture forces between 10 and 30 piconewtons. Barriers are magnesium dependent and correspond to known intra- and interdomain interactions. Several barrier structures are "brittle," breakage requiring high forces but small (1 to 3 nanometers) deformations. Barrier crossing is stochastic, leading to variable unfolding paths. The response of complex RNA structures to locally applied mechanical forces may be analogous to the responses of RNA during translation, messenger RNA export from the nucleus, and viral replication.