Pseudomonas syringae pv. coryli, the Causal Agent of Bacterial Twig Dieback of Corylus avellana

ABSTRACT Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by a polyphasic approach. All strains were assessed by fatty acids analysis and repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX and ERIC primer sets. Representative strains also were assessed by sequencing the 16S rDNA and hrpL genes, determining the presence of the syrB gene, testing their biochemical and nutritional characteristics, and determining their pathogenicity to hazelnut and other plants species or plant organs. Moreover, they were compared with reference strains of other phytopathogenic pseudomonads. The strains from hazelnut belong to Pseudomonas syringae (sensu latu), LOPAT group Ia. Both fatty acids and repetitive-sequence-based PCR clearly discriminate such strains from other Pseudomonas spp., including P. avellanae and other P. syringae pathovars as well as P. syringae pv. syringae strains from hazelnut. Also, the sequencing of 16S rDNA and hrpL genes differentiated them from P. avellanae and from P. syringae pv. syringae. They did not possess the syrB gene. Some nutritional tests also differentiated them from related P. syringae pathovars. Upon artificial inoculation, these strains incited severe twig diebacks only on hazelnut. Our results justify the creation of a new pathovar because the strains from hazelnut constitute a homogeneous group and a discrete phenon. The name of P. syringae pv. coryli is proposed and criteria for routine identification are presented.     

Epitope identification and <Emphasis Type="Italic">in silico</Emphasis> prediction of the specificity of antibodies binding to the coat proteins of <Emphasis Type="Italic">Potato Virus Y</Emphasis> strains

A phage library containing 2.7 × 10⁹ randomly expressed peptides was used to determine the epitopes of three monoclonal antibodies that bind to the coat protein of potato virus Y. Construction of the consensus sequences for the peptides obtained after three selection rounds indicated that each antibody recognized a different epitope located within the first 50 N-terminal amino acids of the coat protein. The location of the epitopes was confirmed by heterologous expression of the N-terminal part of the coat protein in Escherichia coli, and, subsequently, by performing an immunological test with the three antibodies. The accuracy of the phage library was demonstrated by predicting in silico the cross-reactivity of the three antibodies with other potyvirus family members. ELISA and in silico predictions revealed the same results in almost every case. The potential of peptide phage libraries to optimize the use of antibodies in plant virology is discussed.     

Genotyping of Mycoplasma gallisepticum and M. synoviae by Amplified Fragment Length Polymorphism (AFLP) analysis and digitalized Random Amplified Polymorphic DNA (RAPD) analysis

Mycoplasma gallisepticum (MG) and M. synoviae (MS) are the cause of considerable economic losses in the poultry industry. Molecular differentiation of avian Mycoplasma strains may be helpful in tracing infections and in the evaluation of implemented intervention strategies. Amplified Fragment Length Polymorphism (AFLP) has shown to be a powerful typing technique but the application for poultry Mycoplasma strains is very limited. The aim of this study was to evaluate the reproducibility and discriminatory power of AFLP HindIII/HhaI and AFLP BglII/Mfel for the inter- and intraspecies differentiation of avian mycoplasmas and to compare these test characteristics with digitalized Random Amplified Polymorphic DNA (RAPD) analysis. The reproducibility of RAPD, AFLP HindIII/HhaI and AFLP BglII/Mfel was 50-100, 97-98 and 86-99%, respectively. RAPD and both AFLP enzyme combinations were able to differentiate between five avian Mycoplasma species. For AFLP, five MG and four MS clusters could be identified. The phylogenetic tree for both enzyme combinations was comparable. For RAPD, four MG clusters could be identified. For MS, however, due to the poor reproducibility of the RAPD technique, no clear genogroups could be identified. On basis of the results of this study it can be concluded that AFLP is a powerful technique for the genotyping of avian mycoplasmas and that, although AFLP HindIII/HhaI generated patterns with less fragments, the final results showed homologous results.     

The impact of daily Duddingtonia flagrans application to lactating ewes on gastrointestinal nematodes infections in their lambs in the Netherlands

