Temperature, the GacS/GacA system, and host metabolites regulate the type VI secretion system of Pseudomonas syringae pv. phaseolicola

The recently discovered type VI secretion system (T6SS) is implicated in the pathogenic and/or virulence processes of diverse bacteria. The expression pattern of the T6SS differs among different organisms and also depends on several environmental factors. We initiated a study of the conditions that influence T6SS gene expression in Pseudomonas syringae pv. phaseolicola NPS3121. Our results indicate that low temperatures and plant extracts impact the expression of T6SS genes and that the process is subject to regulation by the GacS/GacA two-component system.     

Isolation and genetic characterization of Acidovorax avenae from red stripe infected sugarcane in Northwestern Argentina

Red stripe is a bacterial disease of sugarcane causing important economic losses in Argentina that affects 30 % of the milling stems and consequently the juice quality. In this study, sugarcane leaves exhibiting red stripe symptoms were sampled in the 2008–09 growing season from 13 different sugarcane producing areas of Tucumán and Salta (northwest of Argentina). To achieve the identification and characterization of the causal agent of red stripe, bacterial isolation was performed. Species-specific PCR using Oaf1/Oar1 primers allowed the amplification of a fragment of 550 bp from approximately 50 % of the isolates; 16S rDNA sequences analysis displayed a similarity greater than 99 % with Acidovorax avenae subsp. avenae. By means of RAPD-PCR the presence of at least four different biotypes among the analyzed isolates was detected. Results of pathogenicity test allowed us to confirm A. avenae subsp. avenae as the pathogenic agent for red stripe. This study constitutes the first report on the identification and molecular characterization of this plant pathogen from the Argentina sugarcane production areas. The genetic diversity observed among A. avenae is an important factor to be considered to improve an accurate diagnosis and/or the selection of sugarcane tolerant clones.     

New hosts for Lecanosticta acicola and Dothistroma septosporum in newly established arboreta in Spain

A historical outbreak of needle blight disease was recorded during 2018 to 2019 in plantations of Pinus radiata and Pinus nigra in the North of Spain. The main pathogens involved in this historical outbreak were identified as Lecanosticta acicola and Dothistroma septosporum. Recently, a variety of tree species in three arboreta planted between 2011 and 2013 in the Basque Country as part of the European project REINFFORCE were showing symptoms of needle blight and defoliation. The aim of this study was to determine which pine species were affected with these pathogens. Tree species sampled included several provenances of P. brutia, P. elliottii, P. nigra, P. pinaster, P. pinea, P. ponderosa, P. sylvestris and P. taeda. Using molecular identification methods, Lecanosticta acicola was confirmed infecting Pinus brutia (Provenance: Alexandropolis, Greece and var. eldarica, Crimea) and represents a new host species for this pathogen. Pinus elliottii (Provenance: Georgia, USA) and P. ponderosa (Provenance: Central California, USA) are new host reports of L. acicola for Spain. Dothistroma septosporum was found for the first time on P. brutia (Provenance: Marmaris, Turkey) and P. ponderosa (Provenance: Oregon, USA) in Spain and was also detected infecting P. nigra (Provenance: Sologne Vayrières, France).     

Suppression of avian influenza transmission in genetically modified chickens

Infection of chickens with avian influenza virus poses a global threat to both poultry production and human health that is not adequately controlled by vaccination or by biosecurity measures. A novel alternative strategy is to develop chickens that are genetically resistant to infection. We generated transgenic chickens expressing a short-hairpin RNA designed to function as a decoy that inhibits and blocks influenza virus polymerase and hence interferes with virus propagation. Susceptibility to primary challenge with highly pathogenic avian influenza virus and onward transmission dynamics were determined. Although the transgenic birds succumbed to the initial experimental challenge, onward transmission to both transgenic and nontransgenic birds was prevented.     