Two experiments were performed in 2002 and 2003 to evaluate the effect of biological control of gastrointestinal nematodes in sheep through the daily feeding of 500,000 chlamydospores of Duddingtonia flagrans/kg bodyweight to lactating ewes during the first 9 weeks with their young lambs on pasture. In both experiments four groups of eight ewes and their April-borne lambs were used. They were turned out on four separate plots (plots A) at the beginning of May, moved to similar separate plots after 3 (plots B) and 6 weeks (plots C), respectively, and weaning occurred after 9 weeks. In both experiments, two groups were fed spores daily while the two other groups served as controls. The effect of D. flagrans application was evaluated through faecal egg counts of ewes and lambs, the yield of faecal cultures in ewes, pasture larval counts and worm counts of lambs and tracer lambs. The results demonstrated no effect of D. flagrans application during the first 5 (2002) or 4 (2003) weeks. Subsequently, fungus application strongly reduced the yield in faecal cultures of the ewes. This was, however, not reflected in the pasture larval counts, but lower worm burdens were observed in tracer lambs of 'treated' plots C in 2002 than on those of 'control' plots. In 2003 worm burdens in 'treated' lambs returned to plots B were lower than those of 'control' lambs and a tendency for the same was observed for plots C. However, in all groups, lambs and tracer lambs developed severe haemonchosis.     

Early reactivation of European rivers during the last deglaciation

During the Last Glacial Maximum, the sea-level lowstand combined with the large extent of the Fennoscandian and British ice sheets led to the funneling of European continental runoff, resulting in the largest river system that ever drained the European continent. Here, we show an abrupt and early reactivation of the European hydrological cycle at the onset of the last deglaciation, leading to intense discharge of the Channel River into the Bay of Biscay. This freshwater influx, probably combined with inputs from proglacial or ice-dammed lakes, dramatically affected the hydrology of the region, both on land and in the ocean.     

Highly conserved protective epitopes on influenza B viruses

Identification of broadly neutralizing antibodies against influenza A viruses has raised hopes for the development of monoclonal antibody-based immunotherapy and "universal" vaccines for influenza. However, a substantial part of the annual flu burden is caused by two cocirculating, antigenically distinct lineages of influenza B viruses. Here, we report human monoclonal antibodies, CR8033, CR8071, and CR9114, that protect mice against lethal challenge from both lineages. Antibodies CR8033 and CR8071 recognize distinct conserved epitopes in the head region of the influenza B hemagglutinin (HA), whereas CR9114 binds a conserved epitope in the HA stem and protects against lethal challenge with influenza A and B viruses. These antibodies may inform on development of monoclonal antibody-based treatments and a universal flu vaccine for all influenza A and B viruses.     

Livestock-associated MRSA prevalence in veal calf production is associated with farm hygiene, use of antimicrobials, and age of the calves

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is highly prevalent in pork and veal production chains. In this study, we used data from a cross-sectional survey on 2151 calves from 102 veal calf farms to identify potential risk factors, with the goal of reducing MRSA prevalence by developing intervention strategies. Overall, calves from rose veal farms had a lower risk of LA-MRSA carriage than calves from white veal farms. Data were analysed separately for white and rose veal calves, because management systems of the two production chains were largely different. Group treatment with antimicrobials appeared to be a risk factor for MRSA carriage in white veal calves in univariate analyses, but was not included in the final multiple regression model that included age of the calves and rodent control. Number of start treatment days was positively associated with LA-MRSA carriage in rose veal calves, and was the only risk factor selected for the final multiple regression model for this group. Interpretation of the results from this cross-sectional study is complicated by the strong correlation between antimicrobial use, LA-MRSA carriage and age of the calves. Other age-related factors may be more influential. However, taken together these findings emphasize the need for prudent use of antimicrobials, and point to improvement of farm hygiene as a control measure.     

Radiographic findings of the medial humeral epicondyle in 200 canine elbow joints

Objectives: To determine the frequency and radiographic aspect of medial humeral epicondylar lesions as a primary or concomitant finding and to evaluate the association with osteoarthritis. Methods: Medical records of dogs diagnosed with elbow lameness were reviewed. Inclusion criteria for this study were a complete clinical examination, a complete set of digital radiographs and a final diagnosis made by computed tomography or magnetic resonance imaging and arthroscopy. Changes of the medial humeral epicondyle were recorded and correlated with the radiographic osteoarthritis and final diagnosis. Results: Eighty of the 200 elbows showed changes of the medial humeral epicondyle. In 12 of these 80 elbows, changes of the medial epicondyle were the only findings within the joint, and these elbows were diagnosed with primary flexor enthesopathy. In the remaining 68 elbows, other concomitant elbow pathologies were found. In those cases of concomitant epicondylar changes, high grades of osteoarthritis were recorded, while most elbows with primary flexor enthesopathy showed a low grade of osteoarthritis. Clinical significance: Changes of the medial humeral epicondyle are often considered clinically unimportant and are regarded as an expression of osteoarthritis. This study showed the relatively frequent presence of epicondylar changes of which the majority were considered concomitant to a primary elbow problem. If changes of the medial humeral epicondyle are the only pathologic finding (primary flexor enthesopathy) they should be considered as the cause of lameness and not as a sign of osteoarthritis.