Contingent Valuation of Woodland-Owner Private Amenities in Spain,Portugal, and California

Most of the Mediterranean woodlands in Spain, Portugal, and California are managed as agrosilvopastoral enterprises, producing some combination of livestock, wood, cork products, and crops, as well as wildlife habitat and diverse environmental services. Private amenity benefits to landowners have been suggested as an explanation for high land prices and the persistence of such rangeland enterprises despite apparently marginal cash returns. In this study, private amenity values are estimated using a contingent valuation technique in surveys of private woodland owners as part of five case studies, using a design developed to separate landowner amenity income and capital values. Nonindustrial private landowners were asked about the maximum amount of money that they were willing to give up (to pay) before selling their property to invest in more commercially profitable assets, and the proportion of the market price of their woodland that they think is explained by privately consumed amenities. Amenity values were found to be relevant because, in all cases, landowners were willing to pay &spigt; €120 · ha^-1 · yr^-1, at 2002 prices, and attributed &spigt; 30% of land market price to amenities. These values represent an amenity profitability rate &spigt; 2% in all case studies. The data analysis shows some similarities, but mostly divergences, in the different land-simulated and amenity-simulated markets.     

Efficiency of DNA typing methods for detection of smoked paprika "pimenton de la vera" adulteration used in the elaboration of dry-cured Iberian pork sausages

The purpose of this work was to develop a PCR method for the identification of smoked paprika "Pimento?n de la Vera" adulteration with paprika elaborated from varieties of pepper foreign to the la Vera region, in central western Spain. Three autochthonous varieties of pepper, Jaranda, Jariza, and Bola, and the varieties Papri Queen, Papri King, Sonora, PS9794, and Papri Ace, foreign to the La Vera region, were used in the study. Analyses of the ITS and 5.8S rDNA, RAPD-PCR, SSR, and ISSR were tested. RAPD-PCR, SSR, and ISSR analyses allowed differentiation among the varieties of paprika analyzed. There was no difference in the sequence of ITS1-5.8S rDNA-ITS2. In addition, with the RAPD-PCR primers S13 and S22, two molecular markers were obtained of 641 and 704 bp, respectively, which allowed all of the smoked paprika varieties to be differentiated from paprikas elaborated with the five foreign varieties. These two molecular markers were investigated as a basis for detecting the adulteration of smoked paprika with paprika elaborated from foreign varieties of pepper.     

Renibacterium salmoninarum iron‐acquisition mechanisms and ASK cell line infection: Virulence and immune response

Renibacterium salmoninarum is the aetiological agent of bacterial kidney disease (BKD) in salmonid farms. This pathogen possesses at least three iron‐acquisition mechanisms, but the link between these mechanisms and virulence is unclear. Therefore, this study used RT‐qPCR to assess the effects of normal and iron‐limited conditions on iron‐uptake genes controlled by IdeR and related to iron acquisition in Chilean R. salmoninarum strain H‐2 and the type strain DSM20767^T. Further evaluated was the in vitro immune‐related response of the Atlantic Salmon Kidney (ASK) cell line, derived from the primary organ affected by BKD. R. salmoninarum grown under iron‐limited conditions overexpressed genes involved in haemin uptake and siderophore transport, with overexpression significantly higher in H‐2 than DSM20767^T. These overexpressed genes resulted in higher cytotoxicity and an increased immune response (i.e., TNF‐α, IL‐1β, TLR1 and INF‐γ) in the ASK cell line. This response was significantly higher against bacteria grown under iron‐limited conditions, especially H‐2. These observations indicate that iron‐acquisition mechanisms are possibly highly related to the virulence and pathogenic capacity of R. salmoninarum. In conclusion, treatments that block iron‐uptake mechanisms or siderophore synthesis are attractive therapeutic approaches for treating R. salmoninarum, which causes significant aquaculture losses.     

Culture possibilities of the razor clam Ensis arcuatus (Pharidae: Bivalvia)

This study investigated razor clam Ensis arcuatus culture from larval rearing to commercial size to assess it's aquaculture potential. Broodstock was spawned several times in hatchery facilities to demonstrate that larval availability is not a constraint for hatchery production. Larval culture lasted 20 days, showing an average survival of 14.35%. The razor clam seed reached a size of 28.5 mm at 4 months. Seed culture is feasible; however, it is constrained by the fact that razor clams need to be buried. Higher survival was observed when culturing razor clam seed with a substrate (82–83%), irrespective of the substrate grain size, while containers held without a substrate showed a significantly lower survival (56.5%). We showed that it is possible to rear E. arcuatus juveniles in bottles, with survival as high as 94–95% being achieved. The length increase of the hatchery‐produced E. arcuatus over 600 days was described by the equation Ln L=4.5+(?141.17/t), where L is length in mm and t is time in days. On‐growing showed adequate survival rates and it took 2–3 years to reach commercial size.